普鲁卡因通过调控miR-138抑制OGD诱导的大鼠皮层神经元损伤

探讨普鲁卡因（PROC）对氧糖剥夺（OGD）诱导大鼠皮层神经元损伤的影响及可能机制。方法 体外培养大鼠皮层神经元，用不同剂量（0.00、3.50、7.00、14.00 mg/L）PROC干预大鼠皮层神经元24 h，建立OGD模型。CCK-8法检测细胞存活率，试剂盒检测乳酸脱氢酶（LDH）漏出率，流式细胞术检测细胞凋亡，蛋白质印迹法检测活化的半胱天冬酶（cleaved-caspase）3和cleaved-caspase9蛋白表达，qRT-PCR检测细胞中miR-138表达。转染miR-138模拟物或抑制剂至大鼠皮层神经元细胞后，建立OGD模型，上述相同方法检测神经元损伤情况。结果 与对照组相比，OGD组存活率、miR-138表达降低，LDH释放率、凋亡率及cleaved-caspase3和cleaved-caspase9蛋白表达增加（均P＜0.05）。与OGD组比较，OGD+PROC-L组、OGD+PROC-M组和OGD+PROC-H组存活率、miR-138表达增加，LDH释放率、凋亡率及cleaved-caspase3和cleaved-caspase9蛋白表达降低（均P＜0.05）。过表达miR-138后，OGD诱导的大鼠皮层神经元存活率增加，LDH漏出率、凋亡率及cleaved-caspase3和cleaved-caspase9蛋白表达降低（均P＜0.05）。敲减miR-138可逆转PROC对OGD诱导大鼠皮层神经元损伤的影响。结论PROC可能通过上调miR-138，抑制OGD诱导的大鼠皮层神经元损伤

Procaine inhibits oxygen-glucose deprivation-induced rat cortical neurons damage by regulating miR-138

To investigate the effect and possible mechanism of procaine (PROC) on oxygen-glucose deprivation (OGD)-induced rat cortical neurons damage.Methods Rat cortical neurons were cultured in vitro and intervened with different doses (3.50, 7.00, 14.00 mg/L) of PROC for 24 h, and then the OGD model was established. The cell survival rate was detected by CCK-8 method, LDH leakage rate was detected by kit. Flow cytometry was used to detect cell apoptosis, and Western blotting was used to detect the protein expression of cleaved-caspase3 and cleaved-caspase9. qRT-PCR was used to detect the expression of miR-138 in cells. After rat cortical neuron were transfected with miR-138 mimics or inhibitors, the OGD model was established, and neuronal damage was detected by the same method as above.Results Compared with the Con group, the survival rate and the expression of miR-138 in the OGD group decreased,while the LDH leakage rate, apoptosis rate and the protein expressions of cleaved-caspase3 and cleaved-caspase9 increased. Compared with OGD group, OGD+PROC-L group, OGD+PROC-M group and OGD+PROC-H group increased the survival rate, miR-138 expression, LDH leakage rate, apoptosis rate and cleaved-caspase3 and cleaved-caspase9 protein Expression decreased. After overexpression of miR-138, the survival rate of OGD-induced rat cortical neurons increased, while the LDH leakage rate, apoptosis rate, and the protein expressions of cleaved-caspase3 and cleaved-caspase9 decreased. Knockdown of miR-138 reverses the effect of PROC on OGD-induced rat cortical neuron damage. Conclusion PROC may inhibit OGD-induced rat cortical neurons damage by up-regulating miR-138