曲美他嗪通过上调Bcl-2改善骨髓间质干细胞存活

中国临床药理学与治疗学 ›› 2011, Vol. 16 ›› Issue (7): 729-735.

曲美他嗪通过上调Bcl-2改善骨髓间质干细胞存活

朱刚艳¹, 徐红新², 田毅浩³, 马红梅¹, 唐世琪¹, 汪福良¹, 邬松林¹

¹武汉大学人民医院老年病科,
²心内科,武汉 430060,湖北;
³武汉大学基础医学院解剖教研室,武汉 430071,湖北

收稿日期:2011-04-25 修回日期:2011-06-07 出版日期:2011-07-26 发布日期:2011-09-22
作者简介:朱刚艳,女,博士,主任医师,研究方向:老年病学。Tel: 13607129038 E-mail: zhugy99@yahoo.com.cn
基金资助:
国家自然科学基金资助课题(30700314);湖北省武汉市科技局课题(20065004116-02)

Ameliorative effects of trimetazidine on bone marrow mesenchymal stem cells viability through Bcl-2 expression

ZHU Gang-yan¹, XU Hong-xin², TIAN Yi-hao³, MA Hong-mei¹, TANG Shi-qi¹, WANG Fu-liang¹, WU Song-lin¹

¹Department of Geriatrics, Renmin Hospital of Wuhan University,
²Department of Cardiology, Wuhan 430060, Hubei,China;
³Department of Anatomy, Basal Medical College of Wuhan University, Wuhan 430071, Hubei,China

Received:2011-04-25 Revised:2011-06-07 Online:2011-07-26 Published:2011-09-22

摘要/Abstract

摘要： 目的: 探讨曲美他嗪(trimetazidine, TMZ)能否改善骨髓间质干细胞(bone marrow mesenchymal stem cells, MSCs)在体外缺氧模型及急性心肌梗死(acute myocardial infarction, AMI)大鼠心脏的存活和分化,并讨论其机制。方法: 加入或未加入曲美他嗪的MSCs在无血清培养基培养并缺氧暴露 12 h,采用透射电子显微镜和流式细胞仪检查第3代MSCs的活力和凋亡。30只Wistar大鼠随机分为3组(各组n=10):AMI对照组,单纯MSCs移植组,MSCs移植+曲美他嗪组。结扎左冠状动脉前降支制备AMI模型。将MSCs(细胞数4×10⁷)注入梗死心肌边缘(MSCs组和MSCs+曲美他嗪组)。曲美他嗪+MSCs组大鼠在AMI前 3 d 开始至AMI后 28 d 加喂曲美他嗪,剂量 2.08 mg·kg^-1·d^-1。对照组动物注射无血清培养基。移植 28 d 后,超声心动图评估心脏结构和功能。免疫荧光染色检测移植细胞在体内的存活和分化。TUNEL法检测梗死周围区域的细胞凋亡。通过免疫印迹检测梗死后心肌中的细胞凋亡相关蛋白(Bcl-2和Bax)。结果: 缺氧培养下,曲美他嗪处理过的MSCs细胞凋亡降低了一半。在体内,与对照组相比,MSCs组和MSCs+曲美他嗪组的心肌梗死面积显著缩小,心功能明显改善。与单纯MSCs移植相比,曲美他嗪与MSCs移植的组合治疗表现出了更低的干细胞凋亡,更高的干细胞存活,更小的心肌梗死面积,与进一步改善的心功能。免疫印迹法表明,与MSCs组相比,在曲美他嗪+MSCs组中,抗凋亡蛋白Bcl-2表达上调,而促凋亡蛋白Bax蛋白下调。结论: 联合曲美他嗪的MSCs移植治疗,在MSCs的存活和心脏功能的恢复方面优于单纯MSCs移植,其机制可能是通过上调抗凋亡蛋白Bcl-2表达。

关键词: 曲美他嗪, 骨髓间质干细胞, 移植, 心肌梗死

Abstract: AIM: To investigate whether Trimetazidine (TMZ) can improve the survival of Bone marrow mesenchymal stem cells (MSCs) in an ex-vitro model of hypoxia, as well as survival, differentiation, and subsequent activities of transplanted MSCs in rat hearts with acute myocardial infarction (AMI), and discuss the mechanism.METHODS: MSCs at passage 3 were examined for their viability and apoptosis by transmission electron microscope, and flow cytometry following cultured in serum-free medium and exposed to hypoxia (5%CO₂, 95%N₂) for 12 h with or without TMZ. Thirty wistar rats were divided into 3 groups (n=10 in every group), including group I (AMI control), group II (MSCs transplantation alone), and group III (TMZ+MSCs). Rat MSCs (4×10⁷cells) were injected into peri-infarct myocardium (MSCs and TMZ+MSCs groups) thirty minutes after coronary artery ligation. The rats in TMZ+MSCs group were additionally fed TMZ with 2.08 mg·kg^-1·d^-1 from 3 days before AMI to 28 days after AMI. Cardiac structure and function was assessed by echocardiography 28 days after transplantation. The survival and differentiation of transplanted cells were detected by immunofluorescent staining. The cellular apoptosis in the peri-infarct region was detected with TUNEL assay. Furthermore, apoptosis-related proteins (Bcl-2, Bax) within the post-infarcted myocardium were detected with Western blotting.RESULTS: In hypoxic culture, the TMZ-treated MSCs displayed a two-fold decrease in apoptosis under serum-free medium and hypoxia environment. In vivo, cardiac infarct size was significantly smaller, cardiac function significantly improved in the MSCs and TMZ+MSCs groups than those in the control group. Combined treatment with TMZ and MSCs implantation demonstrated a further decrease in MSCs apoptosis, a further increase in MSCs viability, a further decrease in infarct size, and a further improvement in cardiac function as compared with MSCs alone. Western blotting indicated that the expression of anti-apoptotic protein Bcl-2 was upregulated, while the pro-apoptotic protein Bax was downregulated in the TMZ+MSCs group, compared with that in the MSCs group.CONCLUSION: Implantation of MSCs combined with TMZ treatment is superior to MSCs monotherapy for MSCs viability and cardiac function recovery. The up-regulating of Bcl-2 maybe the potential mechanism.

Key words: Trimetazidine, Bone marrow mesenchymal stem cells, Implantation, Myocardial infarction

中图分类号:

R965.2

朱刚艳, 徐红新, 田毅浩, 马红梅, 唐世琪, 汪福良, 邬松林. 曲美他嗪通过上调Bcl-2改善骨髓间质干细胞存活[J]. 中国临床药理学与治疗学, 2011, 16(7): 729-735.

ZHU Gang-yan, XU Hong-xin, TIAN Yi-hao, MA Hong-mei, TANG Shi-qi, WANG Fu-liang, WU Song-lin. Ameliorative effects of trimetazidine on bone marrow mesenchymal stem cells viability through Bcl-2 expression[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2011, 16(7): 729-735.

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