Experiment FindingsPDF Available

Genetic Association of Angiotensinogen (M235T) Gene Polymorphism with Essential Hypertension in Vindhyan Population of Madhya Pradesh, India

Authors:
  • Centre for Rural Development Ecology and Environment Protection (CRDEEP)

Abstract

Essential hypertension is common in the general population by its increasing incidences, thus recently researchers suggested that AGT (M235T) polymorphism may be the functional genotype in hypertensive patients. Therefore the study aimed to evaluate Angiotensinogen (AGT) gene polymorphisms (M235T) as well as its relation to serum level of renin and aldosterone among Sudanese hypertensive patients. The study concluded that there is no association between the polymorphism (M235T) of the AGT gene and essential hypertension, whereas association observed with serum renin and aldosterone levels. The descriptive data and comparison of biochemical parameters of essential hypertension patients versus healthy controls are presented in Table no. 4.2. As expected the essential hypertension patients had markedly higher levels of HDL-C (P<0.0001), LDL-C (P<0.0001), TG (P<0.0001), Systolic BP (P<0.0001) and Diastolic BP (P<0.0001) compared to that of control subject. Significant level of change has been seen in overall distribution of Angiotensinogen (M235T) genotypes in Healthy control group as compared to disease group although Healthy control group showed little increase in ‘MM’ genotype as compared to Patients of Essential hypertension (69.44% vs 56.25%). Similarly, mutant type ‘TT’ genotype was present in low frequency in Essential hypertension patients group 3.75% and also in control group 4.44% (χ2 = 0.0243*, P= 7.436).


       
!"#$$$%&$'
()*+%,-.)/$,)..*
1Centre for Biotechnology Studies, APS University, Rewa, Madhya Pradesh, India.
2Department of Zoology, CGPG College, Panna Madhya Pradesh, India.
A R T I C L E D E T A I L S A B S T R A C T
*' &$
    !  !" #    $   ! %!
& #  !   "  "   ' 
 % " #""   !  ! ( AGT )*+
,--!"'.+ /! 
 "!"#!%%!#'0.+!!
# ! %! '.+"  ! 
)*+!)!#+! !*!!1!.+'
!.+)*+ !  #!&!#.+&%!!
1!!)*+%!! .+'. !  
!.+)*+#%!!!!'+!
"  *+  !  2            %!
!'!)*+   !.+!!
!'32 #     )*+ !#!%"#%
!%!!'
*4!#(Centre for Biotechnology Studies, APS University, Rewa, Madhya Pradesh, India.
Received: 15-Feb-2024; Sent for Review on: 24-Feb-2024; Draft sent to Author for corrections: 10-March-2024; Accepted on: 18-March-2024
Online Available from 20-March-2024
56(7
8© 2024 CRDEEP Journals. All Rights Reserved.
0.,10-!!2
21133 '$4'33434,35
&26$$)
&))(1778*9*)2&1:'*;;

<0.$8. $=2
Corresponding Author:
9 
Key words:
.: ! &
:&.+&
;!&
<5<&=5<'
; !  ! #  &!
!!.+)*+ !  #!
!"'+!!  "  .+ 
 ! )*+% " 
!"'+! !! #%!
 ! )*+  ! .+    !& %! 
#" %!     "' +! "     
#!  !"!!
+#'''1!!! $!!"
HDL-C :>'& LDL-C :>'& TG :>'& Systolic BP :>'  Diastolic BP
:>'  !   #/' "  ! ! #  
" #   )*+   =!   
     !! =!   !%    ?))@
    : ; ! ,-'A vs *,'*A'  &
  ?++@  %   % 2  ; ! 
'B*A'Aχ2 C'4&:CB','
Soni et. al.,International Journal of Basic and Applied Sciences 13 (2) (2024) 10-20
+!#%  ! 9:!.+59%"!
' +! ,--1 +D 1! ,E   ! )
!+ ),E +'+!,--%"   *&& !&  )*+' 
#% & !  !  .+ ,-- ! #   #  %! " # 
! &*'!9:!.+&B,&D+#1!.+B'
!&!1!!+ !)&$%+B)+B)&'+!
B,   $  !  # " 
!,&E')"&!$!%#%!
B,,--!'6"!%&!! !
" !! !!%'+!"%1
 !!"! !
!'+!.+)*+ ! 9: !)!+! 
#*&!)+&"'=%"&#!!"#,'
+!!&%!!#" %! 
!)*+ ! !.+$!*-'
' $$1
2.1 Study population:
+!       #/    , ; =   E
! !%!'!=&)  
$! "# @#=!'
2.2 Inclusion and Exclusion criteria for Cases:
D "  %!  ! 5   )& ! !! ) D  /
.! )  =& %& " ) D&%& #1!    %&
5!&!!&!'=%%!F=!6GF=6
;1 '
2.3 Inclusion and Exclusion criteria for Controls:
D     # !! " ! %   H5# % D I
G#)D% "% '+!
#/% !! ! 0!%!
!'. 0D E
'>$61
2.4.1 Anthropometry:
=!F!% !!%!!'
)1 ) %   %!  $  " # !!    2' F   %
 %#%% ! % 
1  %!F=%%! '#
%  %  ! !          *   
!   !"!%%'
2.4.2 Biochemical Analysis:
!    #%  #! #/' ) 
 "+!+D&++.&=#&=!!=5<D&<%
!<5<D  J%  #  !  !    
! GD#!5&)! &. '
2.4.3 Blood collection and plasma/serum separation:
0# %# !#/!"!"%!%!
' &   !""%#
!#  'KD
',1
2.5.1 Method for DNA isolation:
11
Soni et. al.,International Journal of Basic and Applied Sciences 13 (2) (2024) 10-20
. 59%1 %!##! ##)%$
)et al'-EE'3G# %!%  ''* '%!# %
' '#')& ))& )+A+L'* ' #'+!
1% 1#"!#% '+! 1%!% '
  '+! 1%& * 'KD!
' +!%  / &    !!
# #'+!%' '#& * '+!
% ! "  ' '  G " %  1 !!  "1' +! 1 %
&  'KD'%'+!#"&E 'μ
M#'B*)9&');5+&=E' 'A5A%8"5&=B'%')1μ
%%!%!!! %'% & 'μ
*)9%!$"*'+!#" 1' 'G&"
%' ' !! ( "8"%    "   !      '
)#%"%! $':!%#!
#" 1&  ' KD'2%"! 
#'+%"'+!2& '!#!%!
#%"" !59'59$!'+!%&
 'KD!59!'%'+!%%!%%! 'BA!'
+! 1%&  'KD'%%
'+!59%! '+;#=B' )+=D<=B'& );5+&=E''59μ
%%""!BKD#2G'
2.5.2 Determination of quality and quantity of isolated DNA:
+!59#:D#'+!#:D%!G! 
!    '  59  %2
2 &#!%!'
2.5.3 Quantitation by UV spectrophotometry:
+! 59%!# !##" &, &E 
 J0"#! &'59%#!,
 8E 1 'E %''+!##, %!
 59&!!!#59* 8 !##C', 'μ
2.5.4 Agarose Gel Electrophoresis:
. !  !  59%   2"      '  59
#' !G #  !   .& &
%'#&* ! 59%'E'EA%8"& ! μ
# '* 8 !%E01+;# )+& )& )μ
;5+'< #59Eco 8ІHind #.&&% %! $Ш
 !&!59#%"G!!J0  
  0#< &D1&3"'
2.5.5 Polymorphism screening1
&! 591 !#!!""%#/
:D%#!#!!"'!
:D%:+D! )!'J':D&1"  
" #59  !  59 &"%!!
59"2'5!1 "&%$ !
 1%!! 59&%!!"% '"
%!:D!& ! :D 1""
'+!":D%!:D 1159%!!%#=:<D
 %   9& 59&       !   # !  2'
#2G % ##!#59%! 
G &"# #!  !
G 'G   59'6G 
! G    #59!!
 ' +!#! " ! G  !! !!#$# 
12
Soni et. al.,International Journal of Basic and Applied Sciences 13 (2) (2024) 10-20
592':%$""G 'F!59%!
 G    & "       !   
2'!#H"I%!!#!!=NE'%!&1 
!!G NJ8 59"!''!&5)6'+!μ
:D% G" !#'+!
:D%!%! !
G'.%%!! # '* 8 #2"G!!J0μ
 '
':-)($1
: ! +!.+)*+ ! !9:,--!'+!
.+)*+ ! 9: !)!+! #
*&!)+&"'
2.6.1 Primer sequence:+!2 %!##))! (Shamaa MM et.
al. 2013).
3% (*ODD.+++.+.D...DD+..D+D+D+O
": (*OD...+.D+.+DDDD+..DDDDO
2.6.2 PCR Mix:
+!:D%  "   * &  *     59* & '*   L μ μ μ Taq
 # )+=D=E'E&* )MD&'* ))D&'A&'*A+%&'*A9:P
LP.1!:"'<'&&  )9+:.&&& *μ μ
 8 %"  J8 +259 'μ μ μ
2.6.3 PCR Thermal Program:
* -*KD&! %#/*-*KD &*EKD*&
BKD*&%!1 BKD! '#%! !#
'*A!'
2.6.4 Restriction digestion By PsyI:
:D    )*+ . %!   " ,*#   %!! %
%! PsyI G  9% ; #& "& )  , ! BKD' +! %  )) % 
&%!! ! G++%###'+!!G
+D% ,*##%!#59  !
#! !%!#!'*A'+!
% # !"#  '
'7)1
%# !#2&2
!   !     ' 5   5#  %!  
!!=D!'+!
$%2'+! %#!! #"!"
!! # "!  '+!  
$%2'+! %#! #
!"!! #'+!%! #
"!"#! #"'5%G
);1&)D'6!#!  &#%##
!##%!@' !#!  #%
! %! %!#%'+!:"
%!! 2%'% $&
: '&: *'"'
' 
3.1 Results:
+!" ! #!  !"
+#'''+!&1&)&F=%! '1!!
13
Soni et. al.,International Journal of Basic and Applied Sciences 13 (2) (2024) 10-20
! $!!"%! :C'-,!% :C'B)F :C'B
):C'-- # #! %    #%   !! ' +! F=  F 
:C'B):C'-B%#%'
,*'0$
0 0*:? 0*;? 8.
3@ ,8*, EE8,
 *'*Q'* *',Q' '-&
" ,'*Q' ,'Q' '*&
@A
@ ,'*Q'B ,',Q'* 'B&
 ,E'Q*', ,B'EQB' '-,&
6&/3
@ *',Q' ,'Q' 'B&
 ',Q'B *'Q*' '--&
@
@ -'*Q,' -',Q,'B 'E,&
 -'QB' E-'Q,' '*B&
"
@ -*'-Q' -,'Q' '&
 -'EQ' -'Q'* ',&
@"
@ '-BQ'* '-EQ'E 'B&
 '-EQ'E '--Q' '-B&
4"!#%
3.2 Biochemical and clinical findings:
!  !# % !%#'
%#@"!"%!!' +!"
 #!  !"!!
+# ' ''  1 ! !  ! $!! "  HDL-C :>'& LDL-C
:>'&TG :>'&Systolic BP :>' Diastolic BP :>' !#/'F!"&
 %#%!!'
,'Comparison of Biochemical & clinical findings of essential hypertension patients and healthy controls.
0 0*:? 0*;? 8.
8$3- E'BQ' -'Q', '*-&
",*0B *'-Q'B *'EQ'E '*&
"-803 'EQ' -'Q', :>'444
-803$ -'Q', 'EQ'B :>'444
3$ '-Q' ,'Q' :>'444
)6" *'QE' 'EQ*'B :>'444
-6" -B'Q*'E E,'*Q,' :>'444
6$<3$ ,',EQ'B ,'EQ'E '*-&
<03> '*EQ'-, ',Q'*B ',,*&
)3$ -'Q' -',Q'E '-BE&
<3> ,'Q'B ,'-Q' 'B*&
<3> B'*Q' ,'EQ' 'B&
(* denotes the level of significant change between case and control)
3.3 Detection of Genetic Polymorphism in Angiotensinogen (M235T) Gene:
:D %! ",*#%!!%%! PsyI G 9%;#&
"&),!BKD'+!%))%&%!! ! G
++%###'+!!G+D% ,*#
#%!#59  !#! '
14
Soni et. al.,International Journal of Basic and Applied Sciences 13 (2) (2024) 10-20
8*1")*+ ! '<)*#  $&
<))F&<)+!G<++"'
+!#! ! )*+%%!=F#2# =F;
!!'+!#"2&2)*+
 ! #'*#',.!&&'"!!#"
#)*+=! !!=!
!%?))@ :;!,-'Avs *,'*A'
 & ?++@%%2;!'B*A
'Aχ2 C'4&:CB','?++@ !! #"
#%#!'
))'*,*B%!!"%!)+'EEB
;!"! %!
!;!#'6"?)@%%2 
=D %!  ?+@ %    !! 2  !   # !     
χC'B4&:C',-'D?)@%!!!! 
!!B*'BBA0,E'B*A%!?+@%1  #!
"!!#'+!#&2
      )*+  !    %! ;
!'
,'32#.&2)*+
 ! 0!D!2+

!(!
0)!
(C*:?
(B
0D(D
(C*;?
(B
0"&)=<!#<!
χ#




-*,'*
,'
,'B*
*,-'
B,'
E'
B','4

B,'*
B,'B*
-BE'*
,B'*
',-'B4
0
*,E'B*
B'*
BB*'BB
**'
'BB'-*E
(* - denotes the level of significant association between case and control.)
(N – Number of individuals in study group.)
(% - Genotype allele frequency and carriage rate expressed in percentage.)
15
Soni et. al.,International Journal of Basic and Applied Sciences 13 (2) (2024) 10-20
,>'3!;1+")*+ !


!(!
0)!
(C*:?
(B
0D(D
(C*;?
(B
# D$$
9B$.




-*,'*
,'
,'B*
*,-'
B,'
E'
'4
'BB44
'B-
'*,*B','EE
'EEB'-'-E,
'EBB'E',-

B,'*
B,'B*
-BE'*
,B'*
'*-4
',E',E'--
',E''*
0
*,E'B*
B'*
BB*'BB
**'
'
'B*',,',*
''-EE'*
(* - denotes the level of significant association between case and control.)
(N – Number of individuals in study group.)
(%-Genotype allele frequency and carriage rate expressed in percentage.)
('8*1.32)*+'
>' -1
+!# =+9!!!/# #!!!!!!
!'J# %: !"    "  %!&
:&=+9'!!$#!!!!%! 
:&"!!:!%$%#'%#%!
 !   ! !
#"!&  B&'+! !" & /
 !&"!# %&! !  &
%! !  ! " "   !    !! B !α
'+! #$ !! !"!!!
" !! #G&  !
' M # #  ! ! # % 
 $ !    ' =     #   ! " 
"#       !'  $
!!"&"#'
16
Soni et. al.,International Journal of Basic and Applied Sciences 13 (2) (2024) 10-20
('8132)*+'
=&!$"&  #&" &
 ' ;"    "%!8#& !! & 1"   & 2
 &!"& !*':"!
#!"!!8#'3!&!""
"   %&  &    "' +! D! D )& #"
! !&"&!! & "!!'
6 G!"&&!2 ! #!
$%:B';!! #$#!
  ')$    "  !P! #2
 ""# #" !'F"%
!%!!!!" #!!
!!"#GE')!!!)!% ' ") %
!!%!"! "  %! ! "')/
 !" %! /#%
1')!!!$   % %!!!
 %!$   '  %!      
#!!%!!*-'
6!!!! $!!"%! :C'-,!
% :C'B  )F :C'B ) :C'--##!%  #%
!!'+!F=F :C'B):C'-B%
#%'!  !# % !
%#'%#@ " !" 
%!!'+!" #!  !"
!!+#''1!!! $!!"
HDL-C:>'&LDL-C :>'&TG :>'&Systolic BP :>' Diastolic BP :>' !
#/'F!"& %#%!!'
+!)*+ ! !  .+"'+!!#%.+
 ! D5!# "   % & !%"& %!  '    !
&%! !1'!
&%!!+%#/%!%!'+!*B,
%!%!&,%!D5!D5%!D5'
+!!#$!" !&)*+"!&
%! "&  !"! "
"'+!%1 %!!!)*+"!
#$!"   !     ' : 
!""%##"%"% !'
% #  !!2'
17
Soni et. al.,International Journal of Basic and Applied Sciences 13 (2) (2024) 10-20
F!!%BEA&!%#%!
!"!'3! &!%!
#% ""  !  ! ' +!     !
!!! ! )*+"  !    $  ! "    
!'
;! && 1 "%!!!
%!" !#&$&! ' ! 
!% "& !!#!
! $!  *'+!1 !% ! !8
85D;)*+.+"!& #! 
"'+!85 ! !D;)*+ ! !.+% #3<:
 ! ! ,E!"B "
#,'"!% #! #!"'9%
#"!2#D; ! #%!%&%!
.+++! G% 2',A"''BA&:C'E'+!# C'*% 
B'
6 ! )*+"!!:D %! ",*#
%!!%%!PsyIG 9%;#&"&),!BKD'+!%
)) % &%! ! ! G ++% # #  #'+!
!G+D% ,*##%!#59 
 !#! '+!#! ! )*+%
 %! = F# 2# =F;  !! ' +! #"  2& 
2)*+ ! #'*#',.!
& & '  "  ! ! #   " #   )*+   =!
       !! =!   !%    ?))@  
 :;!,-'Avs *,'*A' & ?++@%%
2  ; !   'B*A      'A χ2 C '4& :C B',' ?++@
!! #"#%
#! '     ))   '*,*B %!!   "  %!     )+
 'EEB ;! "         !
%!!;!#'6"?)@%%2
 =D%!?+@ %!!2!#!
 χC'B4&:C',-'D?)@%!!!!
 !!B*'BBA0,E'B*A%!?+@%1  #!
"!!#'+!#&2
)*+ ! %!
;!' +!)*+ ! !!!"
! !!!!$'+! !%!
#%)*+   !  ! $  "!',
  *  %! !  *   1 ! !!  %     
  !2' D#! "#% ! $ '
  ! !%1#'+!.+)*+%
 !G#!'<%!-*A
"D% !$.+)*+ ! %!!'
+!    !  $   !& ! &  !& & % 
!&!!!"&%!!%!! 
:>''+!.+)*+!++!+%!$
! !;!'# #!
#%.+=+9' . ! $!R.+ %#
%!#!!59!*'
+!!!;=#" ')!
#:"%##!!'+!!
  !"#1"#  !%  !  !
18
Soni et. al.,International Journal of Basic and Applied Sciences 13 (2) (2024) 10-20
 :'  ! # " !% !  .+   !  #  %!
$;=B'+!"!2.+)*+ ! ;=
;' ! ! : !3<::D%
 ! )*+ !  .+ ! #   ,' +!
 "! ! % " $  " ;= %! !"! +  %!!  ! G 
!G'%!!%#%.+)*+ ! !$
";=,E'
' 01
+! ! !!:# "
  ! # %  '  .+   !   !  ""  !
"8   ! ' .  !      !   .+ )*+ 
#%!!!;='"!!#"#
)*+=! !!=!
!%  ?))@   : ;! ,-'A vs *,'*A' &
?++@%%2;!'B*A
'Aχ2 C'4&:CB','?++@ !! #"
#%#!' 3&% G& #!  
1!!! $!!" =5<D&<5<D&+.& :5:
 !#/%%!;!'=& "
)*+%%!;!'
:' 1
' ;.%& ')'  " ; D # D ;1  D .1 =  <
$'''''*&&R'
' <"&;'P!&'P3&')'P3&3''P3&:'. ! ! 
'M'&,&,R*'
' :&:'P+%&'M'PM &M')':'P &'D'P.&'P.&'P! &'M'P&''P9&'PD!&+''P
'D! %!#(' ! ')D
)'.',&B&'
' )& .'P D!"G& '6'P.& 'P :& <'P +!& 5'P & )''P0& <'P 3& 3' +! D$ #%
        ;  . )#  5#
:"'D'0':! 'E&,&R'
*' !&''P9&'.'P+#/S=&'.: ! &: &$
=! =5''+! #'0''&&,-RB*'
,' D!& =''P D!& D'='P !& M'='P D!& D'='P <& '5'P F& )'T'   !  !   &
"G  #%!  +%''
D!')''&,,&*R*,'
B'  &'''P. &;'='P=&U'+'P&.':" !% ;
(+!*;=''= '='&RE'
E' & M''P ;#& 3''P M& ;'P 5 & ='P ;& T'P .G& .'P 5& J''P & )'P V G& 6'P W& +'P  '
!)*+:6;.: ! +$!+5#:%!
%!9!!''3'&&,-RB'
-' =&='5'P<&3''P<&L''PF&<''P&.''. ! ! 
D!%!!!'D!')''&&ER'
' #&'<'P<&='T')!  DM5'' ''9!'&&,R,'
' & 'T'P D!& 'D'P T& '='P M & F'T'     !# F!
D"5 - D605-6 M( 9% :#D!'D'
'5'&B&RE'
' <X"2& 'P )2&')'P <& D'P & )'D'P& 3'    .+ =  )
F!:='=&&BRBE'
' <& ''P )& 'T'P <& '='P ! & D'.'P <& +'F'P M & 'M'P D!& '='P M& 'F'P M & +'='P :$& ''P  '
: ! :+5#)M+#/'M
:''&B&-*R'
'  !& <''P DY.&)'P D& 'D'P)1%& ':'P )M!& '' .  #%   !
 ( "% ')6P.&
&-
19
Soni et. al.,International Journal of Basic and Applied Sciences 13 (2) (2024) 10-20
*' D!!&5'P+&)'P<"&)'5#!!Z %!'5"')'&&
,R*'
,' !%&+''P!/&)'P&+''PM!&=''P!&'P&M'PM!&)'D;0%!!:!%
D$:+5#9!!'59D'-&E&R*'
B' )$&'P[&)'PU"\$&)'PD\$&'P:"]&5'.+  ! ,--
B,#!!D%!# ''')''B&B&,R
,B'
E' %&J'D'P:G&)''+! !$(;$' ''
)'&,&,RB'
-' . !& 'P U! & )''P .GGG$& F'P U$%$GG!%$& ;'P ! ; 5  .'
"G  8)*+  ! ($!
'M'&*E&*R*-'
' :$& :'P !& :'D'P :!& 0'M'P 9& ')'P $& '9'   .
: !  + 5#:  !)%: /!'' '')' '&&
ER'
' 5!&+'P^&^';5D!M5;M3'
:'B&&B-REE'
' ;9#&'='P&'P#;!G&)''P;3$&)'P) &';''P;.%&'B*.: ! !+
<$  . %!D " ; )%';&
)#' 5'5+&&,-R,'
' ;.%&')'P=#;9#&';'"   5 J 5 )! 
6"35.;!)
+'D'':'',&&E,*REB'
' .! &+''P3&';'P &''P;6$&'P;=&T'; $!$
!;!$."&;''M5'+',&B&'
*' ;.%& 'P ;9#& '='P .!& )'P # & ;'  #% 39_ . : !  
6 + %!5"D!=D0;:' '"'&
*&R'
,' !$!&'P!!&')'P)&^'P &)'PG!&'."D;85.+),E+
%!#!!'''&*&R'
B' U!&+''PT&''P^&T'=')*+ ! %!
$( ')'''&&B,*RBB'
E' +$$& T'P T!& +'P T!$& M'P J $%& +'P M& 'P +& ='P M%& M'P 3$& 'P T!$%& +'
 ! )*+!"##
% ')# ,&**&E-RE'
20
ResearchGate has not been able to resolve any citations for this publication.
ResearchGate has not been able to resolve any references for this publication.