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目的:研究ADIPONECTIN对C2C12肌细胞糖原合成和葡萄糖氧化的影响。方法:用阳离子脂质体介导转染和随后G418筛选建守稳定转染小鼠ADIPONECTIN CDNA真核表达质粒(PCDNA3．0-MAD)及空载PCDNA3．0的C2C12 细胞株并鉴定。C2C12肌细胞糖代谢实验分对照组、空载体组和PCDNA3．0-MAD(MAD)组共3组进行,每组又分 0、0．5、5、100 NMOL／L胰岛素刺激4个亚组。通过液闪测定细胞合成的糖原中[14C]的放射活性和氧化产生的[14CO2], 分别检测肌细胞的糖原合成和葡萄糖氧化情况。结果:WESTERN BLOTTING和免疫组化检测证实MAD组细胞表达 ADIPONECTIN蛋白。MAD组葡萄糖氧化量随胰岛素浓度增加的速率较其它两组快,对照、空载体和MAD组线性回归系数分别为23．34、23．23和26．06。MAD组C2C12肌细胞基础状态下和胰岛素刺激下的葡萄糖氧化和糖原合成与其它两组无显著差异(P>0．05)。结论:转染ADIPONECTIN基因对C2C12肌细胞葡萄糖氧化和糖原合成无显著影响。
AIM: To investigate the effects of ADIPONECTIN on glycogen synthesis and glucose oxidation in C2C12 myocytes. METHODS: The cationic liposome-mediated transfection and subsequent G418 screening of stable and stable expression of mouse ADIPONECTIN CDNA eukaryotic expression plasmid (PCDNA3.0-MAD) and empty PCDNA3.0 C2C12 cell line and identified. C2C12 muscle cell glucose metabolism test points control group, empty vector group and PCDNA3.0-MAD (MAD) group of three groups, each group is divided into 0,0.5,5,100 NMOL / L insulin stimulation 4 subgroups . The radioactivity of [14C] and the [14CO2] produced by oxidation were measured by liquid scintillation to detect the glycogen synthesis and glucose oxidation of muscle cells, respectively. Results: Western blotting and immunohistochemistry confirmed that MAD cells expressed ADIPONECTIN protein. The rate of glucose oxidation increased with insulin concentration in MAD group compared with the other two groups. The linear regression coefficients of control, empty vector and MAD group were 23.34,23.23 and 26.06 respectively. Glucose oxidation and glycogen synthesis in C2C12 myocytes of MAD group were not significantly different from those in other two groups (P> 0.05). CONCLUSION: Transfection of ADIPONECTIN gene has no significant effect on glucose oxidation and glycogen synthesis in C2C12 myocytes.