山楂叶总黄酮通过调控miR-133b改善缺氧复氧诱导的神经细胞损伤  

Effects of hawthorn leaf flavonoids on improvement of hypoxia-reoxygenation-induced neuronal damage via miR-133b regulation

作　　者：李丽静[1] 王领弟[1] 吴晓光 LI Li-jing;WANG Ling-di;WU Xiao-guang(Department of Pharmacy,The Hospital Affiliated to Chengde Medical College,Chengde 067000,China;Hebei Provincial Key Laboratory for Development and Research of Traditional Chinese Medicine,Chengde 067000,China)

机构地区：[1]承德医学院附属医院药学部,河北承德067000 [2]河北省中药开发与研究重点实验室,河北承德067000

出　　处：《中成药》2020年第6期1443-1449,共7页Chinese Traditional Patent Medicine

基　　金：河北省高等学校科学研究课题计划(QN2014103)。

摘　　要：目的探讨山楂叶总黄酮通过调控miR-133b改善缺氧复氧诱导的神经细胞损伤。方法山楂叶总黄酮(10、30、90 mg/L)培养SH-SY5Y细胞24 h后,构建缺氧复氧(H/R)细胞模型;将miR-NC、miR-133b、anti-miR-NC、anti-miR-133b质粒转染至SH-SY5Y细胞6 h,加入90 mg/L山楂叶总黄酮处理24 h,再进行缺氧复氧处理。RT-PCR检测miR-133b、RBMX mRNA表达,Western blot检测RBMX、Bax、Bcl-2蛋白表达,采用试剂盒分别检测LDH、SOD、MDA水平,流式细胞术检测细胞凋亡,双荧光素酶报告基因实验检测荧光活性。结果预处理SH-SY5Y细胞,山楂叶总黄酮能上调H/R诱导的神经细胞SOD水平,下调LDH、MDA水平,抑制Bax、RBMX表达及细胞凋亡率,促进Bcl-2、miR-133b表达(P<0.05),与H/R诱导的神经细胞过表达miR-133b结果一致。且miR-133b靶向调控RBMX的表达,抑制miR-133b表达逆转了山楂叶总黄酮对H/R诱导的神经细胞损伤的改善作用。结论山楂叶总黄酮通过调控miR-133b靶向调控RBMX的表达改善缺氧复氧诱导的神经细胞损伤。AIM To investigate the effects of hawthorn leaf flavonoids(HLF)on improvement of hypoxia-reoxygenation-induced neuronal damage via miR-133 b regulation.METHODS HLF(10, 30, 90 mg/L) cultured SH-SY5 Y cells for 24 h were induced into hypoxia-reoxygenation(H/R) cell models. The SH-SY5 Y cells were then transfected with miR-NC, miR-133 b, anti-miR-NC, or anti-miR-133 b plasmids for 6 h, respectively, and treated with 90 mg/L HLF for 24 h, followed by H/R. Their detections of expression of miR-133 b and RBMX mRNA by RT-PCR, expression of RBMX, Bax, and Bcl-2 protein by Western blot were performed. The levels of LDH, SOD, and MDA were determined by the kit. The determinations of cell apoptosis by flow cytometry, fluorescent activity by dual luciferase reporter gene experiment were carried out as well.RESULTS SH-SY5 Y cells pretreated with HLF shared up-regulated H/R-induced neuronal SOD levels, down-regulated LDH, MDA levels, inhibited Bax, RBMX expression and apoptosis rate, promoted Bcl-2, miR-133 b expression(P<0.05), which was consistent with the results of mir-133 b overexpression in H/R-induced nerve cells. Moreover, miR-133 b targeted the regulation of RBMX expression, and inhibition of miR-133 b expression reversed the protective effect of hawthorn leaf flavonoids on neuronal damage induced by H/R.CONCLUSION HLF can improve H/R-induced neuronal damage by regulating the expression of RBMX via miR-133 b regulation.

关 键 词：山楂叶总黄酮 miR-133b RBMX 神经细胞损伤 缺氧复氧 

分 类 号：R285􀆰[医药卫生—中药学]