人参皂苷Rb3通过抗氧化应激对HT22细胞氧糖剥夺-复糖复氧损伤的保护作用  

Protective Effect of Ginsenoside Rb3 on OGD-R Injury of HT22 Cells by Anti-oxidative Stress

作　　者：罗苑 陈普 段小花 李付惠[1] 冯光泉[1] 常征[1] 高明菊[1] 詹云静[1] 李建平[1] 王朝勇[1] LUO Yuan;CHEN Pu;DUAN Xiao-hua;LI Fu-hui;FENG Guang-quan;CHANG Zheng;GAO Ming-ju;ZHAN Yun-jing;LI Jian-ping;WANG Chao-yong(School of Notoginseng Medicine,Wenshan University,Wenshan 663099,China;Yunnan Key Laboratory of Dai Medicine and Yi Medicine,College of Ethnic Medicine,Yunnan University of Chinese Medicine,Kunming 650500,China)

机构地区：[1]文山学院三七医药学院,云南文山663099 [2]云南中医药大学民族医药学院,云南省傣医药与彝医药重点实验室,云南昆明650500

出　　处：《中国兽医杂志》2022年第9期74-79,87,共7页Chinese Journal of Veterinary Medicine

基　　金：云南省教育厅科学研究基金(2020J0710)。

摘　　要：为了探讨人参皂苷Rb3对氧糖剥夺-复糖复氧(OGD/R)损伤的HT22细胞氧化应激的抑制作用和神经保护作用,本试验将体外培养的HT22细胞随机分为对照组(Control group)、模型组(OGD/R group)、阳性组(EDA group,OGD/R+EDA 100μmol/L)和Rb3组(Rb3 group,OGD/R+Rb32.5、5、10μmol/L);用连二亚硫酸钠(Na_(2)S_(2)O_(4))10 mmol/L合并无糖DMEM培养基进行氧糖剥夺培养2 h,后复糖复氧2 h以建立体外OGD/R模型,在光学显微镜下观察细胞形态;继而用噻唑蓝(MTT)和乳酸脱氢酶(LDH)检测细胞存亡率,并用试剂盒检测氧化应激相关指标。结果显示,与对照组相比,模型组细胞肿胀、突起消失,死亡率极显著升高(P<0.01),氧化应激相关指标活性氧(ROS)、一氧化氮(NO)水平和丙二醛(MDA)含量极显著增加(P<0.01),抗氧化酶超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性极显著下降(P<0.01),细胞色素C(Cyt-C)释放量极显著增加(P<0.01);与模型组相比,Rb3组可显著改善细胞形态,并极显著提高细胞存活率和降低死亡率(P<0.01),氧化应激方面可极显著降低ROS、NO水平和MDA含量(P<0.01),极显著提高抗氧化酶SOD、GSH-Px的活性(P<0.01),极显著抑制Cyt-C的释放(P<0.01)。结果表明,人参皂苷Rb3对OGD/R诱导的脑缺血再灌注损伤(CIRI)的体外模型具有明显的保护作用,其机制可能与抗氧化应激、抑制Cyt-C的释放以减少内源性凋亡有关。In order to investigate the inhibitory oxidative stress and neuroprotective effects of ginsenoside Rb3 on HT22 cells injured by oxygen-glucose deprivation/reperfusion(OGD/R)injury,HT22 cells cultured in vitro were randomly divided into control group,model group(OGD/R group),and positive drug group(EDA group,OGD/R+EDA 100μmol/L)and Rb3 group(OGD/R+Rb32.5,5,10μmol/L);the OGD/R model was established in vitro by using 10 mmol/L sodium dithionite(Na_(2)S_(2)O_(4))combined with a sugar-free DMEM medium for 2 h of oxygen-glucose deprivation,followed by re-oxygenation for 2 h,the morphology of the cells was observed under a light microscope;the MTT and lactate dehydrogenase(LDH)were used to detect cell survival rate;and kits were used to detect oxidative stress-related indicators.The results showed that compared with the control group,the cells in the OGD/R group swelled,the protrusion disappeared,and the mortality rate was significantly increased(P<0.01);the oxidative stress-related indicators,reactive oxygen species(ROS),NO levels,and malondialdehyde(MDA)content all significantly increased(P<0.01);both antioxidant enzymes superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)activity were significantly decreased(P<0.01),and cytochrome C(Cyt-C)release was significantly increased(P<0.01).Compared with the OGD/R group,the Rb3 group significantly improved cell morphology,significantly increased cell survival rate and decreased cell mortality(P<0.01);significantly decreased ROS,NO levels and MDA content(P<0.01)in oxidative stress;significantly increased the activities of SOD and GSH-Px(P<0.01)and inhibited the release of Cyt-C(P<0.01).Thus,ginsenoside Rb3 has a significant protective effect on OGD/R induced cerebral ischemia reperfusion injury(CIRI)in vitro,possibly through antioxidant stress and inhibition of Cyt-C release to reduce endogenous apoptosis.

关 键 词：人参皂苷RB3 氧糖剥夺-复糖复氧 氧化应激 凋亡 

分 类 号：R29[医药卫生—民族医学]