Department of Rehabilitation Medicine, Linyi Central Hospital, No.17, Jiankang Road, Linyi, 276400, Shandong, China.
Department of Gynaecology, Linyi Central Hospital, Linyi, 276400, Shandong, China.
J Mol Neurosci. 2019 Mar;67(3):400-410. doi: 10.1007/s12031-018-1243-7. Epub 2019 Jan 15.
We attempted to explore the possible effects of SOCS3 (suppressor of cytokine signaling 3)-modified bone marrow mesenchymal stem cells (BMSCs) on the hypoxic injury of rat adrenal gland pheochromocytoma (PC-12) cells. PC12 cells were cultured with EGFP (enhanced green fluorescent protein)-BMSCs and SOCS3-BMSCs respectively under hypoxia in vitro and classified into control, hypoxia, EGFP-BMSCs, and SOCS3-BMSC groups. CCK-8, Hoechst 33258 staining, and Annexin V-FITC/PI staining were assessed to measure the viability and apoptosis of hypoxia-induced PC12 cells. The JAK/STAT3 pathway- and apoptosis-related proteins were identified by Western blot. Finally, rat models of permanent middle cerebral artery occlusion (pMCAO) were established to verify the potential influences of SOCS3-BMSCs in vivo. SOCS3-modified BMSCs can stably express SOCS3 protein. EGFP-BMSCs, especially SOCS3-BMSCs, can improve cell viability and SOD content, and reduce cell apoptosis, LDH viability, and MDA content in hypoxia-induced PC12 cells (all P < 0.05). Besides, EGFP-BMSCs and SOCS3-BMSCs decreased cleaved caspase-3 level and increased Bcl-2/Bax of hypoxia-induced PC12 cells, while SOCS3-BMSCs could also elevate SOCS3 protein and reduce p-STAT3 protein level in hypoxia-induced PC12 cells (all P < 0.05). In vivo experiments confirmed that EGFP-BMSCs, particularly SOCS3-BMSCs, could ameliorate infarct size and inhibit neuronal apoptosis of different degrees in pMACO rats (all P < 0.05). SOCS3-modified BMSCs can alleviate oxidative stress, improve cell viability, and reduce neuronal apoptosis by downregulation of JAK/STAT3 pathway, thereby exerting the neuroprotective role in ischemic brain injury.
我们试图探索 SOCS3（细胞因子信号转导抑制因子 3）修饰的骨髓间充质干细胞（BMSCs）对大鼠肾上腺嗜铬细胞瘤（PC-12）细胞缺氧损伤的可能影响。将 PC12 细胞分别与 EGFP（增强型绿色荧光蛋白）-BMSCs 和 SOCS3-BMSCs 在体外缺氧条件下培养，并分为对照组、缺氧组、EGFP-BMSCs 组和 SOCS3-BMSC 组。通过 CCK-8、Hoechst 33258 染色和 Annexin V-FITC/PI 染色评估缺氧诱导的 PC12 细胞活力和凋亡。通过 Western blot 鉴定 JAK/STAT3 通路和凋亡相关蛋白。最后，建立大鼠永久性大脑中动脉闭塞（pMCAO）模型，以验证 SOCS3-BMSCs 在体内的潜在影响。SOCS3 修饰的 BMSCs 可以稳定表达 SOCS3 蛋白。EGFP-BMSCs，尤其是 SOCS3-BMSCs，可提高细胞活力和 SOD 含量，降低缺氧诱导的 PC12 细胞凋亡、LDH 活力和 MDA 含量（均 P < 0.05）。此外，EGFP-BMSCs 和 SOCS3-BMSCs 降低了缺氧诱导的 PC12 细胞中 cleaved caspase-3 的水平，增加了 Bcl-2/Bax 的水平，而 SOCS3-BMSCs 还可以提高缺氧诱导的 PC12 细胞中 SOCS3 蛋白的水平，降低 p-STAT3 蛋白的水平（均 P < 0.05）。体内实验证实，EGFP-BMSCs，特别是 SOCS3-BMSCs，可改善 pMACO 大鼠不同程度的梗死面积并抑制神经元凋亡（均 P < 0.05）。SOCS3 修饰的 BMSCs 通过下调 JAK/STAT3 通路减轻氧化应激，提高细胞活力，减少神经元凋亡，从而发挥对缺血性脑损伤的神经保护作用。
