The Therapeutic Effects And Mechanism Of Chinese Medicine Plantaginis Semen And Its Active Components On Salmonella Typhimurium

Purpose:This study investigates the therapeutic effects of plantaginis semen and its active components against salmonella typhimurium in mice.Furthermore,the therapeutic effects and the mechanisms of immune intervention are investigated by molecular biological method and cytological method.Method:1.Select 80 Kunming male mice into 2 groups and feed with different amount of salmonella typhimurium.Mortality,growth condition and pathological changes of mice are observed for the protective effect of plantaginis semen and its active components on the infection of salmonella typhimurium in mice.2.The contents of the main active components?geniposidic acid,verbascoside and aucubin?of extract of plantaginis semen are determined by liquid chromatography.The minimal inhibitory concentration of plantaginis semen and its active components against salmonella typhimurium is determined by single concentration broth microdilution recommended by NCCLS,the OD₆₀₀00 value of salmonella typhimurium bacteria liquid,the growth conditions of salmonella typhimurium are observed and the growth curves are analyzed3.Calculating the invasion rate of salmonella typhimurium invasion of Hela cells,the inhibition of plantaginis semen and its active components on salmonella typhimurium is determined.The cytotoxic effects of extract of plantaginis semen are determined by the absorption values of 490nm using the enzyme mark instrument,and the results of calculating the release amount of LDH from the supernatant of the samples.4.Verify the inhibition effect of the main active components of plantaginis semen for the main effect protein SipA and SipC of salmonella typhimurium's invasion by western blot method.The therapeutic effects and the mechanisms of immune intervention of plantaginis semen and aucubin are investigated.Result:1.The extract of plantaginis semen can protect the mice in morality and weight loss during the infection of salmonella typhimurium.For comparison,the structure of goblet cells of salmonella typhimurium treatment mice group's caecum damaged or disappeared,neutrophil infiltrations are observed.Subcutaneous edemas of mice's caecum are not obvious in medical solution treatment group and inva knockout group,goblet cells'structure keep intact.2.The content of geniposide acid is 0.76%,the content of verbascoside is 0.51%and the content of acubin is 0.04%.And the components have no inhibition effect of the growth of salmonella typhimurium in normal effective drug concentration.3.When the effective drug concentration is 16mg/mL and 32mg/mL,the cell viability of Hela cells has no effects,and can effectively inhibit the salmonella typhimurium invasion of Hela cells.The cytotoxic effect is observed when the effective drug concentration is128mg/mL.Results show that the extract of plantaginis semen can efficiently inhibit the salmonella typhimurium invasion on epithelial cells and the intervention in cell infection.4.The extracts plantaginis semen have no inhibition effects on the gene expression of SipA.But plantaginis semen and the main active components aucubin can efficiently inhibit the gene expression of the main effect protein SipC.Results show that plantaginis semen can prevent the salmonella typhimurium invasion by deducing SipC's functions,and the main effective component is aucubin.Conclusion:Plantaginis semen and its main active component aucubin can effectively inhibite the expression of SipC,deducing the function of SPI-1 and weakened salmonella typhimurium invasion ability.Plantaginis semen and its active components can also protect the mice from infection.The therapeutic effects of plantaginis semen and its active components are verified and the mechanisms of immune intervention are further confirmed.Thus,based on the traditional chinese medicine theory of clearing heat,infiltrating moisture and antidiarrheal function of plantaginis semen,this study provides a theory and experimental base for the traditional chinese clinical practice.