人参皂苷Rg1对过氧化损伤人脐带间充质干细胞的保护作用及机制  

Protective effect of ginsenoside Rg1 on human umbilical cord mesenchymal stem cells damaged by peroxidation

作　　者：种宗雷[1] 杨晓倩[1] 肖以磊[1] 吕甲涛 侯磊[1] 许鹏 朱伟杰[2] CHONG Zonglei;YANG Xiaoqian;XIAO Yilei;LYU Jiatao;HOU Lei;XU Peng;ZHU Weijie(Liaocheng People＇ s Hospital,Liaocheng 252000,China)

机构地区：[1]聊城市人民医院脑科医院,山东聊城252000 [2]济南军区总医院

出　　处：《山东医药》2018年第29期29-32,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81701159);中国博士后基金资助项目(2018M632123);山东省自然科学基金资助项目(ZR2015HQ023);山东省博士后基金资助项目(201703084);山东省重点研发计划项目(2018GSF118046)

摘　　要：目的探讨Rg1对过氧化损伤人脐带间充质干细胞(h UCMSCs)的保护作用及可能的作用机制。方法将h UCMSCs分为6组:阴性对照组,模型组(加入1.6μmol/L H_2O_2),Rg1低、中、高剂量组(模型组条件下培养30min后,加入Rg1各0.5、4、32μmol/L),阳性对照组(模型组条件下培养30 min后,加入220μmol/L维生素C)。采用CCK-8比色法检测细胞增殖活性,ELISA法检测培养上清TNF-α水平,β-半乳糖苷酶染色法检测衰老细胞百分比,免疫组化法检测h UCMSCs中p-p38蛋白表达,Western blotting法检测h UCMSCs中磷酸化c-Jun氨基末端激酶(p-JNK)蛋白表达。结果与模型组相比,随着剂量增加,Rg1各剂量组h UCMSCs增殖活性增加,衰老h UCMSCs百分比减少,TNF-α、p-JNK、p-p38表达降低(P均<0.05)。阳性对照组上述指标介于Rg1高及中、低剂量组之间,差异有统计学意义(P均<0.05)。与阴性对照组比较,模型组,Rg1低、中、高剂量组及阳性对照组h UCMSCs增殖活性降低,衰老h UCMSCs百分比增加,p-JNK、p-p38蛋白及TNF-α表达增加(P均<0.05)。结论 Rg1可能通过抑制炎症反应,降低p-JNK、p-p38表达,从而对过氧化损伤的h UCMSCs产生保护作用。Objective To investigate the protective effect of Rg1 and its possible mechanism on human umbilical cord mesenchymal stem cells（h UCMSCs） damaged by peroxidation. Methods The h UCMSCs were divided into 6 groups： the negative control group,model group（1. 6 μmol/L H2O2）,low-dose,medium-dose,and high-dose Rg1 groups（At 30 min after the same culture method as the model group,0. 5,4,and 32 μmol/L Rg1 were added,respectively）,positive control group（At 30 min after the same culture method as the model group,220 μmol/L vitamin C was added）. The cell proliferation activity was detected by CCK-8 colorimetric assay. The TNF-α level was detected by ELISA. The percentage of senescent cells was detected by β-galactosidase staining. The expression of p-p38 protein in h UCMSCs was detected by immunohistochemistry,and the expression of phosphorylated c-Jun amino terminal kinase（p-JNK） protein in h UCMSCs was detected by Western blotting. Results Compared with the model group,with the increase of dose,the proliferation activity of h UCMSCs increased,the percentage of senescent h UCMSCs decreased,and the expression of TNF-α,p-JNK and p-p38 decreased（all P 〈 0. 05）. The above mentioned indicators in the positive control group were between the high-dose Rg1 and the medium-dose,low-dose groups,and the difference was statistically significant（all P 〈 0. 05）. Compared with the negative control group,the proliferative activity of h UCMSCs in the model group,low-dose,medium-dose,and high-dose Rg1 groups,and the positive control group decreased,but the percentage of senescent h UCMSCs increased,and the expression of p-JNK,p-p38 protein,and TNF-α increased（all P 〈 0. 05）. Conclusion Rg1 may protect the h UCMSCs from peroxidation injury by inhibiting the inflammatory response and decreasing the expression of p-JNK and p-p38.

关 键 词：过氧化损伤 人脐带间充质干细胞 人参皂苷RG1 细胞增殖 细胞衰老 

分 类 号：Q503[生物学—生物化学]