摘要
目的观察人参皂苷Rb1对H2O2损伤RGC-5细胞的保护作用,并探讨其机制。方法采用MTT方法检测人参皂苷Rb1对H2O2损伤RGC-5的细胞活力的影响,试剂盒方法检测SOD活性、GSH含量,免疫荧光化学染色法检测碘化丙啶(PI)、Caspase-3、Ca2+。结果 A、B、C、D、E组在490 nm处检测细胞活力分别为0.266±0.038、0.299±0.025、0.314±0.014、0.222±0.031、0.391±0.041,A、B、C、E组与D组比较,P均<0.05。空白对照组、人参皂苷Rb1组、H2O2损伤组SOD活性分别为19.88±0.12、17.12±0.32、15.47±0.21,人参皂苷Rb1组、空白对照组分别与H2O2损伤组比较,P均<0.05。空白对照组、人参皂苷Rb1组、H2O2损伤组GSH含量分别为15.67±0.18、14.98±0.25、13.71±0.31,人参皂苷Rb1组、空白对照组分别与H2O2损伤组比较,P均<0.05。空白对照组、人参皂苷Rb1组、H2O2损伤组Caspase-3阳性率分别为12%±0.25%、57%±0.47%、28%±0.18%,PI染色阳性率分别为16%±0.25%、18%±0.45%、32%±0.56%,Ca2+荧光染色阳性率分别为8%±0.11%,31%±1.20%,19%±0.34%,人参皂苷Rb1组与H2O2损伤组比较,P均<0.05。结论人参皂苷Rb1能够拮抗H2O2对RGC-5细胞的氧化损伤,其机制可能为提高SOD活性、GSH含量,降低Caspase-3表达,减少Ca2+水平。
Objective To establish RGC-5 cell damage model induced by H 2 O2 , and to observe the protective effect of different dose of ginsenoside Rb 1 on RGC-5 apoptosis induced by H 2 O2 and find the possible mechanisms of the neuro-protection .Methods MTT was used to detect the effect of different dose of ginsenoside Rb 1 on RGC-5 activity induced by H2 O2 .SOD and GSH were detected by Kit method , and PI, Caspase-3, Ca2＋were detected by immunofluorescence chemi-cal dyeing method.Results A, B, C, D, E groups at 490 nm for detection of cell viability were 0.266 ±0.038, 0.299 ±0.025, 0.314 ±0.014, 0.222 ±0.031, 0.391 ±0.041, A, B, C, E group compared with D group, all P〈0.05.The control group, the experimental group, model group, SOD content were 19.88 ±0.12, 17.12 ±0.32, 15.47 ±0.21, and experimental group , control group were compared with the model group , all P〈0.05.Experimental group and control group, model group, GSH content were 15.67 ±0.18 and 14.98 ±0.25, 13.71 ±0.31, and the experimental group and the control group were compared with the model group , all P〈0.05.Compared with the model group , the experimental groups decreased Caspase-3 expression, suggesting that the 0.1 mM of ginsenoside Rb1 can significantly inhibit the expres-sion of Caspase-3.PI staining cells increased significantly in the model group , RGC-5 was obviously injured by H 2 O2 which lead to cell death .Apoptosis of cells were decreased in experimental group .Ca2＋increased obviously in model group . Ca2＋were significantly decreased in experimental group .Conclusion Ginsenosides Rb1 can prevent RGC-5 death in-duced by H2 O2 .The mechanism may be related to up-regulate SOD, GSH activity and reduce the activity of apoptosis-re-lated proteins Caspase-3 and Ca2＋.
出处
《山东医药》
CAS
2014年第47期5-8,共4页
Shandong Medical Journal
基金
国家自然科学基金资助项目(81102828)
山东省自然科学基金资助项目(ZR2011HM011)
