白桦脂酸对H_(2)O_(2)诱导的PC12神经细胞氧化损伤和细胞凋亡影响  

Effect of Betulinic Acid on Oxidative Damage and Apoptosis of PC12 Nerve Cells Induced by H_(2)O_(2)

作　　者：赵晓羲 李英[1] 张家齐 ZHAO Xiaoxi;LI Ying;ZHANG Jiaqi(Second Stroke Ward of Shenyang Second Hospital of Traditional Chinese Medicine,Shenyang,110101,China)

机构地区：[1]沈阳市第二中医医院中风二病区,沈阳市110101

出　　处：《医学分子生物学杂志》2021年第6期428-433,共6页Journal of Medical Molecular Biology

基　　金：辽宁省中医药管理局科研项目(No.LNZYXZK201906)。

摘　　要：目的探讨白桦脂酸对H_(2)O_(2)诱导的PC12神经细胞氧化损伤和凋亡的影响。方法PC12神经细胞分成Control、Model(H_(2)O_(2)刺激)、BA-L(1μmol/L的白桦脂酸和H_(2)O_(2)刺激)、BA-M(2μmol/L的白桦脂酸和H_(2)O_(2)刺激)、BA-H(4μmol/L的白桦脂酸和H_(2)O_(2)刺激)、BA-H+PMA组(1μmol/L的白桦脂酸、H_(2)O_(2)、NF-κB信号通路激活剂PMA刺激),CCK-8方法分析细胞增殖活性变化,流式细胞术分析细胞凋亡水平变化,Western印迹分析细胞中Bax、Bcl-2、P65蛋白表达变化,黄嘌呤氧化法检测SOD水平,比色法检测GSH-Px水平、硫代巴比妥酸法检测MDA水平、CellROX Green荧光法检测ROS水平。结果与Control组比较,Model组PC12神经细胞增殖活性降低,细胞凋亡率升高,细胞中Bax蛋白表达水平升高,Bcl-2蛋白表达水平降低,SOD、GSH-Px活性降低,MDA、ROS水平升高,P65蛋白水平升高。与Model组比较,BA-L、BA-M、BA-H组PC12神经细胞增殖活性逐渐升高,细胞凋亡率逐渐降低,细胞中Bax蛋白表达逐渐减少,Bcl-2蛋白表达逐渐增加,SOD、GSH-Px活性逐渐升高,MDA、ROS水平逐渐降低,P65蛋白水平逐渐降低。与BA-H组比较,BA-H+PMA组C12神经细胞增殖活性降低,细胞凋亡率升高,细胞中Bax、P65蛋白表达增多,Bcl-2蛋白表达减少,SOD、GSH-Px活性降低,MDA、ROS水平升高。结论白桦脂酸通过下调NF-κB信号抑制H_(2)O_(2)诱导的PC12神经细胞氧化损伤和细胞凋亡。Objective To investigate the effect of betulinic acid on the oxidative damage and apoptosis of PC12 nerve cells induced by H_(2)O_(2).Methods PC12 nerve cells were divided into Control,Model(H_(2)O_(2) stimulation),BA-L(1μmol/L betulinic acid and H_(2)O_(2) stimulation),BA-M(2μmol/L betulinic acid and H_(2)O_(2) stimulation),BA-H(4μmol/L betulinic acid and H_(2)O_(2) stimulation),BA-H+PMA group(1μmol/L betulinic acid,H_(2)O_(2),NF-κB signaling pathway activator PMA stimulation),CCK-8 method was used to analyze the changes in cell proliferation activity,and flow cytometry was used to analyze the level of apoptosis changes,Western blot analysied of Bax,Bcl-2,P65 protein expression changes in cells,Xanthine oxidation method to detect SOD level,colorimetric method to detect GSH-Px level,thiobarbituric acid method to detect MDA level,CellROX Green fluorescence method detected the level of ROS.Results Compared with the Control group,the PC12 nerve cell proliferation activity in the Model group decreased,the apoptosis rate increased,the Bax protein expression level in the cells increased,the Bcl-2 protein expression level decreased,SOD,GSH-Px activity decreased,MDA,ROS rised,and the P65 protein level rised.Compared with the Model group,the proliferation activity of PC12 nerve cells in the BA-L,BA-M,and BA-H groups gradually increased,the apoptosis rate gradually decreased,the Bax protein expression in the cells gradually decreased,the Bcl-2 protein expression gradually increased,and the SOD,GSH-Px activity gradually increased,MDA and ROS levels gradually decreased,and P65 protein levels gradually decreased.Compared with the BA-H group,the BA-H+PMA group C12 nerve cell proliferation activity decreased,the apoptosis rate increased,the expression of Bax and P65 protein in the cell increased,the expression of Bcl-2 protein decreased,and the activity of SOD and GSH-Px decreased,MDA and ROS levels increased.Conclusion Betulinic acid inhibits the oxidative damage and apoptosis of PC12 nerve cells induced by H_(2)O_(2).

关 键 词：白桦脂酸 PC12神经细胞 H_(2)O_(2) 氧化损伤 凋亡 

分 类 号：R285[医药卫生—中药学]