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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 To investigate the effect of serum of preeclampsia on the changes of calcium ion concentration in umbilical artery smooth muscle cells and the regulation of phospholipase C-γ1 (PLC-γ1) -inositol triphosphate receptor (IP3R) pathway. Methods The co-culture system of endothelial cells and smooth muscle cells was established in vitro. The co-cultured cells were treated with serum of preeclampsia, the activity of smooth muscle cells was detected by MTT assay, and the activity of PLC-γ1 (phosphorylation), IP3R, Fluo The effect of preeclampsia serum on IP3R and intracellular Ca2 + concentration in smooth muscle cells was observed with PLC-γ1siRNA silencing smooth muscle cells (PLC-γ1). Results Serum of preeclampsia promoted the phosphorylation of PLC-γ1, IP3R and the concentration of calcium ion in co-cultured smooth muscle cells. The silencing of PLC-γ1 in smooth muscle cells reversed the IP3R activity and the increase of intracellular calcium in co-cultured smooth muscle cells induced by preeclampsia . Conclusions Serum of preeclampsia promotes the increase of intracellular Ca2 + in umbilical artery smooth muscle cells via PLC-γ1-IP3R signaling pathway. The increase of intracellular Ca2 + in smooth muscle cells induced by activation of PLC-γ1-IP3R may play an important role in the pathological process of preeclampsia vascular smooth muscle cells .