人参皂苷Rg1对缺氧复氧BMSCs增殖和凋亡的影响

doi:10.13418/j.issn.1001-165x.2015.06.009

中国临床解剖学杂志 ›› 2015, Vol. 33 ›› Issue (6): 655-661.doi: 10.13418/j.issn.1001-165x.2015.06.009

人参皂苷Rg1对缺氧复氧BMSCs增殖和凋亡的影响

王艳¹，　闵鹤鸣²，　张苗苗³，　秦书俭³，　闵连秋¹，　包翠芬⁴

1. 辽宁医学院附属第一医院神经内科； 2. 辽宁医学院细胞生物学教研室； 3.辽宁医学院解剖学教研室；
4.辽宁省高校分子生物与新药开发重点实验室，辽宁锦州 121000

收稿日期:2015-06-10 出版日期:2015-11-25 发布日期:2015-12-18
通讯作者: 包翠芬，教授，硕士生导师，E-mail: mianyizuhua@ aliyun.com
作者简介:王艳（1988-），女，河南信阳人，硕士，主要研究方向为脑血管疾病防治研究，E-mail：wangyan198811152@163.com
基金资助:
国家自然基金（81202783,31170930）

Effects of ginsenoside Rg1 on BMSCs proliferation and apoptosis after hypoxia-reoxygenation

WANG Yan¹, MIN He-ming², ZHANG Miao-miao³,QIN Shu-jian³， MIN Lian-qiu¹，BAO Cui-fen⁴

1. Department of Neurology, the First Affiliated Hospital，Liaoning Medical University, Jinzhou 121001, China； 2. Department of Cell Biology，Liaoning Medical University, Jinzhou 121001, China； 3. Department of Anatomy，Liaoning Medical University, Jinzhou 121001, China； 4. Key lab of Molecular Cell Biology and New Drug Development, Liaoning Medical University, Jinzhou 121001, China

Received:2015-06-10 Online:2015-11-25 Published:2015-12-18

摘要/Abstract

摘要：

目的　探讨人参皂苷Rg1(ginsenoside Rg1)对缺氧复氧BMSCs增殖和凋亡的影响,并探讨其可能机制。方法　实验分为BMSCs正常对照组、BMSCs缺氧复氧组(Model组)、人参皂苷Rg1 1×10-7、1×10-6 、1×10-5 mol/L处理组、尼莫地平2.5×10-7 mol/L处理组（阳性对照组）。各组分别于缺氧复氧12h前加入完全培养基（正常对照组、Model组）、人参皂苷Rg1（人参皂苷Rg1各处理组）、尼莫地平（阳性对照组）。建立缺氧复氧BMSCs模型。采用TUNEL法检测各组的细胞凋亡率，免疫荧光和免疫印迹技术定性定量检测各组增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、Bcl-2、Bax的表达情况。结果 TUNEL结果显示，BMSCs正常对照组未见明显凋亡细胞，Model组可见明显的凋亡细胞，与Model组比较，其余各处理组凋亡细胞数量均减少，以人参皂苷Rg1（1×10-5mol/L）组最为明显。免疫荧光和免疫印迹结果显示，BMSCs正常对照组可见少量的PCNA 、bcl-2、bax表达；Model组的PCNA 、bcl-2的表达减少，但bax的表达显著增高，bcl-2/bax比值降低；与Model组比较，人参皂苷Rg1各组PCNA 、bcl-2表达均呈不同程度的增高，而bax表达呈现相反的趋势，bcl-2/bax比值增高，以Rg1 (1×10-5 mol/L)组最为明显。结论人参皂苷Rg1预处理对缺氧复氧BMSCs具有保护作用，其机制可能与下调bax的表达，上调PCNA 、bcl-2的表达，抑制BMSCs凋亡和促进BMSCs增殖有关。

关键词: 人参皂苷Rg1, BMSCs, 增殖, 凋亡

Abstract:

Objective　To investigate the effects of ginsenoside Rg1 on BMSCs proliferation and apoptosis after acute hypoxia-reoxygenation（H/R）and the possible mechanism.　Methods　BMSCs were divided into normal control group，model group，ginsenoside Rg1 1×10-7，1×10-6 and 1×10-5 mol/L groups, Nimodipine 2.5×10-7 mol/L group（positive control group）. In the 12 hours before building H/R model，every groups were respectively added normal saline (normal control group, model group), ginsenoside Rg1 (ginsenoside Rg1 treated groups), Nimodipine ( positive control group). Acute H/R BMSCs model was established. TUNEL was used to count apoptosis rate of cell and IF and Western blot technique was used to observe the expression of PCNA、bcl-2、bax qualitatively and quantitatively. Results Normal control group showed no significant apoptotic cells，model group showed obvious apoptotic cells，compared with model group, the number of apoptotic cells of every groups decreased, Rg1 (1×10-5 mol/L) group was the most obvious. Compared with normal control group, the expression of PCNA and bcl-2 of model group decreased,but the expression of bax significantly increased, and the ratio of bcl-2/bax decreased; Compared with model group，the expression of PCNA and bcl-2 of ginsenoside Rg1 treated groups increased in a different degree,but the expression of bax showed the opposite trend; the ratio of bcl-2/bax increased , and ginsenoside Rg1 (1×10-5 mol/L) group was the most obvious. Conclusion Ginsenoside Rg1 can protect BMSCs after acute anoxia-reoxygenation by down-regulating the expression of bax and up-regulating the expression of PCNA and bcl-2 and by inhibiting apoptosis and promoting proliferation.

Key words: Ginsenoside Rg1, BMSCs, Proliferation, Apoptosis

王艳,闵鹤鸣,张苗苗,秦书俭,闵连秋,包翠芬. 人参皂苷Rg1对缺氧复氧BMSCs增殖和凋亡的影响[J]. 中国临床解剖学杂志, 2015, 33(6): 655-661.

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人参皂苷Rg1对缺氧复氧BMSCs增殖和凋亡的影响

Effects of ginsenoside Rg1 on BMSCs proliferation and apoptosis after hypoxia-reoxygenation

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