活性维生素D对白介素-6作用下成肌细胞增殖分化的影响及其机制研究.docVIP

活性维生素D3对白介素-6作用下肌原细胞增殖和分化的影响及机制研究 王会玲 李燕 徐凯 刘楠梅 田军 张金元 上海解放军第四五五医院肾脏科 南京军区肾脏病研究所 上海 200052 基金项目：上海浦江人才基金（10PJD016） 作者单位： 上海解放军第四五五医院肾脏科 南京军区肾脏病研究所 通信作者：王会玲，Email. viollla@163.com 目的 探讨1,25(OH)活性vitD3（1,25D3）对白介素－6（interleukin-6, IL-6）刺激下成肌细胞增殖和分化的影响及机制。方法 体外培养小鼠C2C12肌细胞株，2%马血清使细胞分化为肌索，给予IL-6~100ng/ml)和1,25-D3刺激细胞，光镜下观察细胞和肌管形态；Western印迹法检测vitD3 受体（VDR）、成肌分子MyoD、 Myogenin和肌动蛋白重链（MHC）等蛋白表达水平，荧光定量PCR检测myostatin、Atrogin-1MyoD、Myogenin mRNA转录水平。生肌调节因子慢性肾脏病白介素－6 1,25(OH)2-vitamin D3 prevents interleukin-6 induced muscle atrophy by stimulating differentiation and inhibiting myostatin expression in C2C12 muscle cells. Wang Huiling, Li Yan, Xu Kai, Liu Nanmei, TianJjun, Zhang Jinyuan Department of Nephrology, 455th Hospital of PLA, The Institute of Nephrology of Nanjing Military Command, Shanghai 200052, China This work was supported by the Project of Shanghai Pujiang Program (No.10PJD016） Corresponding author: WANG Hui-ling, Email: viollla@163.com Objective To investigate the effect and mechanism of 1,25-dihydroxy-vitamin D (1,25-D3) on proliferation and differentiation of C2C12 myoblast cells exposure with interleukin-6(IL-6). Methods Cultured mouse C2C12 cells,2% horse serum induced cells differentiated into myotubes. After treated with IL-6( 0~100ng/ml) or 1,25-D3(100nM), the cells proliferation and myotube’s morphology was observed by light microscopy, the protein expression of vitamin D receptor(VDR) and myogenic regulatory factors such as MyoD and Myogenin and actin heavy chain (MHC) were measured by Western blotting. The mRNA transcription levels of muscle atrophy molecular such as myostatin and Atrogin-1, the mRNA of MyoD and Myogenin were quantitative by real time-PCR. Results The growth myoblasts were like fusiform, proliferation to 80% dish after 72h. After changed medium with 2% horse serum, the cells differentiate into myotube. When chronic exposure to IL-6(20ng/ml) , the cells proliferated slowly, and more necrotic cells was observed when