The Apoptosis Pathway Of Jiangtangsanhuang Tablet On The Rat Insulin Beta-Cell

Objective:We discussed the death receptors on Fas/FasL pathways by the effect of different doses of Jiangtangsanhuang tablet on the rats islet beta cells induced by interleukin-1β to elaborate the mechanism of alleviating the apoptosis by Jiangtangsanhuang tablet.MethodsIt was obtained normal islet beta cells from SD rats by the methods of bile reflux perfusion and removaling perfused pancreata. The methods of MTT assay, RNA isolation and real-time PCR of fasmRNA, baxmRNA and bcl-2mRNA, Western Blot of proteins with fasl, cyto-c and caspase-3were also used to complete our experiment.Results1. By measuring the insulin release quantity, It was obtained normal function islet cells from rats with11.1mmol/L glucose concentration of cell culture.2. Islet cells pretreated wih IL-1β for24h with the dose concentrations from50μg/ml to200iμ/ml could protect the life activity of islet cells selected by MTT assay method(P<0.05).3. Islet cells pretreated wih IL-1β for24hours and RT-PCR found that the expression of FasmRNA and BaxmRNA in the stimulated group by IL-1β increased by comparing with the blank group (P<0.05) while the expression of Bcl-2mRNA reduced (P<0.05). After24hours of drug intervention, comparing with the stimulated group by IL-1β, the expression of FasmRNA in middle concentration of Jiangtangsanhuang tablet group, and high concentration of Jiangtangsanhuang tablet group and pioglitazone group decreased (P<0.05) while low concentration of Jiangtangsanhuang never observe the decline(P>0.05). Multiple group comparisons by LSD found high concentration of Jiangtangsanhuang tablet group had better effect than pioglitazone group (P <0.05), pioglitazone group had better effect than middle dose of Jiangtangsanhuang tablet group (P<0.05). After24hours of drug intervention, comparing with the stimulated group by IL-1β, the expression of BaxmRNA in middle concentration of Jiangtangsanhuang tablet group, and high concentration of Jiangtangsanhuang tablet group and pioglitazone group decreased (P<0.05) while low concentration of Jiangtangsanhuang never observe the decline(P>0.05). Multiple group comparisons by LSD found the result with middle concentration of Jiangtangsanhuang tablet group, and high concentration of Jiangtangsanhuang tablet group and pioglitazone group had no significant differences(P>0.05). After24hours of drug intervention, comparing with the stimulated group by IL-1β, the expression of Bcl-2mRNA in middle concentration of Jiangtangsanhuang tablet group, and high concentration of Jiangtangsanhuang tablet group and pioglitazone group increased (P<0.05) while low concentration of Jiangtangsanhuang never observe the ascend(P<0.05). Multiple group comparisons by LSD found high concentration of Jiangtangsanhuang tablet group had more effect than pioglitazone group(P<0.05), pioglitazone group had less effect than middle dose of Jiangtang sanhuang tablet group (P<0.05).4. Put in after24h,Use Western Blot found islet cells induced by IL-1β expressed more proteins of FasL, Cyto-c, caspase-3than the blanked group. After the drug intervention, it showed that high concentration of Jiangtangsanhuang tablet group, middle concentration of Jiangtangsanhuang tablet group and pioglitazone group more reduced the protein expression of Fasl, Cyto-c and caspase-3than the blanked group while, low concentration of Jiangtangsanhuang tablet group was not obviously decreased by comparing with blank group.ConelusionIn our present study, we have evidences that Jiangtangsanhuang tablet can inhibit expression of Fas/Fasl and Cyto-C release, decline Bax and raise Bcl-2to protect rat islet cells fromβ-cell damage induced IL-1β which maybe the preventive mechanism of Jiangtangsanhuang Tablet to suppress the apoptosis of β-cells.