论文部分内容阅读
目的观察MK801及SB203580对N-甲基-D-天(门)冬氨酸(NMDA)诱导的皮层神经元损伤的影响,探讨p38丝裂原活化蛋白激酶(p38MAPK)信号通路在神经元损伤中的作用和机制。方法培养7 d的新生SD大鼠皮层神经元随机分为对照组、NMDA损伤组、MK801干预组、SB203580干预组、联合干预组(SB203580+MK801)。MTT比色实验评价细胞生存力,乳酸脱氢酶(LDH)释放率测定细胞损伤程度,吖啶橙/溴化乙锭(AO/EB)双重荧光染色观察细胞凋亡形态和数目,Western blotting检测大鼠皮层神经元各组p38MAPK、BCL-2和BAX表达情况。结果与对照组比较,NMDA损伤组神经细胞存活力降低、培养上清液中LDH含量增加、凋亡细胞增多、p38MAPK和BAX表达增加(P均<0.01),BCL-2表达降低(P<0.05);与NMDA损伤组比较,各个干预组细胞生存力提高、LDH释放率降低、凋亡减少、p38MAPK及BAX的表达减少、BCL-2表达增多(P均<0.01)。结论 MK801,SB203580对NMDA诱导的皮质神经元损伤具有保护作用,p38MAPK通路可能参与介导MK801的保护作用,其共同机制是通过抑制p38MAPK信号通路介导的凋亡相关蛋白实现的。
Objective To investigate the effects of MK801 and SB203580 on N-methyl-D-aspartate (NMDA) -induced cortical neuron injury and to explore the role of p38 mitogen-activated protein kinase (p38MAPK) signaling pathway in neuronal damage The role and mechanism. Methods Cortical neurons of neonatal SD rats were randomly divided into control group, NMDA injury group, MK801 intervention group, SB203580 intervention group and combined intervention group (SB203580 + MK801). Cell viability was evaluated by MTT colorimetric assay. The degree of cell injury was determined by lactate dehydrogenase (LDH) release rate. Apoptosis morphology and number were observed by acridine orange / ethidium bromide (EB / EB) double staining. Expression of p38MAPK, BCL-2 and BAX in rat cortical neurons. Results Compared with the control group, the viability of nerve cells in NMDA injured group was decreased, the content of LDH in cultured supernatant increased, the number of apoptotic cells increased, the expression of p38MAPK and BAX increased (P <0.01) and the expression of BCL-2 decreased (P <0.05 ). Compared with NMDA group, the viability of all intervention groups increased, LDH release rate decreased, apoptosis decreased, the expression of p38MAPK and BAX decreased, and the expression of BCL-2 increased (all P <0.01). Conclusion MK801 and SB203580 have a protective effect on NMDA-induced cortical neuron injury. The p38MAPK pathway may be involved in the protection of MK801. The common mechanism is through the inhibition of p38MAPK signal pathway-mediated apoptosis-related proteins.