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  

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

作　　者：张素冰[1] 高辉[2] 赵滨滨[1] 张丹琦[1] ZHANG Subing;GAO Hui;ZHAO Binbin;ZHANG Danqi(Department of Anesthesiology and Surgery,The First Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学附属第一医院麻醉手术科,黑龙江哈尔滨150040 [2]黑龙江中医药大学附属第三医院麻醉手术科,黑龙江哈尔滨150000

出　　处：《西部医学》2023年第6期817-821,829,共6页Medical Journal of West China

基　　金：黑龙江省中医药科研项目(ZHY2020-129)。

摘　　要：目的探讨普鲁卡因(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。Objective 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.

关 键 词：普鲁卡因 神经元 miR-138 凋亡 

分 类 号：R743.3[医药卫生—神经病学与精神病学]