摘要
目的:观察不同浓度威灵仙水提物(Clematis)对多氯联苯(PCB118)诱导的胰岛β细胞凋亡的保护作用。方法:将大鼠胰岛素瘤(INS-1)细胞株分为对照组(control)、PCB118(25μmol/L)组、Clematis低剂量组(4mg/L)以及Clematis高剂量组(40mg/L),分别于药物干预9、24、48h后于倒置显微镜下观察INS-1细胞形态,通过CCK-8法检测各组INS-1细胞活性,并采用流式细胞术检测细胞的凋亡率,Western Blot检测Cleaved Caspase-3、Caspase-3蛋白的表达。结果:倒置显微镜下观察结果显示:PCB118刺激9、24、48h后INS-1细胞凋亡,Clematis高浓度及低浓度组PCB118诱导的INS-1细胞凋亡减少;CCK-8结果显示PCB118处理后细胞活性显著下降(P<0.01),而Clematis高、低浓度组均改善PCB118的抑制作用,其中高浓度组与PCB118组比较,差异有统计学意义(P<0.05);流式细胞术检测结果显示PCB118诱导INS-1细胞早期凋亡明显(P<0.05),而Clematis干预后细胞早期凋亡改善,高浓度组更为显著(P<0.05);同时,PCB118处理后凋亡相关蛋白Cleaved Caspase-3表达上调,Clematis可逆转Cleaved Caspase-3过表达(P<0.05)。结论:威灵仙水提物可以抑制PCB118诱导的INS-1细胞凋亡,增强PCB118毒性环境下的INS-1细胞活性,为中药保护胰岛β细胞提供了实验依据。
Objective: To observe the protective effect of different concentration of Clematis water extract in rat insulinoma INS-1 cells' apoptosis induced by PCB118. Methods: The INS-1 cell lines were divided into control group(control), PCB118(25μmol/L) group, PCB118+Clematis(4mg/L) group and PCB118+Clematis(40mg/L) group. INS-1 cell was treated with different concentration of Clematis and PCB118 as mentioned above, the morphological change and apoptosis situation of INS-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, and the expression of Cleaved Caspase-3, Caspase-3 was measured by Western blot analysis. Results: After PCB118 9, 24, 48 h stimulation, INS-1 cells 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(P<0.05). 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.
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2017年第10期4616-4619,共4页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
江苏省自然科学基金项目(No.BK20151591)
国家自然科学基金项目(No.81270861)~~
