子痫前期血清通过PLCγ1-IP_3R信息途径促进脐动脉平滑肌细胞内钙离子聚集

Preeclampsia sera induces intracellular Ca^(2+) Concentration in HUASMC via PLC-γ1-IP_3R pathway

目的探讨子痫前期血清对脐动脉血管平滑肌细胞内钙离子浓度变化的影响以及磷脂酶C-γ1(PLC-γ1)-三磷酸肌醇受体(IP3R)信息途径对其的调控作用。方法体外建立内皮细胞与平滑肌细胞共培养体系,子痫前期血清处理共培养细胞,MTT检测平滑肌细胞活力,Western blot检测PLC-γ1(磷酸化)、三磷酸肌醇受体(IP3R)活性,Fluo-3/AM检测平滑肌细胞内钙离子浓度,PLC-γ1siRNA沉默平滑肌细胞PLC-γ1表达后,观察子痫前期血清对共培养平滑肌细胞IP3R及细胞内钙离子浓度变化的影响。结果子痫前期血清可促进共培养平滑肌细胞PLC-γ1、IP3R磷酸化及钙离子浓度,平滑肌细胞PLC-γ1沉默可逆转子痫前期血清引起的共培养平滑肌细胞内IP3R活性及细胞内钙离子增加。结论子痫前期血清通过PLC-γ1-IP3R信息通路促进脐动脉平滑肌细胞内钙离子增加,PLC-γ1-IP3R激活引起平滑肌细胞内钙离子增加可能在子痫前期血管平滑肌细胞病理过程中起重要作用。

Objective： We tested whether Preeclampsia Sera Induces intracellular Ca^2＋ concentration in HUASMC via PLC- γ1-IP3R pathway. Methods： HUVEC and HUASMC were co-cultured in transwells with preeclamptic sera. PLC-γ1 siRNA was transferred into co-cultured HUASMC before incubated with preeclamptic sera. The co-cultured HUASMC incubated with normal pregnancy sera was as control. The cell viability was detected by MTT. The apoptosis of HUASMC were analyzed by flow cytomerty. The PLC- γ1 and IP3 R protein activity were detected by Western blotting and the intracellular Ca^2＋ concentration in VSMC was monitored using the fluorescent Ca^2＋ indicator, Fluo-3/AM after HUASMC was incubated for 24h. Results- Preeclarnptic sera could stimulate the cell viability, the activation of PLC- γ 1 and IP3R, and the intracellular Ca^2＋concentration in co-cultured HUASMC as compared with normal pregnancy sera （P〈0.05） .The PLC-γ 1 silence of co-cultured HUASMC decreased the cell viability, the activation of IP3R, and the intracelhilar Ca^2＋ concentration in co-cultured HUASMC stimulated by preeclamptic sera （P〈0.05） . Conclusion： Preeclampsia Sera Induces intracellular Ca^2＋ concentration in huasmc in co-cultured HUASMC via PLC- γ1-IP3R pathway. PLC- γ1-IP3R signal transduction pathway may play a key role in SMC construction, which is more important in the pathogenesis of placental blood vessel in preeclampsia.