李宗伟, 付荣, 赵亚蕊, 赵超, 李卓玉*
本研究采用缺氧、血清饥饿(hypoxia and serum deprivation, H/SD)联合处理骨髓间充质干细胞(mesenchymal stem cells, MSCs)，模拟MSCs移植的心脏缺血微环境，研究蛋白酶体抑制剂MG-132对H/SD诱导的MSCs凋亡和旁分泌的影响。采用Annexin V/PI流式细胞术检测细胞凋亡，real-time PCR检测白介素-1β (IL-1β)、肿瘤坏死因子-α (TNF-α)和白介素-10 (IL-10) mRNA表达，免疫荧光染色检测NF-κBp65细胞核转位，Western blot检测IL-1β和TNF-α蛋白表达，ELISA检测IL-10细胞分泌。结果显示，MG-132能够抑制H/SD诱导的MSCs凋亡，并通过抑制NF-κBp65细胞核转位抑制IL-1β和TNF-α mRNA转录；MG-132能够明显增强H/SD诱导的IL-10 mRNA转录和IL-10分泌。上述结果提示，MG-132能够抑制缺血诱导的MSCs凋亡和炎症因子IL-1β和TNF-α表达，增强抗炎症因子IL-10的分泌。MG-132预处理有可能成为提高MSCs移植存活率、增强MSCs旁分泌效应的有效策略。
[MG-132 enhances MSCs survival and IL-10 secretion under hypoxia and serum deprivation condition.] [Article in Chinese]
LI Zong-Wei, FU Rong, ZHAO Ya-Rui, ZHAO Chao, LI Zhuo-Yu*
Institute of Biotechnology, Key Laboratory of Chemical Biology and Molecular Engineering of National Ministry of Education, Shanxi University, Taiyuan 030006, China
Bone marrow-derived mesenchymal stem cells (MSCs) have emerged as attractive candidates for cellular therapies for heart and other organ-system disorders. However, a major dilemma in stem cell therapy for ischemic heart diseases is the low survival of transplanted cells in the ischemic and peri-infarcted region. In this study, MSCs were treated by hypoxia and serum deprivation (H/SD) to mimic the ischemic microenvironment of infarcted hearts where MSCs were transplanted. The effects of proteasome inhibitor MG-132 on H/SD-induced apoptosis and paracrine effects were investigated. Apoptosis of MSCs was detected by Annexin V-FITC flow cytometric analysis. Transcriptional levels of IL-1β, TNF-α and IL-10 were examined by real-time PCR. The nuclear translocation of NF-κBp65 was assessed by immunocytochemical staining. Translational changes of IL-1β and TNF-α were detected by Western blot. The secretion of IL-10 from MSCs was examined by ELISA assay. The results showed that MG-132 could effectively suppress H/SD-induced MSCs apoptosis. Furthermore, the induced IL-1β and TNF-α transcription was also inhibited by MG-132 treatment, which may be due to the inhibition of NF-κBp65 nuclear translocation by MG-132. Importantly, MG-132 effectively enhanced H/SD-induced transcription and secretion of IL-10, which is an important paracrine factor from MSCs. Our findings suggest that pretreatment of MSCs by MG-132 before cell transplantation may be an effective strategy to improve cell survival and enhance paracrine effects of MSCs.
通讯作者：李卓玉 E-mail: firstname.lastname@example.org
李宗伟, 付荣, 赵亚蕊, 赵超, 李卓玉. MG-132对缺氧、血清饥饿诱导的MSCs凋亡和旁分泌的影响[J]. 生理学报 2011; 63 (6): 525-532.
LI Zong-Wei, FU Rong, ZHAO Ya-Rui, ZHAO Chao, LI Zhuo-Yu. [MG-132 enhances MSCs survival and IL-10 secretion under hypoxia and serum deprivation condition.] [Article in Chinese]. Acta Physiol Sin 2011; 63 (6): 525-532 (in Chinese with English abstract).