Significant Association of Candidate Genes (AGTR1 and TGF-Β1) Polymorphism with Diabetic Nephropathy in Diabetes Mellitus Type 2 Patients

Abstract

Background/Aims: Diabetic nephropathy (DN) is one of the complications of diabetes mellitus (DM). This study aimed to investigate the association between genetic polymorphisms, specifically AGTR1 (rs5186) and TGF-β1 (rs1800470), and the risk of developing Diabetic nephropathy (DN) in type 2 diabetes mellitus patients, compared to those without DN and healthy controls. Methods: A case-control study was conducted on 165 diabetic patients (59 with diabetic nephropathy (DN) and 54 without DN (DM)), and 52 healthy controls (HC). The genotyping was done using amplification refractory mutation system method (ARMS-PCR). Age, gender, and duration of diabetes were matched across groups. Clinical parameters including FBS, RBS, HbA1C, creatinine, urea, SBP, DBP, total cholesterol, triglycerides, LDL, and BMI were assessed. Results: Diabetic patients with nephropathy exhibited significantly higher levels of clinical parameters compared to those without nephropathy and healthy controls. The risk allele of AGTR1, C (p <0.0001), and risk allele containing genotypes AC (p <0.0001) and CC (p-0.0010) were significantly higher in DN patients compared to DM and HC groups. Similarly, the TGF-β1 risk allele C (p-0.0001), and corresponding genotypes TC (p-0.0038) and CC (p-0.0027) were significantly associated with increased risk of diabetic nephropathy compared to DM and HC groups. Conclusion: The data showed significant association of AGTR1 (rs5186) and TGF-β1 (rs1800470) polymorphism with an increased risk of diabetic nephropathy in type 2 diabetes mellitus patients. More investigation will be required to disseminate the results, while increasing the samples size and using whole genome sequencing.

Full text

Cell Physiol Biochem 2024;58:203-211
DOI: 10.33594/000000702
Published online: 6 May 2024 203
Cellular Physiology
and Biochemistry
Cellular Physiology
and Biochemistry © 2024 The Author(s). Published by
Cell Physiol Biochem Press GmbH&Co. KG
Ihsan et al.: Genetic Polymorphisms in Diabetic Nephropathy
Original Paper
Accepted: 14 April 2024
This article is licensed under the Creative Commons Attribution 4.0 International License (CC BY). This means
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purposes, provided proper credit is given to the Authors as well as the original publisher.
DOI: 10.33594/000000702
Published online: 6 May 2024
© 2024 The Author(s)
Published by Cell Physiol Biochem
Press GmbH&Co. KG, Duesseldorf
www.cellphysiolbiochem.com
© 2024 The Author(s). Published by
Cell Physiol Biochem Press GmbH&Co. KG
Signicant Association of Candidate Genes
(AGTR1 and TGF-Β1) Polymorphism with
Diabetic Nephropathy in Diabetes Mellitus
Type 2 Patients
Madeeha Ihsana Najeeb Ullah Khana Noreen Asima Muhammad Ismailb
Mikhlid H. Almutairic Ijaz Alid Brian D. Adamse
aInstitute of Biotechnology & Genetic Engineering (Health Division), The University of Agriculture
Peshawar, Pakistan, bDepartment of Zoology, Islamia College Peshawar, cZoology Department, College
of Science, King Saud University, P.O. Box: 2455, 11451, Riyadh, Saudi Arabia, dCentre for Applied
Mathematics and Bioinformatics (CAMB), Gulf University for Science and Technology, Hawally, Kuwait,
eDepartment of RNA Sciences, The Brain Institute of America, New Haven, CT, USA
Key Words
Type 2 diabetes mellitus • Diabetic nephropathy • AGTR1 • TGF-β1 • Gene polymorphism
Abstract
Background/Aims: Diabetic nephropathy (DN) is one of the complications of diabetes
mellitus (DM). This study aimed to investigate the association between genetic polymorphisms,
specically AGTR1 (rs5186) and TGF-β1 (rs1800470), and the risk of developing Diabetic
nephropathy (DN) in type 2 diabetes mellitus patients, compared to those without DN and
healthy controls. Methods: A case-control study was conducted on 165 diabetic patients (59
with diabetic nephropathy (DN) and 54 without DN (DM)), and 52 healthy controls (HC). The
genotyping was done using amplication refractory mutation system method (ARMS-PCR).
Age, gender, and duration of diabetes were matched across groups. Clinical parameters
including FBS, RBS, HbA1C, creatinine, urea, SBP, DBP, total cholesterol, triglycerides, LDL, and
BMI were assessed. Results: Diabetic patients with nephropathy exhibited signicantly higher
levels of clinical parameters compared to those without nephropathy and healthy controls. The
risk allele of AGTR1, C (p <0.0001), and risk allele containing genotypes AC (p <0.0001) and CC
(p- 0.0010) were signicantly higher in DN patients compared to DM and HC groups. Similarly,
the TGF-β1 risk allele C (p- 0.0001), and corresponding genotypes TC (p- 0.0038) and CC (p-
0.0027) were signicantly associated with increased risk of diabetic nephropathy compared to
DM and HC groups. Conclusion: The data showed signicant association of AGTR1 (rs5186)
and TGF-β1 (rs1800470) polymorphism with an increased risk of diabetic nephropathy in type
2 diabetes mellitus patients. More investigation will be required to disseminate the results,
while increasing the samples size and using whole genome sequencing.
Dr. Najeeb Ullah Khan Institute of Biotechnology and Genetic Engineering (Health Division),
The University of Agriculture Peshawar, PO, 25130, Pakistan.
Tel.: 00923339732955, E-Mail najeebkhan@aup.eu.pk

Cell Physiol Biochem 2024;58:203-211
DOI: 10.33594/000000702
Published online: 6 May 2024 204
Cellular Physiology
and Biochemistry
Cellular Physiology
and Biochemistry © 2024 The Author(s). Published by
Cell Physiol Biochem Press GmbH&Co. KG
Ihsan et al.: Genetic Polymorphisms in Diabetic Nephropathy
Introduction
Diabetes mellitus (DM) is a complex disorder, which is very common and rapidly growing
all over the world [1]. In 2002, about 173 million cases of DM were estimated worldwide
and this number was predicted to increase by 350 million in 2030 [2]. The disease is

of the microvascular complications of DM. It is estimated that approximately 30–40% of all



hyperlipidemia, hypertension, advanced glycation products accumulation, duration of


Transforming growth factor beta () belongs to a group of multifunctional growth
factors which control various biological processes such as apoptosis, senescence, healing
of wounds, tumor metastasis and suppression, cell division, differentiation and immunity
[6]. In humans, there are three isoforms of  namely ,  and .
Among these the  is the most abundant and is highly conserved in primary sequence
 controls the production and degradation of the renal


glucose in blood activates 
and the recombinant 
in cell hypertrophy [9]. The human  gene is located on chromosome 19q13.1–13.3
             
        
codons 10 and 25 are reported to be associated with increased or decreased levels of 
production   [11]. This increase or decrease in the production of  has been
 
 gene which affect its


) is found to be a highly

        

            
          



  
   
 gene is one of the potential
        


results. Understanding the clinical implications of these genetic variants is essential for
         
with type 2 diabetes mellitus. Therefore, our study aims to investigate the presence and
association of  and       


Cell Physiol Biochem 2024;58:203-211
DOI: 10.33594/000000702
Published online: 6 May 2024 205
Cellular Physiology
and Biochemistry
Cellular Physiology
and Biochemistry © 2024 The Author(s). Published by
Cell Physiol Biochem Press GmbH&Co. KG
Ihsan et al.: Genetic Polymorphisms in Diabetic Nephropathy
Materials and Methods

This case control study was performed between February and December 2020 in the Institute of
   
     
  A written
informed consent was obtained from each participant after describing the aim of the study. T2DM was




years of diabetes duration with urinary albumin levels of  in at least two of three


     


 
protocol.

  
            


              
conditions for the  variant were; initial denaturation at 95°   
of denaturation at 95°°°
at 72°TGF
°

Table 1.




ͳ
ȋͳͳ͸͸Ȍ




 
ȋȌ
ͷǦǦ͵
ȋȌ
ͷǦǦ͵
ȋȌ
ͷǦǦ͵
ȋȌ
ͷǦǦ͵
Ǧβ1
ȋͺ͸ͻȌ

ͷǦǦ͵

ͷǦǦ͵

ͷǦǦ͵























Cell Physiol Biochem 2024;58:203-211
DOI: 10.33594/000000702
Published online: 6 May 2024 206
Cellular Physiology
and Biochemistry
Cellular Physiology
and Biochemistry © 2024 The Author(s). Published by
Cell Physiol Biochem Press GmbH&Co. KG
Ihsan et al.: Genetic Polymorphisms in Diabetic Nephropathy

           
was used for demographic data analysis with 
    

            
             
ratio calculator. The 
Results


           
               
(<0.001), Urea (
blood pressure (<0.001), Diastolic blood pressure (<0.001), Total cholesterol (<0.001),
Triglycerides ( =0.023), and BMI
(             
    
among the study groups (Table 2).
Table 2.


high density lipids, BMI - Body mass index

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ȋαͷͶȌ
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ͷ͸Ǥ͹άʹǤͺͶ
ͲǤ͸ͺͻ
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ͳͳǤͶ͹άͳǤͲͳ͹
ͳͳǤʹͷάͳǤͶͲͳ
Ǧ
ͲǤͲͺ͸
ȋȀȌ
ͳͺͲǤͲάͶͲǤͻ
ͳ͹ͺǤ͵ά͵ͷǤͲͳ
ͺ͹Ǥ͸άͺǤ͵Ͳ
δͲǤͲͲͳ
ȋȀȌ
ʹͳͻǤͲ͹ά͵ͷǤ͵
ʹͳͷǤ͹άͺǤ͸ʹ
ͳʹͳǤ͹άͻǤͲʹ
δͲǤͲͲͳ
ͳȋΨȌ
ͺǤͳ͹άͳǤͲͳ
ͺǤͳͲάͲǤ͹ͺ
͵Ǥ͹͸άͲǤ͸͵
δͲǤͲͲͳ
ȋȀȌ
͸ǤͷͺάʹǤ͵ͳ
ͳǤͲ͸άͲǤͷ͹
ͲǤͺͺάͲǤʹͲ
δͲǤͲͲͳ
ȋȀȌ
ͳͷ͵Ǥ͹ά͸͹Ǥʹ
ʹͻǤͷάͳ͵Ǥʹ
ʹͲǤͳͲάͷǤʹͻ
δͲǤͲͲͳ
ȋȀȀͳǤ͹͵;Ȍ
ͻǤʹͶά͵Ǥ͹ͺ
ͺͳǤͲͻάʹͺǤͶ
ͻ͵Ǥͳάͳ͹Ǥͻ
δͲǤͲͲͳ
ȋȌ
ͳͶͳǤͶάͳͶǤͶ͹
ͳ͵ͲǤͶάʹͷǤͳ
ͳͳ͹ǤʹάͺǤͻͺ
δͲǤͲͲͳ
ȋȌ
ͻͲǤͻάͻǤͳʹ
͹͸Ǥ͵ͻάͳͳǤͶ͸
ͺͲǤͺά͸ǤͶ͹
δͲǤͲͲͳ
ȋȀȌ
ͳͺ͵Ǥͺά͵ͺǤͻ͸
ͳ͹ͷǤ͸͵ά͵ͷǤͺ͸
ͳͶͶǤͺ͹άͳͻǤͳͳ
δͲǤͲͲͳ
ȋȀȌ
ͳͶͺǤͻάͷʹǤ͹Ͳ
ͳͷͲǤʹʹάͶͷǤͶͻ
ͳͲͳǤʹͷάͳͶǤͳ͸
δͲǤͲͲͳ
ȋȀȌ
ͻͲǤͷ͵άʹͺǤͲʹ
ͺͶǤ͵ͲάʹͷǤͺ͹
͹͸Ǥʹ͹άͳ͵ǤͲʹ
ͲǤͲͲ͹
ȋȀȌ
͹͸ǤͲʹ
͹ͺǤ͸͹άʹʹǤ͹ʹ
ͺͷǤͺͺάͳͷǤͲ͸
ͲǤͲͷͶ

͸͵ȋ͵ͷǤͷȌ
ʹ͹ȋͳ͸Ǥ͹Ȍ
Ǧ
δͲǤͲͲͳ
ȋȀʹȌ
ʹͶǤ͵ͷάʹǤͺ͵
ʹͷǤ͵ͷάͳǤͷ͹
ʹͷǤͶͷάͲǤ͸͹
ͲǤͲͳͳ
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Cell Physiol Biochem 2024;58:203-211
DOI: 10.33594/000000702
Published online: 6 May 2024 207
Cellular Physiology
and Biochemistry
Cellular Physiology
and Biochemistry © 2024 The Author(s). Published by
Cell Physiol Biochem Press GmbH&Co. KG
Ihsan et al.: Genetic Polymorphisms in Diabetic Nephropathy


The allelic and genotypic frequencies of 
    
              
            
     =<0.0001,
=0.2055,


            
   
=0.2903) (Table 3).
Fig. 1.          
   
   
  


Cell Physiol Biochem 2024;58:203-211
DOI: 10.33594/000000702
Published online: 6 May 2024 208
Cellular Physiology
and Biochemistry
Cellular Physiology
and Biochemistry © 2024 The Author(s). Published by
Cell Physiol Biochem Press GmbH&Co. KG
Ihsan et al.: Genetic Polymorphisms in Diabetic Nephropathy
Discussion
 is said to be an important regulator in the production and degradation of the


the renal hypertrophy could occur because of higher concentration of , such changes
 
          
   
activation, increased intraglomerular pressure and hypertension, which are said to induce
the production of 
 affect the
 is located
on the chromosome 19 (q13.1-13.3) and has a total of 7 exons and 6 introns [23]. Various

      

 

    


              
mellitus type 2 for over 10 years. Their result suggested that the  gene polymorphism
is associated with susceptibility to diabetic nephropathy [22]. Another similar study was
carried out in patients with type 1 diabetes to see if it contributes to genetic predisposition
   
               
TT genotypes may be protective factors [3]. These results were in accordance with our
           
   
 gene polymorphism with diabetic nephropathy. Their results suggested that 
gene polymorphism is not associated with the progression of diabetic nephropathy [25]. A

Table 3.

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ȋαͷͻȌ

ȋαͷʹȌ

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χʹ

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ȋͳͳ͸͸Ȍ
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ʹͲȋ͵͹ǤͲȌ
ͳʹȋʹ͵ǤͲ͸
Ȍ
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ͻǤ͹͸ȋ͵ǤͺͻǦ
ʹͶǤͶͺȌ
δͲǤͲͲͲͳ
ͳǤͻ͹ȋͲǤͺͶǦͶǤ͸͸Ȍ
ͲǤͳͳͺʹ
ͶǤͻ͵ȋʹǤͲͻǦ
ͳͳǤ͸͵Ȍ
ͲǤͲͲͲ͵


ͳͲȋͳ͸ǤͻȌ
Ͳʹȋ͵Ǥ͹ͲȌ
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
ͳͷǤͺ͵ȋ͵ǤͲ͵Ǧ
ͺʹǤͷȌ
ͲǤͲͲͳͲ
ͳǤͳͺȋͲǤͳͷǦͺǤͻͳȌ
ͲǤͺ͸͹͵
ͳ͵Ǥ͵͵ȋʹǤͷͶǦ
͸ͻǤͻͲȌ
ͲǤͲͲʹʹ

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
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ͲǤͳͶͷ͵
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ͲǤͺͶͶ
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ͲǤ͹͵ͻ
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͸ͳȋͷͳǤ͸Ȍ
ͺͶȋ͹͹Ǥ͹Ȍ
ͺͺ
ȋͺͶǤ͸ʹȌ
δͲǤͲͲͳ



ͷ͹ȋͶͺǤ͵Ȍ
ʹͶȋʹʹǤ͵Ȍ
ͳ͸
ȋͳͷǤ͵ͺȌ

ͷǤͳ͵ȋʹǤ͹ͲǦ
ͻǤ͹ͺȌ
δͲǤͲͲͲͳ
ͳǤͷ͹ȋͲǤ͹ͺǦ͵Ǥͳ͸Ȍ
ͲǤʹͲͷͷ
͵Ǥʹ͹ȋͳǤͺ͵ǦͷǤͺͶȌ
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Ǧβ1
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ȋͺ͸ͻȌ
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ͲǤͳͶͳͷ
ͳǤͻͺȋͳǤͳ͸Ǧ
͵Ǥ͵ͻȌ
ͲǤͲͳʹ͵

Cell Physiol Biochem 2024;58:203-211
DOI: 10.33594/000000702
Published online: 6 May 2024 209
Cellular Physiology
and Biochemistry
Cellular Physiology
and Biochemistry © 2024 The Author(s). Published by
Cell Physiol Biochem Press GmbH&Co. KG
Ihsan et al.: Genetic Polymorphisms in Diabetic Nephropathy
   
            

 gene is
           

            


      gene polymorphism with susceptibility to




 
to patients with AA genotype [27]. Another study conducted and observed the association
of the 

 
 genotypes between the patients and the
  
 
    
       

 gene polymorphism

Conclusion
In conclusion, our research suggested an association of allelic and genotypic frequencies
         
        

while using modern sequencing technologies.
Acknowledgements
 


There is no funding received to conduct the study, the authors contributed for the study







study.

Cell Physiol Biochem 2024;58:203-211
DOI: 10.33594/000000702
Published online: 6 May 2024 210
Cellular Physiology
and Biochemistry
Cellular Physiology
and Biochemistry © 2024 The Author(s). Published by
Cell Physiol Biochem Press GmbH&Co. KG
Ihsan et al.: Genetic Polymorphisms in Diabetic Nephropathy



All the authors have read and approved the article for publication

All the necessary data are included in manuscript, related data will be provided on
request from corresponding author
Disclosure Statement
The authors declared no competing interests
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

 


 

 

2016;45:761–767.
 

 
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 


 

 

 
transforming growth factor-beta1 gene: association with transforming growth factor-beta1 production,

 

 

 
contributor to dyslipidemias, altered glucose homeostasis, and hypertension of the metabolic syndrome.


Cell Physiol Biochem 2024;58:203-211
DOI: 10.33594/000000702
Published online: 6 May 2024 211
Cellular Physiology
and Biochemistry
Cellular Physiology
and Biochemistry © 2024 The Author(s). Published by
Cell Physiol Biochem Press GmbH&Co. KG
Ihsan et al.: Genetic Polymorphisms in Diabetic Nephropathy
 

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
2010;37:152–157.
 

 

 


 
 

 


 

2004;13:1649–1656.
 

 


 


 


 󰨠

 

 
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