苦参碱调节Notch信号通路对高糖诱导的滋养层细胞增殖和凋亡的影响

Effect of matrine on the proliferation and apoptosis of high-glucose-induced trophoblast cells by regulating the Notch signaling pathway

目的研究苦参碱调节Notch信号通路而对高糖诱导的滋养层细胞增殖和凋亡产生的影响。方法将人绒毛膜滋养层细胞HTR-8/SVneo培养至对数期,以高糖诱导损伤,然后用不同浓度的苦参碱处理细胞并用CCK-8法检测其活性。依据结果将HTR-8/SVneo分为ctrl组(5 mmol/L葡萄糖培养)、HG组(25 mmol/L葡萄糖培养)、低浓度苦参碱组(1.00 mmol/L)、高浓度苦参碱组(1.50 mmol/L)、高浓度苦参碱(1.50 mmol/L)+Jagged1组(500μg/L Notch信号通路激活剂)。CCK-8试剂盒检测细胞增殖能力;流式细胞术检测细胞凋亡;试剂盒检测细胞中MDA和SOD活性;western blot检测Notch1、PCNA、BAX、Bcl-2蛋白表达。结果与ctrl组比较,HG组HTR-8/SVneo的细胞存活率、SOD水平、PCNA、Bcl-2蛋白表达降低,而细胞凋亡率、MDA水平、Notch1、BAX蛋白表达升高(P<0.05);与HG组比较,低浓度苦参碱组、高浓度苦参碱组细胞存活率、SOD水平、PCNA、Bcl-2蛋白表达升高,而细胞凋亡率、MDA水平、Notch1、BAX蛋白表达降低(P<0.05);Jagged1减弱了高浓度苦参碱对HTR-8/SVneo细胞增殖的促进和凋亡的抑制作用。结论苦参碱可能会通过抑制Notch信号通路而改善高糖诱导的滋养层细胞的增殖,并抑制细胞凋亡。

Objective To investigate the effect of matrine on the proliferation and apoptosis of high-glucose-induced trophoblast cells by regulating the Notch signaling pathway.Methods Human chorionic trophoblast cells HTR-8/SVneo were cultured to the logarithmic phase.They were induced with high glucose,and then treated with different concentrations of matrine.Cell viability was detected by CCK-8 assay.HTR-8/SVneo cells were induced with 5mmol/L glucose,25mmol/L glucose,1.00mmol/L matrine,1.50mmol/L matrine and 1.50mmol/L matrine+500μg/L Notch signaling pathway activator Jagged1.Cell proliferation and apoptosis were detected by CCK-8 assay and flow cytometry,respectively.Malondialdehyde(MDA)and superoxide dismutase(SOD)levels were detected by commercial kits.Protein expressions of Notch1,proliferating cell nuclear antigen(PCNA),BAX,B cell lymphoma-2(Bcl-2)were detected by Western blot.Results Compared with those induced with 5mmol/L glucose,HTR-8/SVneo cells induced with 25mmol/L glucose presented significantly lower cell survival rate,SOD level and protein expressions of PCNA and Bcl-2,but significantly higher apoptotic rate,MDA level and protein expressions of Notch1 and Bax(P<0.05).Compared with those induced with 25mmol/L glucose,HTR-8/SVneo cells induced with 1.00mmol/L or 1.50mmol/L matrine presented significantly higher cell survival rate,SOD level and protein expressions of PCNA and Bcl-2,but significantly lower apoptotic rate,MDA level and protein expressions of Notch1 and Bax(P<0.05).The treatment of Jagged1 significantly weakened the role of high-dose matrine in promoting the proliferation and inhibiting apoptosis of HTR-8/SVneo cells.Conclusion Matrine may improve the proliferation of high glucose-induced trophoblast cells and inhibit cell apoptosis by inhibiting the Notch signaling pathway.