Abstract
Objective:To investigate the effect of Tangeretin(TAN)on high glucose(HG)-induced apoptosis in trophoblast cells by regulating the Ras homologous gene family member A(RhoA)/Rho-associated coiled-coil forming protein kinase(ROCK)signaling pathway.Methods:Human chorionic trophoblast cells(HTR8-SVneo)were cultured in vitro and randomly divided into control group(conventional culture),HG group(25 mmol/L glucose),L-TAN,M-TAN,H-TAN groups(25,50,and 100 μg/mL TAN,respectively),and PMA group(100 μg/mL TAN+100 ng/mL RhoA/ROCK signaling pathway activator PMA).MTT method and colony formation assay were applied to detect cell proliferation.ELISA was applied to detect the levels of interleukin(IL)1β(IL-1β),tumor necrosis factor-α(TNF-α),IL-6 and IL-8.Flow cytometry was applied to detect apoptosis.Western blotting was used to detect the expression of proliferating cell nuclear antigen(PCNA),apoptosis-associated proteins(Bcl-2,Bax,cleaved caspase-3),and proteins related to the RhoA/ROCK signaling pathway(RhoA,ROCK).Results:Compared with the control group,the A490 of cells,clone number,the PCNA,Bcl-2 protein expression in the HG group were decreased,while the IL-1β,TNF-α,IL-6,IL-8,apoptosis rate,cleaved caspase-3,Bax,RhoA,ROCK protein expression were elevated.Compared with the HG group,the A490 of cells,clone number,the PCNA,Bcl-2 protein expression in the L-TAN,M-TAN,and H-TAN groups were increased,while the IL-1β,TNF-α,IL-6,IL-8,apoptosis rate,cleaved caspase-3,Bax,RhoA,ROCK protein expression were decreased.Compared with H-TAN group,the A490 of cells,clone number,the PCNA,Bcl-2 protein expression in PMA group were decreased,while the IL-1β,TNF-α,IL-6,IL-8,apoptosis rate,cleaved caspase-3,Bax,RhoA,ROCK protein expression were elevated.Conclusion:Tangeretin may attenuate high glucose-induced inflammatory responses in trophoblast cells by inhibiting the RhoA/ROCK signaling pathway,thereby promoting cell proliferation and inhibiting cell apoptosis.关键词
橘皮素/Ras同源基因家族成员A/Rho相关卷曲螺旋形成蛋白激酶信号通路/滋养层细胞/细胞凋亡/妊娠期糖尿病Key words
Tangeretin/Ras homolog gene family member A/Rho-associated coiled-coil forming protein kinase signaling pathway/trophoblast cell/apoptosis/gestational diabetes mellitus分类
基础医学
