ABSTRACT
The natural products derived from medicinal plants are proven to be the abundant source of biologically active compounds, which have been the basis of development of new pharmaceuticals. The present study was carried out to understand the antibacterial activity of methanolic extract of Punica granatum L. seed (MPG) against the clinically isolated urinary tract infections (UTI) causing bacteria i.e. Escherichia coli and Klebsiella pneumoniae using different methods i.e. disc diffusion, agar well diffusion, modified agar well diffusion. Colony forming units per ml (CFU/ml) were also determined for wild and MPG treated bacteria by spread plate and absorbance method. Phytochemical analysis of MPG and biochemical characterization of K. pneumoniae and E. coli was carried out using a standard protocol. MPG was a dark red colour sticky mass with a yield of 26.87% (w/w). Phytochemical analysis revealed the presence of steroids, tannins, coumarins, glycosides, flavonoids and phenolic compounds. The present investigation showed that MPG was effective against K. pneumoniae and E. coli. K. pneumoniae was a multi-drug resistant strain, found to be more sensitive to MPG in comparison to E. coli. The CFU/ml was found to be reduced in a dose-dependent manner in MPG treated urobacteria, which suggests that P. granatum seeds can be used as a remedy for UTI and also can be taken regularly to prevent the recurrence of UTI.
ABSTRACT
BACKGROUND & OBJECTIVE: Previous studies on natural products had mainly dealt with their antimicrobial activity and studies on the interference of these bioactive compounds with host-bacterial interaction is limited. The present study was undertaken to investigate the effect of the sterols and fatty acids present in the chloroform fraction of crude methanol extract of Hemidesmus indicus root (CHI) on Salmonella enterica serovar Typhimurium (S. Typhimurium) mediated apoptosis in a murine macrophage cell line (P388D1). METHODS: Bacterial sensitivity test was carried out with different concentrations of CHI and the optimum dose was fixed as 100 mug/ml for CHI, which was safe on host cells as the CD(50) (50% of cell death) dose of CHI was determined to be 500 mug/ml in the P388D1 cell line. RESULTS: The CHI-treated bacteria had negligible cytotoxicity and were less potent to invade and proliferate intracellularly. Murine macrophages infected with wild bacteria, stained with Hoechst 33258, had swollen and damaged morphology with characteristic apoptotic bodies whereas macrophages infected with treated bacteria had comparative normal architecture. Immunofluorescence and transmission electron micrographs both confirmed that CHI-treated bacteria were defective and smaller than the wild bacteria. Ultrastructures of P388D1 cells infected with wild bacteria showed many ingested bacteria and characteristic Salmonella-containing vacuoles (SCV). Some cells had condensed or fragmented nuclei with swollen mitochondria, whereas most of the cells infected with treated bacteria were normal in morphology and a few had internalized bacteria, but the typical bacteria laden SCV was not observed in cells infected with CHI-treated S. Typhimurium. INTERPRETATION & CONCLUSION: Our results showed that the choloroform fraction of H. indicus root blocked the cytotoxic activity of S. Typhimurium in a macrophage cell line. More studies need to be done to elaborate and confirm our findings.
Subject(s)
Animals , Apoptosis/drug effects , Cell Line , Cell Proliferation/drug effects , Hemidesmus , Macrophages/microbiology , Mice , Plant Extracts/pharmacology , Plant Roots , Salmonella typhimurium/drug effects , VirulenceABSTRACT
Background & objectives: Previous studies on natural products had mainly dealt with their antimicrobial activity and studies on the interference of these bioactive compounds with host-bacterial interaction is limited. The present study was undertaken to investigate the effect of the sterols and fatty acids present in the chloroform fraction of crude methanol extract of Hemidesmus indicus root (CHI) on Salmonella enterica serovar Typhimurium (S. Typhimurium) mediated apoptosis in a murine macrophage cell line (P388D1). Methods: Bacterial sensitivity test was carried out with different concentrations of CHI and the optimum dose was fixed as 100 μg/ml for CHI, which was safe on host cells as the CD50 (50% of cell death) dose of CHI was determined to be 500 μg/ml in the P388D1 cell line. Results: The CHI-treated bacteria had negligible cytotoxicity and were less potent to invade and proliferate intracellularly. Murine macrophages infected with wild bacteria, stained with Hoechst 33258, had swollen and damaged morphology with characteristic apoptotic bodies whereas macrophages infected with treated bacteria had comparative normal architecture. Immunofluorescence and transmission electron micrographs both confirmed that CHI-treated bacteria were defective and smaller than the wild bacteria. Ultrastructures of P388D1 cells infected with wild bacteria showed many ingested bacteria and characteristic Salmonella-containing vacuoles (SCV). Some cells had condensed or fragmented nuclei with swollen mitochondria, whereas most of the cells infected with treated bacteria were normal in morphology and a few had internalized bacteria, but the typical bacteria laden SCV was not observed in cells infected with CHI-treated S. Typhimurium. Interpretation & conclusions: Our results showed that the choloroform fraction of H. indicus root blocked the cytotoxic activity of S. Typhimurium in a macrophage cell line. More studies need to be done to elaborate and confirm our findings.
ABSTRACT
Methanolic extract of H. indicus root (MHI) was screened for its antimicrobial activity against S. typhimurium, E. coli and S. flexneri, in vitro and in experimentally induced diarrhoea in albino rats, in vivo. MHI had an anti enterobacteriae effect as evident from agar well diffusion method and decrease in CFU/ml in MHI treated LB broth culture. MHI inhibited the castor oil induced diarrhoea in rats as judged by a decrease in the amount of wet faeces in MHI-pretreated rats at a dose of 500-1500 mg/kg. The results indicated that MHI was more active than standard antidiarrhoeal drug, lomotil. Phytochemical tests revealed the main constituents as tannins, steroids, triterpenoids and carbohydrates. Present findings suggested that MHI might elicit an antidiarrhoeal effect by inhibition of intestinal motility and by its bacteriocidal activity.