摘要
目的研究五味子醇甲(SA)对过氧化氢(H_(2)O_(2))诱导氧化应激损伤肾小管上皮细胞(HK-2)的保护作用及其机制。方法体外培养HK-2细胞,分为空白对照组,模型对照组(以600μmol·L^(-1)H_(2)O_(2)作用2 h造模)和SA小、中、大剂量组(分别以0.125、0.50和0.75μmol·L^(-1)SA孵育24 h,600μmol·L^(-1)H_(2)O_(2)作用2 h)。CCK-8法检测细胞存活率,抗凝血蛋白膜粘连蛋白-荧光素(Annexin V-FITC/PI)双染法检测细胞凋亡率,碘化丙啶(PI)染色法检测细胞周期,试剂盒检测活性氧(ROS)、超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)、乳酸脱氢酶(LDH)活性,Western Blot测定凋亡通路相关蛋白活性。结果与空白对照组比较,模型对照组细胞存活率显著降低、凋亡率显著上升(P<0.01),G_(0)/G_(1)期染色细胞数量显著升高、S期比例降低(P<0.05),SOD水平与GSH含量显著下降(P<0.01),ROS含量、LDH水平和MDA含量显著升高(P<0.01)。与模型对照组比较,SA小、中、大剂量组细胞凋亡率显著降低(P<0.01);SA中、大剂量组G_(0)/G_(1)期细胞比例明显下降(P<0.05),SA大剂量组S期细胞比例显著升高(P<0.05);SA中、大剂量组ROS水平明显下降(P<0.01),且呈现剂量依赖性;SA小、中、大剂量组SOD活性均上升,但差异无统计学意义;SA中、大剂量组LDH活力下降明显(P<0.01),且与SA浓度呈正相关;SA小、中、大剂量组GSH含量显著上升(P<0.01),且呈现浓度依赖性;SA小、中、大剂量组MDA含量显著下降(P<0.01),且呈现剂量依赖性;SA小、中、大剂量组Bax/Bcl-2比值明显下降(P<0.05),SA中、大剂量组Caspase 3和SA小、中、大剂量组Cytochrome C蛋白活性降低(P<0.05)。结论SA能够调节细胞氧化因子水平,减少细胞凋亡,调控细胞周期进程,对H_(2)O_(2)诱导损伤的HK-2细胞起保护作用。
Objective To investigate the protective effect and mechanism of schisandrin A(SA)on hydrogen peroxide(H_(2)O_(2))-induced oxidative stress injury in renal tubular epithelial cells(HK-2).Methods HK-2 cells were cultured in vitro and divided into five groups:blank control group,model control group(treated with 600μmol·L^(-1) H_(2)O_(2) for 2 h),low-dose SA group(0.125μmol·L^(-1) SA,24 h pretreatment+H_(2)O_(2)),medium-dose SA group(0.5μmol·L^(-1) SA,24 h pretreatment+H_(2)O_(2)),high-dose SA group(0.75μmol·L^(-1) SA,24 h pretreatment+H_(2)O_(2)).The cell survival was assessed by CCK-8 assay;apoptosis level was tested by Annexin V-FITC/PI double staining;cell cycle distribution was detected by propidium iodide staining;oxidative markers(ROS,SOD,MDA,GSH,LDH)was determined with commercial kits;and apoptosis-related proteins(Bax,Bcl-2,Caspase-3,Cytochrome C)was evaluated by Western blot.Results Compared with the blank control,the model group reduced in cell viability and increased in apoptosis(P<0.01),elevated in the ratio of G_(0)/G_(1)-phase cells and decreased in S-phase cells(P<0.05),decreased in SOD activity and GSH content(P<0.01),and increased in the levels of ROS,LDH,and MDA levels(P<0.01).In all SA dose,cell apoptosis reduced(P<0.01).In medium/high dose groups,the G_(0)/G_(1)-phase arrest reduced(P<0.05).In high dose group,the S-phase cells ratio increased(P<0.05).And in medium/high dose groups,ROS(P<0.01)decreased in a dose-dependent manner.The SOD activity increased non-significantly in all SA groups.In SA medium/high dose groups,the LDH activity decreased in a dose-dependent manner.In all SA groups,GSH increased(P<0.01)and MDA decreased,both in a dose-dependent manner.The Bax/Bcl-2 ratio significantly decreased(P<0.05)in all SA groups.The caspase-3 activity decreased in medium/high dose group(P<0.05);and Cytochrome C reduced in all SA groups(P<0.05).Conclusion SA protects HK-2 cells against H_(2)O_(2)-induced oxidative injury by modulating oxidative stress,inhibiting apoptosis,and ameliorating cell cycle arrest.
作者
赵子平
王依
张楠淇
李婉莹
宋明杰
王英平
ZHAO Ziping;WANG Yi;ZHANG Nanqi;LI Wanying;SONG Mingjie;WANG Yingping(College of Chinese Materia Medical,Jilin Agricultural University,National Joint Engineering Research Center for Ginseng Breeding and Development,Changchun 130118,China)
出处
《医药导报》
北大核心
2025年第7期1021-1027,共7页
Herald of Medicine
基金
国家自然科学基金青年基金资助项目(81803733)
吉林省科技发展计划项目医药健康产业发展专项(20210401112YY)。
关键词
五味子醇甲
过氧化氢
急性肾损伤
Schisandrol A
Hydrogen peroxide
Acute kidney injury
