丙泊酚联合咪唑安定对脑缺血再灌注PC12细胞模型的保护作用  

Protective effect of propofol combined with midazolam on PC12 cells in cerebral ischemia-reperfusion model

作　　者：李晶金[1] 史艳华[1] 肖黎波[1] 李济成 朱小玲[1] Li Jingjin;Shi Yanhua;Xiao Libo;Li Jicheng;Zhu Xiaoling(Department of Anesthesiology,The First People’s Hospital of Chenzhou,Chenzhou 423000,China)

机构地区：[1]郴州市第一人民医院麻醉科,郴州423000

出　　处：《解剖学杂志》2022年第4期340-346,373,共8页Chinese Journal of Anatomy

摘　　要：目的:探讨丙泊酚(PPF)联合咪唑安定(MID)对糖氧剥夺模型PC12细胞发挥保护作用的分子机制。方法:采用无糖培养基和无氧环境(5%CO_(2)和95%N2)处理PC12细胞构建糖氧剥夺模型,分别用PPF、MID或PPF+MID处理糖氧剥夺模型细胞,或在细胞中转染miR-103-3pmimics、miR-103-3pinhibitor、si-前列腺素内过氧化物合成酶2(PTGS2)、pcDNA-PTGS2或miR-103-3p mimics+pcDNA-PTGS2,采用CCK-8检测细胞增殖活力,流式细胞术检测细胞凋亡率,Fura-2/AM荧光法检测细胞中Ca^(2+)水平,RT-qPCR检测细胞中miR-103-3p水平,免疫印迹检测凋亡相关蛋白(Bcl-2、Bax、c-caspase-3、c-caspase-9)及PTGS2表达水平,双荧光素酶报告基因验证miR-103-3p与PTGS2的靶向关系。结果:PPF联合MID增加糖氧剥夺模型PC12细胞增殖活力,抑制细胞凋亡,并下调模型细胞中Ca^(2+)水平;此外,PPF联合MID处理糖氧剥夺模型PC12细胞,上调细胞中miR-103-3p水平;miR-103-3p可靶向下调PC12细胞中PTGS2的蛋白水平;过表达miR-103-3p与沉默PTGS2均可缓解糖氧剥夺诱导对PC12细胞造成的损伤,敲降miR-103-3p与过表达PTGS2则加剧糖氧剥夺模型下PC12的损伤。结论:PPF联合MID通过上调miR-103-3p抑制PTGS2表达缓解糖氧剥夺模型中PC12细胞损伤。Objective:To investigate the molecular mechanism of protective effect of propofol(PPF) combined with midazolam(MID) on PC12 cells in constructed oxygen glucose deprivation(OGD)model.Methods:PC12cells were treated with glucose free medium and an aerobic environment(5% CO_(2) and 95% N2) to construct OGD model.OGD model cells were treated with PPF,MID or PPF+MID;besides,miR-103-3p mimics,miR-103-3p inhibitor,si-prostaglandin-endoperoxide synthase 2(PTGS2),pcDNA-PTGS2 or miR-103-3p mimics+pcDNAPTGS2 were transfected into PC12.Cell proliferation was detected by CCK-8;apoptosis rate was measured by flow cytometry.Ca^(2+) level was detected by Fura-2/AM fluorescence assay;expression of miR-103-3p was measured by RT-qPCR.Expression of apoptosis related proteins(Bcl-2,Bax,c-caspase-3 and c-caspase-9),and PTSG2 were detected by Western blotting.The relationship between miR-103-3p and PTGS2 was confirmed by dual-luciferase reporter gene assay system.Results:PPF combined with MID increased the proliferation,decreased apoptosis and Ca^(2+) level of PC12 in OGD model.Additionally,it also markedly up-regulated the expression of miR-103-3p in PC12.Moreover,miR-103-3p target down-regulated the protein level of PTGS2 in PC12.Overexpression of miR-103-3p and silencing of PTGS2 alleviated the injury of PC12 cells induced by OGD,and knockdown of miR-103-3p and overexpression of PTGS2 aggravated the injury of PC12 in OGD model.Conclusion:PPF combined with MID alleviates injury of PC12 cell in OGD model by up-regulating miR-103-3p and inhibiting PTGS2.

关 键 词：丙泊酚 咪唑安定 糖氧剥夺 PC12 增殖 凋亡 CA^(2+) 

分 类 号：R96[医药卫生—药理学]