Investigation of the antidiarrhoeal activity of Holarrhena antidysenterica

ArticleinIndian Journal of Pharmaceutical Sciences 64(2):164-167 · January 2002with 44 Reads
Abstract
Holarrhena antidysenterica (L)-Apocyanaceae, well known for its antidiarrhoeal activity was studied for its effect on diarrhoeagenic Escherichia coli. Different dilutions of the decoction of the plant were assayed for its effect on the adherence and toxin production of 2 groups of E.coli- enteropathogenic (EPEC) and enterotoxigenic (ETEC.) Adherence per se was not affected though disruption of the characteristic 'microcolonies' of EPEC on HEp-2 cell line was observed. The decoction was more effective in inhibiting stable toxin production as compared with labile toxin production.

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  • ... HA inhibits the stable toxin production and prevents its intestinal secretions, which leads to a decrease in the virulence of enterotoxigenic (ETEC) strains. Thus, it can be concluded that HA gives protection against multiple stages of diarrhea.[21]Srivastava and Saxena studied the in vitro activity of the aqueous extract of HA seeds against E. coli, Shigella, Staphylococcus aureus, and Salmonella typhi organisms and found it highly effective against these pathogens responsible for diarrhea.[22]In ...
    Article
    Holarrhena antidysenterica (L.) Wall. ex A. DC. is a medicinal plant abundantly found in India. Its uses are mentioned in the classical Ayurvedic literature and by many folklore claims. The plant is also of extreme economic importance. Its seeds are mainly used as an antidiabetic remedy. All pharmacological and toxicological aspects of this plant are discussed in this review.
  • Article
    Holarrhena antidysenterica (L) (Kutaja-Wall) Apocynaceae, is a common plant widely cited in literature for its medicinal value due to the presence of a large number of alkaloids. Various parts of the tree viz. bark, root, stem and seeds are known to have various medicinal properties including acute and chronic diarrhea and dysentery activity. The alkaloids are present in the polymeric matrix network of proteins and polysaccharide. There are specific enzymes that can break down this proteinaceous and polysaccharide matrix leading to an increase in the release of the drug, which will then facilitate the extraction. The overall effect will be an increase in the overall extraction of the constituents of interest. It was observed that cellulase was most effective in the release of alkaloids.
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    Ethno-medico-botanical knowledge was documented in Hosanagara taluk in Shimoga districtof Karnataka, India using a questionnaire. The data of 86 plant species for treating 47 human and 16 veterinary ailments were analyzed by informant consensus factor (ICF), use value (UV), and fidelity level (FL). The wound category received high ICF of 0.86. Vallaris solanacea (UV=0.61) for bone fracture and wound and Tabernaemontana alternifolia (UV=0.54) for snakebite were preferred by healers. The FLs for a specific disease was maximum (100%) for Aristolochia indica, Calycopteris floribunda, Thottea siliquosa, Tridax procumbens, and Tabernaemontana divaricata. In this study, 14 hitherto unknown ethnomedicinal formulations are reported.
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    Zingiber officinale (ginger) was studied for its antimicrobial profile and effect on virulent features of diarrhoeal pathogens, viz. colonization of epithelial cells and production of enterotoxins. Z. officinale showed no antimicrobial activity. Although it inhibited the production of cholera toxin, it had no effect on the action of this toxin. It also had no effect on the production and action of E. coli heat labile and heat stable toxins. However the bacterial colonization of HEp-2 cells was reduced. The results indicate that in the absence of antimicrobial action, Z. officinale exhibits its antidiarrhoeal activity by affecting bacterial and host cell metabolism. The present study reports a novel mechanism of action by Z. officinale in infectious diarrhoea.
  • ... issoo leaves also highlights the necessity of including multiple parameters for judging the efficacy of medicinal plants. Assaying bacterial virulent features as a marker for demonstrating the antidiarrhoeal efficacy of a plant, has been previously reported by us using two indigenous plants viz. Cyperus rotundus [17] and Holarrhena antidysenterica. [18] This is especially important in the absence of antimicrobial activity, which in most of the studies reported earlier [19][22] has been considered the marker for antidiarrhoeal activity. ...
    Article
    Full-text available
    Objective: Several medicinal plants have been evaluated for their antidiarrhoeal activity. Most studies evaluated their effect on intestinal motility and antimicrobial activity and, therefore, did not take into account the pathogenesis of infectious diarrhoea. Features of b Naoroji Godrej Centre for infectious diarrhoea like abdominal pain, cramps, inflammation, and passage of blood/mucus in the stools are the combined effect of one or more virulence factors of the infecting organism. The effect of medicinal plants on the microbial virulent features can serve as marker(s) for testing their efficacy. In this study, we evaluated the effect of a decoction of dried leaves of Dalbergia sissoo on aspects of pathogenicity, that is, colonisation to intestinal epithelial cells and production/action of enterotoxins. This was done to define its possible mechanism(s) of action in infectious diarrhoea. Material and Methods: Antibacterial, antiprotozoal, and antiviral activities of the plant decoction were checked by agar dilution method, tube dilution method, and neutral red uptake assay, respectively. Cholera toxin (CT) and Escherichia coli labile toxin (LT) were assayed by ganglioside monosialic acid receptor ELISA. Suckling mouse assay was used to assess E. coli stable toxin (ST). As a measure of colonisation, the effect against adherence of E. coli and invasion of E. coli and Shigella flexneri to HEp-2 cells were studied. Result: The decoction had no antibacterial, antiprotozoal, and antiviral activity. It reduced the production and the binding of CT and bacterial adherence and invasion. Conclusion: This study showed that D. sissoo is antidiarrhoeal as it affects bacterial virulence. However, it has no antimicrobial activity.
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    Psidium guajava L., Myrtaceae, is used widely in traditional medicine for the treatment of diarrhoea, dysentery, gastroenteritis, stomachaches, and indigestion. However, the effect of the leaf extract of P. guajava on the pathogenesis of infectious diarrhoea has not been studied. The present study evaluates the effect of a hot aqueous extract (decoction) of dried leaves of P. guajava on parameters associated with pathogenicity of infectious diarrhoea. The aim was to understand its possible mechanism(s) of action in controlling infectious diarrhoea and compare it with quercetin, one of the most reported active constituents of P. guajava with antidiarrhoeal activity. The crude decoction and quercetin were studied for their antibacterial activity and effect on virulence features of common diarrhoeal pathogens viz. colonization of epithelial cells and production and action of enterotoxins. Colonization as measured by adherence of enteropathogenic Escherichia coli (EPEC) and invasion of enteroinvasive E. coli (EIEC) and Shigella flexneri was assessed using HEp-2 cell line. The production of E. coli heat labile toxin (LT) and cholera toxin (CT) and their binding to ganglioside monosialic acid (GM1) were studied by GM1-ELISA whereas the production and action of E. coli heat stable toxin (ST) was assessed by suckling mouse assay. The decoction of P. guajava showed antibacterial activity towards S. flexneri and Vibrio cholerae. It decreased production of both LT and CT and their binding to GM1. However, it had no effect on production and action of ST. The decoction also inhibited the adherence of EPEC and invasion by both EIEC and S. flexneri to HEp-2 cells. Quercetin, on the other hand, had no antibacterial activity at the concentrations used nor did it affect any of the enterotoxins. Although it did not affect adherence of EPEC, it inhibited the invasion of both EIEC and S. flexneri to HEp-2 cells. Collectively, the results indicate that the decoction of P. guajava leaves is an effective antidiarrhoeal agent and that the entire spectrum of its antidiarrhoeal activity is not due to quercetin alone.
  • ... Cyperus rotundus [17] and Holarrhena antidysenterica. [18] This is especially important in the absence of antimicrobial activity, which in most of the studies reported earlier [19]- [22] has been considered the marker for antidiarrhoeal activity. ...
    Article
    Full-text available
    Objective : Several medicinal plants have been evaluated for their antidiarrhoeal activity. Most studies evaluated their effect on intestinal motility and antimicrobial activity and, therefore, did not take into account the pathogenesis of infectious diarrhoea. Features of infectious diarrhoea like abdominal pain, cramps, inflammation, and passage of blood/mucus in the stools are the combined effect of one or more virulence factors of the infecting organism. The effect of medicinal plants on the microbial virulent features can serve as marker(s) for testing their efficacy. In this study, we evaluated the effect of a decoction of dried leaves of Dalbergia sissoo on aspects of pathogenicity, that is, colonisation to intestinal epithelial cells and production/action of enterotoxins. This was done to define its possible mechanism(s) of action in infectious diarrhoea. Materials and Methods : Antibacterial, antiprotozoal, and antiviral activities of the plant decoction were checked by agar dilution method, tube dilution method, and neutral red uptake assay, respectively. Cholera toxin (CT) and Escherichia coli labile toxin (LT) were assayed by ganglioside monosialic acid receptor ELISA. Suckling mouse assay was used to assess E. coli stable toxin (ST). As a measure of colonisation, the effect against adherence of E. coli and invasion of E. coli and Shigella flexneri to HEp-2 cells were studied. Results: The decoction had no antibacterial, antiprotozoal, and antiviral activity. It reduced the production and the binding of CT and bacterial adherence and invasion. Conclusion : This study showed that D . sissoo is antidiarrhoeal as it affects bacterial virulence. However, it has no antimicrobial activity.
  • ... Adherence also aids in subsequent proliferation and colonization of the gut and may be followed by toxin production or invasion [13]. The importance of using colonization and production and action of enterotoxins as specific parameters reflecting the pathogenesis has been earlier used by us as an approach towards understanding the varied mechanism(s) of action of antidiarrhoeal medicinal plants against infectious diarrhoea [14-17]. The studies highlighted the necessity of including multiple parameters for assessing effectiveness of medicinal plants against infectious forms of diarrhoea, especially in the absence of antimicrobial activity. ...
    Article
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    Aegle marmelos (L.) Correa has been widely used in indigenous systems of Indian medicine due to its various medicinal properties. However, despite its traditional usage as an anti-diarrhoeal there is limited information regarding its mode of action in infectious forms of diarrhoea. Hence, we evaluated the hot aqueous extract (decoction) of dried unripe fruit pulp of A. marmelos for its antimicrobial activity and effect on various aspects of pathogenicity of infectious diarrhoea. The decoction was assessed for its antibacterial, antigiardial and antirotaviral activities. The effect of the decoction on adherence of enteropathogenic Escherichia coli and invasion of enteroinvasive E. coli and Shigella flexneri to HEp-2 cells were assessed as a measure of its effect on colonization. The effect of the decoction on production of E. coli heat labile toxin (LT) and cholera toxin (CT) and their binding to ganglioside monosialic acid receptor (GM1) were assessed by GM1-enzyme linked immuno sorbent assay whereas its effect on production and action of E. coli heat stable toxin (ST) was assessed by suckling mouse assay. The decoction showed cidal activity against Giardia and rotavirus whereas viability of none of the six bacterial strains tested was affected. It significantly reduced bacterial adherence to and invasion of HEp-2 cells. The extract also affected production of CT and binding of both LT and CT to GM1. However, it had no effect on ST. The decoction of the unripe fruit pulp of A. marmelos, despite having limited antimicrobial activity, affected the bacterial colonization to gut epithelium and production and action of certain enterotoxins. These observations suggest the varied possible modes of action of A. marmelos in infectious forms of diarrhoea thereby validating its mention in the ancient Indian texts and continued use by local communities for the treatment of diarrhoeal diseases.
  • Article
    Bacterial adhesion is the first step in the sequence of events leading to infection. Previous data are available on the effect of Holarrhena antidysenterica on antidiarrhoeal and antibacterial action, but there is little information on the mechanism of action of the various aspects of EPEC-induced diarrhoea, namely adherence and translocation of the effector molecule to intestinal epithelial cells. The aim of the present study was to investigate the effects of alkaloids of Holarrhena antidysenterica (AHA) on interference in the mechanism of enteropathogenic Escherichia coli (EPEC) adhesion on host epithelial cells (INT 407 and HEp2). To determine the impact of AHA on epithelial cells, cytotoxicity (LDH), adherence, apoptotic and ultrastructural studies were performed. To analyse the effect of AHA on EPEC secreted proteins, especially EspD, INT 407 monolayers were infected with EPEC and AHA-treated EPEC, followed by immunoblotting, probed with anti EspD antisera. The maximum percentage of LDH leakage was reduced in AHA-treated EPEC (400 microg/mL) in both cell lines. Reduced bacterial adherence was observed under light microscopy and altered apoptotic changes were visualized using propidium iodide staining in conjunction with fluorescence microscopy, in both cell lines infected with AHA-treated EPEC and these results were confirmed with transmission electron microscope images. The suppression of type III secretory proteins (TTSPs), EspD ( approximately 40 kDa), was detected in INT 407 cells infected with AHA-treated EPEC. In conclusion, AHA reduces initial bacterial adhesion to intact epithelial cells and it may exert an antiadherence effect against the pathogenesis of EPEC in host epithelial cells. Thus, the investigations provide a rational basis for the treatment of EPEC-mediated diarrhoea with AHA.
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    While data are available on the effect of medicinal plants on intestinal motility and their antibacterial action, there is a paucity of information on their mode of action on various aspects of diarrheal pathogenicity, namely colonization to intestinal epithelial cells and production/action of enterotoxins. Crude decoction of dried leaves of Pongamia pinnata was evaluated for its antimicrobial (antibacterial, antigiardial and antirotaviral) effect; and its effect on production and action of enterotoxins (cholera toxin, CT; Escherichia coli labile toxin, LT; and E. coli stable toxin, ST); and adherence of enteropathogenic E. coli and invasion of enteroinvasive E. coli and Shigella flexneri to epithelial cells. The decoction had no antibacterial, antigiardial and antirotaviral activity, but reduced production of CT and bacterial invasion to epithelial cells. The observed results indicated that the crude decoction of P. pinnata has selective antidiarrheal action with efficacy against cholera and enteroinvasive bacterial strains causing bloody diarrheal episodes.
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    Previously described GM1 ganglioside enzyme-linked immunosorbent assays (GM1-ELISA) for the detection of Escherichia coli heat-labile enterotoxin (LT) showed sensitivity equal to the Y-1 adrenal cell assay when anti-LT (a reagent not commercially available) was used. However, when antitoxin to immunologically related (commercially available) cholera toxin was substituted, a marked loss in sensitivity occurred. We modified the GM1-ELISA that employed anti-cholera toxin to make it comparable in sensitivity to the Y-1 adrenal cell assay. When five media commonly used for LT production were compared, Mundell's Casamino Acids medium was shown to be significantly superior. Lincomycin (45 micrograms/ml) added to E. coli cultures significantly increased net optical densities in the GM1-ELISA, a direct measure of the amount of LT. Treatment of broth cultures or bacterial cell pellets with polymyxin B or extension of culture time to 48 h also significantly increased net optical density by allowing enhanced release of periplasmic LT. A major innovation involved the direct culture of E. coli strains in GM1-coated wells of microtiter plates followed by ELISA. This direct culture method GM1-ELISA (DCM-GM1-ELISA) saved not only assay time, but also materials and reagents. The net optical densities that result from this assay allow the test to be read visually without a spectrophotometer. Three independent observers read plates with E. coli tested by DCM-GM1-ELISA. Thirty-four of 35 adrenal cell-positive strains (97% sensitivity) and 30 of 30 LT-negative control E. coli strains (100% specificity) were identified by all three observers reading coded plates. The DCM-GM1-ELISA provides a simple, practical and efficient assay for LT for less sophisticated laboratories.
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