咪达唑仑通过上调miR-320抑制缺氧/复氧诱导的PC12细胞凋亡  

Midazolam inhibits hypoxia/reoxygenation-induced PC12 cell apoptosis through up-regulating miR-320

作　　者：薄云[1] 潘尉洲[1] 周敏 黄岚 BO Yun;PAN Wei-zhou;ZHOU Min;HUANG Lan(The First People's Hospital of Yunnan Province,The Affiliated Hospital of Kunming University of Science and Technology,Kunming 650030,China;Yunnan Cancer Hospital,Kunming 650118,China;Kunming University of science and Technology,Kunming 650093,China)

机构地区：[1]云南省第一人民医院,昆明理工大学附属医院麻醉科,云南昆明650030 [2]云南省肿瘤医院麻醉科,云南昆明650118 [3]昆明理工大学,云南昆明650093

出　　处：《解剖学研究》2024年第2期170-175,共6页Anatomy Research

基　　金：云南省第一人民医院院级课题(2021LCZXXF-HX02)。

摘　　要：目的 为了明确咪达唑仑在缺血性中风中的作用,探讨咪达唑仑对缺氧/复氧(H/R)诱导的PC12细胞增殖和凋亡的影响和潜在机制。方法 体外培养PC12细胞,分为Control组、H/R组、不同剂量咪达唑仑组、10μmol/L咪达唑仑+anti-miR-NC组、和10μmol/L咪达唑仑+anti-miR-320组;CCK-8法和流式细胞术分别用于检测细胞增殖和凋亡;Western Blot检测蛋白表达;RT-qPCR检测miR-320表达。结果 与Control组相比,H/R组细胞A值(0.43±0.02)、pro-caspase3水平(0.14±0.01)和miR-320表达(0.15±0.01)显著降低(P<0.05),而凋亡率(26.68±0.81)和Cleaved-caspase3水平(0.70±0.05)显著升高(P<0.05)。与H/R组相比,不同剂量(0.1、1.0、10μmol/L)咪达唑仑组细胞A值(0.43±0.02、0.57±0.03、0.93±0.04)、pro-caspase3水平(0.14±0.01、0.30±0.02、0.52±0.03)和miR-320表达(0.15±0.02、0.42±0.03、0.81±0.05)显著升高(P<0.05),而凋亡率(25.65±0.92、21.74±0.56、15.63±0.50)和Cleaved-caspase3水平(0.70±0.06、0.50±0.04、0.23±0.02)显著降低(P<0.05),且呈剂量依赖性。与10μmol/L咪达唑仑+anti-miR-NC组相比,10μmol/L咪达唑仑+anti-miR-320组细胞A值(0.47±0.02)、pro-caspase3水平(0.24±0.01)和miR-320表达(0.26±0.02)显著降低(P<0.05),而凋亡率(23.52±0.59)和Cleaved-caspase3水平(0.56±0.04)显著升高(P<0.05)。结论 咪达唑仑可以上调miR-320表达,进而促进H/R诱导的PC12细胞增殖,并抑制凋亡。Objective In order to clarify the role of midazolam in ischemic stroke,the effect of midazolam on hypoxia/reoxygenation(H/R)-induced proliferation and apoptosis of PC12 cells and its potential mechanism were investigated.Methods PC12 cells were cultured in vitro and divided into control group,H/R group,midazolam group with different doses,10μmol/L midazolam+anti-miR-NC group,and 10μmol/L midazolam+anti-miR-320 group.Cell proliferation and apoptosis were detected by CCK-8 method and flow cytometry;Protein expression was tested by Western blot;miR-320 expression was examined by RT-qPCR.Results Compared with the control group,cell A value(0.43±0.02),pro-caspase3 level(0.14±0.01)and miR-320 expression(0.15±0.01)were signif-icantly decreased(P<0.05),while apoptosis rate(26.68±0.81)and Cleaved-caspase3 level(0.70±0.05)were sig-nificantly increased in the H/R group(P<0.05).Compared with the H/R group,cell A value(0.43±0.02,0.57±0.03,0.93±0.04),pro-caspase3 level(0.14±0.01,0.30±0.02,0.52±0.03)and miR-320 expression(0.15±0.02,0.42±0.03,0.81±0.05)were remarkably enhanced(P<0.05),while apoptosis rate(25.65±0.92,21.74±0.56,15.63±0.50)and cleaved-caspase3 level(0.70±0.06,0.50±0.04,0.23±0.02)were significantly reduced in the mid-azolam group with different doses(0.1,1.0,10μmol/L)(P<0.05),in a dose-dependent manner.Compared with the 10μmol/L midazolam+anti-miR-NC group,cell A value(0.47±0.02),pro-caspase3 level(0.24±0.01)and miR-320 expression(0.26±0.02)were markedly inhibited(P<0.05),while apoptosis rate(23.52±0.59)and Cleaved-caspase3 level(0.56±0.04)were remarkably promoted in the 10μmol/L midazolam+anti-miR-320 group(P<0.05).Conclusion Midazolam may upregulate miR-320 expression to promote H/R-induced PC12 cell proliferation and in-hibit apoptosis.

关 键 词：咪达唑仑 PC12细胞 缺氧复氧 miR-320 细胞凋亡 

分 类 号：R96[医药卫生—药理学]