Heat shock protein 90 protects rat mesenchymal stem cells against hypoxia and serum deprivation-induced apoptosis via the PI3K/Akt and ERK1/2 pathways
Heat shock protein 90 protects rat mesenchymal stem cells against hypoxia and serum deprivation-induced apoptosis via the PI3K/Akt and ERK1/2 pathways
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DOI：
10.1631/jzus.b1001007
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发表时间：
2010-08-01
影响因子：
5.1
通讯作者：
Wang, Jian-an
中科院分区：
文献类型：
--
作者：
Gao, Feng;Hu, Xin-yang;Wang, Jian-an
骨髓间充质干细胞（Mesenchymal stem cell，MSC）移植已显示出修复缺血和梗死心肌的治疗潜力，但其效果受到宿主心脏微环境中供体细胞凋亡和丢失的限制。本研究旨在探讨热休克蛋白90（Hsp 90）对缺氧缺血清诱导的大鼠骨髓间充质干细胞凋亡的保护作用及其可能机制。通过3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四唑（MTT）测定法测定细胞活力。Hoechst 33258核染色和Annexin V/PI染色流式细胞术分析细胞凋亡。实时荧光定量PCR检测Toll样受体4（TLR-4）和V-erb-b2型成红细胞白血病病毒癌基因同源物2（ErbB 2）基因表达。Western blot检测caspase-3、Bcl-2、Bcl-xL、Bax、total-ERK、phospho-ERK、total-Akt、phospho-Akt和Hsp 90蛋白水平。用分光光度法测定一氧化氮的产生。热休克蛋白90可提高骨髓间充质干细胞的活力，并保护骨髓间充质干细胞免受血清剥夺和缺氧诱导的凋亡。Hsp 90的保护作用不仅通过下调细胞膜TLR-4和ErbB 2受体，进而激活其下游的PI 3 K/Akt和ERK 1/2通路，提高Bcl-2/Bax和Bcl-xL/Bax表达，降低caspase-3的表达，而且还增强了MSCs的旁分泌作用。这些发现证明了细胞移植中针对MSC凋亡的新颖且有效的治疗策略。
Mesenchymal stem cell (MSC) transplantation has shown a therapeutic potential to repair the ischemic and infracted myocardium, but the effects are limited by the apoptosis and loss of donor cells in host cardiac microenvironment. The aim of this study is to explore the cytoprotection of heat shock protein 90 (Hsp90) against hypoxia and serum deprivation-induced apoptosis and the possible mechanisms in rat MSCs. Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis was assessed by Hoechst 33258 nuclear staining and flow cytometric analysis with annexin V/PI staining. The gene expression of Toll-like receptor-4 (TLR-4) and V-erb-b2 erythroblastic leukemia viral oncogene homolog 2 (ErbB2) was detected by real-time polymerase chain reaction (PCR). The protein levels of cleaved caspase-3, Bcl-2, Bcl-xL, Bax, total-ERK, phospho-ERK, total-Akt, phospho-Akt, and Hsp90 were detected by Western blot. The production of nitric oxide was measured by spectrophotometric assay. Hsp90 improves MSC viability and protects MSCs against apoptosis induced by serum deprivation and hypoxia. The protective role of Hsp90 not only elevates Bcl-2/Bax and Bcl-xL/Bax expression and attenuates cleaved caspase-3 expression via down-regulating membrane TLR-4 and ErbB2 receptors and then activating their downstream PI3K/Akt and ERK1/2 pathways, but also enhances the paracrine effect of MSCs. These findings demonstrated a novel and effective treatment strategy against MSC apoptosis in cell transplantation.