芍药苷对皮质酮致原代皮层细胞神经损伤的保护作用及机制研究

Effects and mechanism of paeoniflorin on cell model of never injury induced by corticosterone

目的:探讨促丝裂原活化蛋白激酶(MEK)/细胞外信号调节激酶(ERK)信号通路在芍药苷拮抗皮质酮诱导的神经损伤作用及其机制,为抑郁症的预防与治疗提供新思路。方法:采用高浓度皮质酮制备原代皮层神经细胞抑郁症及神经损伤模型,细胞随机分为正常组、皮质酮组、芍药苷(1、10、50μmol/L)组、芍药苷(1、10、50μmol/L)+皮质酮组、U0126/PD98059组+正常组、U0126/PD98059组+皮质酮组、U0126/PD98059+芍药苷+皮质酮组,共14组。进行细胞纯度鉴定,观察细胞形态,检测细胞存活率,检测细胞ERK1、ERK2、CREB m RNA的表达,检测细胞p-ERK/ERK、p-CREB/CREB蛋白的表达。结果:培养的细胞胞体饱满,状态良好,纯度较高。与正常组比较,皮质酮组可降低细胞存活率(P<0.01),芍药苷各剂量组可不同程度使细胞存活率增加(P<0.01,P<0.05),U0126或PD98059能逆转芍药苷提高细胞存活率的作用。与正常组比较,皮质酮组细胞ERK1、ERK2、CREB m RNA含量显著降低(P<0.01,P<0.05);与皮质酮组比较,芍药苷50μmol/L+皮质酮组细胞ERK1 m RNA及芍药苷10μmol/L+皮质酮组细胞中ERK2、CREB m RNA含量显著升高(P<0.05)。与正常组比较,皮质酮组细胞p-ERK/ERK、p-CREB/CREB蛋白表达均下降(P<0.01);与皮质酮组比较,芍药苷各剂量组细胞p-ERK/ERK蛋白表达水平呈显著性升高(P<0.01);与芍药苷10μmol/L+皮质酮组比较,加入U0126/PD98059预处理后,细胞p-ERK/ERK、p-CREB/CREB蛋白表达水平下降。结论:芍药苷能拮抗皮质酮诱导的原代皮层神经细胞的神经损伤,其作用机制由MEK/ERK信号通路介导。

Objective： To investigate the antagonism effect and mechanism of paeoniflorin of the MEK/ERK signaling pathway in the nerve injury induced by corticosterone and provide new ideas for the prevention and treatment of depression. Methods： The nerve injury and depression model was established in the primary cortex nerve cells using high levels of corticosterone, the cells were randomly divided into control group, corticosterone group, paeoniflorin group （1, 10, 50μmol/L）, paeoniflorin （1, 10, 50μmol/L）＋eorticosterone group, U0126/PD98059＋control group, U0126/PD98059＋corticosterone group and U0126/PD98059＋paeoniflorin＋corticosterone group, 14 groups in total. The cell purity was identified, the cell morphology was observed, and the cell survival rate, mRNA expressions of ERK1, ERK2 and CREB, protein expressions of p-ERK/ERK and p-CREB/CREB were detected. Results： The Cell was in the good shape, with high purity. Corticosterone could lead to the decrease of cell survival rate （P〈0.01）, and paeoniflorin could increase the cell survival rate in each dose group （P〈0.01, P〈0.05）, but.UO126 or PD98059 could reverse the effect of improvement of cell survival rate by paeoniflorin （P〈0.01, P〈0.05）. Compared with control group, the mRNA levels of ERK1, ERK2 and CREB in the corticosterone group were significantly decreased （P〈0,01, P〈0.05）. Compared with corticosterone group, the mRNA levels of ERK1, ERK2 and CREB were significantly increased in the groups with each dose paeoniflorin. Compared with control group, the p-ERK/ERK, p-CREB/CREB protein expressions weredecreased in the corticosterone group （P〈0.01）. Compared with corticosterone group, the p-ERK/ERK and p-CREB/CREB protein expressions were significantly increased in the paeoniflorin groups （P〈0.01）. Compared with paeonifl group pretreatment with U0126 or PD98059, the p-ERK/ERK and p-CREB/CREB protein expressions were decreased. Conclusion： Paeoniflorin has the antagonism effect of nerve injury in the primary cortical neural cells induced by corticosterone, and its mechanism is mediated by the MEK/ERK signaling pathway.