Protective Effect Of Water Extract Of Clematis On INS-1 Cells’ Apoptosis Induced By PCBs

Objective:To observe the protective effect of different concentration of water extract of Clematis（Clematis）in rat insulinoma INS-1 cells’ apoptosis induced by PCBs（PCB118）.Methods:The INS-1 cell lines were divided into control group（control）,PCB118 group（25μM PCB118）,low concentration Clematis group(25 μM PCB118 +4mg·L^-1 Clematis)and high concentration Clematis group(25μM PCB118 +40mg·L^-1 Clematis).INS-1 cell was treated with different concentration of Clematis and PCB118 as mentioned above,the apoptosis situation and morphological change of ISNS-1 were observed by inverted microscope,the activity of INS-1 cells was assayed by CCK-8 method,cell apoptosis rate was further detected by flow cytometry,the Western blot analysis was used to measure the expression of Cleaved Caspase-3、Caspase-3.Results:After PCB118 9h,24h,48h stimulation,INS-lcells showed obvious apoptotic morphology.With co-stimulation with high or low concentration of Clematis the apoptosis rate decreased;CCK-8 assay result indicated that cell proliferation and activity slowed after PCB118 treated（P<0.01）,while the two Clematis groups reserved cell activity inhibited by PCB118.The difference was statistically significant（P<0.05）when high concentration Clematis group compared with apoptosis group.Flow cytometry showed that PCB118 could significantly induce INS-1 cells’ early apoptosis（P<0.05）.With the intervention of Clematis,the early apoptosis was improved,whereas high concentration group showed stronger protective effect（P<0.05）.After PCB118 treatment,the expression of apoptosis-related protein Cleaved Caspase-3 was upregulated.Clematis could reverse the overexpression of Cleaved Caspase-3 induced by PCB118.Conclusion:Water extract of Clematis can inhibit apoptosis of INS-1 cells induced by PCB118.Clematis enhance the activity of INS-1 cells in PCB118 toxic environment.Our result will provide experimental evidence for TCM to protect islet β cells.