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小檗碱对氧化应激诱导的大鼠骨髓间充质干细胞凋亡的保护作用。

Protective effect of berberine against oxidative stress-induced apoptosis in rat bone marrow-derived mesenchymal stem cells.

作者信息

Li Wangyang, Liu Yamei, Wang Bin, Luo Yiwen, Hu Nianhong, Chen Dongfeng, Zhang Xunchao, Xiong Yunpu

机构信息

Department of Traumatology, The Third Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine, Guangzhou, Guangdong 510240, P.R. China.

Department of Diagnostics of Traditional Chinese Medicine, Guangzhou University of Traditional Chinese Medicine, Guangzhou, Guangdong 510006, P.R. China.

出版信息

Exp Ther Med. 2016 Dec;12(6):4041-4048. doi: 10.3892/etm.2016.3866. Epub 2016 Nov 3.

Abstract

Bone marrow-derived mesenchymal stem cells (BMSCs) have the potential to be used for the treatment of delayed union, nonunion or persistent bone defects in MSC-based cell therapy. However, implantation of BMSCs into the fracture site is confronted with apoptosis on account of harsh conditions and oxidative stress. In the present study, the anti-apoptotic effects of berberine (BBR) on BMSCs subjected to hydrogen peroxide (HO) are investigated, and the potential underlying mechanisms are explored. Oxidative injury was induced by exposure to HO, and cell viability was assessed using a cell counting kit-8 assay. The apoptosis of BMSCs was measured by Hoechst 33258 and Annexin V-fluorescein isothiocyanate/propidium iodide assay. Reactive oxygen species staining and superoxide dismutase (SOD) assay were applied to assess the anti-oxidative effect of BBR. Finally, western blot was performed to measure the expression levels of phosphorylated (p)-Akt, B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax) and cleaved caspase-3. In the present study, it was identified that BBR remarkably attenuated HO-induced apoptotic cell death via quenching ROS production and increasing SOD activity. Further studies indicated that BBR can reduce apoptosis by upregulating the expression level of p-Akt and Bcl-2, and downregulating the expression levels of Bax and cleaved caspase-3. Taken together, the results of the present study demonstrate that pretreatment with BBR could alleviate HO-induced apoptosis in rat BMSCs .

摘要

骨髓间充质干细胞(BMSCs)在基于间充质干细胞的细胞治疗中具有用于治疗延迟愈合、骨不连或持续性骨缺损的潜力。然而,由于恶劣条件和氧化应激,将BMSCs植入骨折部位会面临细胞凋亡。在本研究中,研究了小檗碱(BBR)对过氧化氢(H₂O₂)处理的BMSCs的抗凋亡作用,并探讨了潜在的作用机制。通过暴露于H₂O₂诱导氧化损伤,并使用细胞计数试剂盒-8法评估细胞活力。通过Hoechst 33258和膜联蛋白V-异硫氰酸荧光素/碘化丙啶法检测BMSCs的凋亡情况。应用活性氧染色和超氧化物歧化酶(SOD)测定法评估BBR的抗氧化作用。最后,进行蛋白质免疫印迹法检测磷酸化(p)-Akt、B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)和裂解的半胱天冬酶-3的表达水平。在本研究中,发现BBR通过淬灭活性氧生成和增加SOD活性,显著减轻了H₂O₂诱导的凋亡性细胞死亡。进一步研究表明,BBR可通过上调p-Akt和Bcl-2的表达水平,下调Bax和裂解的半胱天冬酶-3的表达水平来减少细胞凋亡。综上所述,本研究结果表明,BBR预处理可减轻H₂O₂诱导的大鼠BMSCs凋亡。

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