子痫前期血清通过PLCγ1促进脐血管平滑肌细胞PKC-α活性及胶原的表达

Preeclampsia sera induces HUASMC PKC-α activation and precollagen Ⅰexpression via PLC-γ1 pathway

目的:探讨子痫前期(PE)血清激活血管平滑肌细胞蛋白激酶C-α(PKC-α)及前胶原Ⅰ表达是否由磷脂酶C-γ1(PLC-γ1)信息分子介导。方法:原代培养脐静脉内皮细胞(HUVEC)与脐动脉平滑肌细胞(HUASMC),体外建立HUVEC与HUASMC共培养体系,用PE血清处理共培养HUASMC细胞。MTT法检测HUASMC细胞活力;Annexin VFITC-流式细胞仪检测HUASMC凋亡;Western blot法检测PLC-γ1(磷酸化)、PKC-α(包膜/胞浆比值)活性以及前胶原Ⅰ表达。siRNA沉默PLC-γ1表达后,观察PE血清对共培养HUVEC细胞PKC-α活性及前胶原Ⅰ表达的影响。结果:PE血清可促进共培养HUASMC细胞PLC-γ1磷酸化、PKC-α膜转位及前胶原Ⅰ的表达(P<0.05);siRNA沉默HUASMC细胞PLC-γ1蛋白表达可逆转PE血清引起的PKC-α膜转位及前胶原Ⅰ的表达(P<0.05)。结论:PE血清通过PLC-γ1-PKC-α信息途径促进HUASMC前胶原Ⅰ的表达,可能在胎盘血管病理过程中起着重要的作用。

Objective: To explore whether preeclampsia sera induces HUASMC PKC-α activation and precollagen Ⅰ expression via PLC-γ1 pathway. Methods: HUVEC and HUASMC were co-cultured in transwells with preeclamptic sera. PLC-γ1 siRNA was transferred into cocultured HUASMC before incubated with preeclamptic sera. The co-cultured HUASMC incubated with preeclampsia sera,normal pregnancy sera was used as control. The cell viability was detected by MTT. The apoptosis of HUASMC were analyzed by flow cytometry. The PLC-γ1 protein activity,the expression of particulate fraction and cytoplasmic protein kinase C-α( PKC-α) were detected by Western blot after HUASMC incubated with preeclampsia sera for 24h. Results: Preeclamptic sera could stimulate the cell viability,the activation of PKC-α,the activation of PLC-γ1,and the expressions of precollagen Ⅰ of co-cultured HUASMC as compared with normal pregnancy sera( P < 0. 05). The PLC-γ1 silence of co-cultured HUASMC decreased the cell viability,the activation of PKC-α,and the precollagen Ⅰ expressions of co-cultured HUASMC stimulated by preeclamptic sera( P < 0. 05). Conclusion: Preeclampsia sera induces HUASMC PKC-α activation and precollagen Ⅰexpression via PLC-γ1 pathway,which could be more important in the pathogenesis of placental blood vessel in preeclampsia.