代综方对C2C12骨骼肌细胞葡萄糖摄取及GLUT4蛋白表达的影响
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摘要
目的:研究代综方(dai zong fang,DZF)对骨骼肌细胞葡萄糖摄取及葡萄糖转运体4(glucose transporter 4,GLUT4)蛋白表达的影响。方法:采用含2%马血清(horse serum,HS)的DMEM培养液诱导C2C12骨骼肌细胞分化为肌管,荧光标记的葡萄糖类似物2-NBDG测定葡萄糖摄取,对2-NBDG的量效时效关系进行测定;采用不同剂量DZF干预分化的C2C12细胞,检测其对细胞基础状态及胰岛素刺激状态下葡萄糖摄取的影响;Western blot检测DZF对C2C12细胞GLUT4总蛋白表达及膜转位的影响。结果:在基础状态或胰岛素刺激下,DZF低、中、高剂量均可提高骨骼肌葡萄糖摄取,差异有统计学意义(P<0.05)。基础状态下,DZF能促进GLUT4总蛋白及膜转位蛋白的表达(P<0.05);胰岛素刺激状态下,DZF对GLUT4总蛋白表达无显著影响(P>0.05),但仍可促进GLUT4膜转位蛋白的表达(P<0.05)。结论:DZF可增加骨骼肌细胞葡萄糖摄取,具有类胰岛素作用,其作用机制可能与促进GLUT4蛋白向细胞膜转位有关。
Objectives:To investigate the effects of dai zong fang(DZF)on glucose uptake and glucose transporter 4 protein expression in skeletal muscle cells. Methods:C2 C12 skeletal muscle cells were differentiated into myotubes by incubating in DMEM with 2%horse serum;glucose uptake in C2 C12 myotubes were measured by 2-NBDG;the dose effect and time effect of 2-NBDG were analyzed. After the intervention of different concentrations of DZF,glucose uptake in C2 C12 myotubes were analyzed in basal and insulin-stimulated state. Effects of DZF on GLUT4 protein expression and translocation were analyzed by Western blot. Results:Both in basal and insulin-stimulated state,different concentrations of DZF could stimulate the glucose uptake in skeletal muscle cells,with statistically significant difference(P<0.05). In the basal state,DZF significantly increased total and membrane GLUT4 protein expression(P<0.05). In the insulin-stimulated state,DZF had no significant effect on the total GLUT4 protein expression(P>0.05),however,it could significantly promote the membrane GLUT4 protein expression(P<0.05). Conclusion:DZF could increase glucose uptake in skeletal muscle cells and have insulin-like function,the mechanisms might be related to the promotion of GLUT4 translocation.
Objectives:To investigate the effects of dai zong fang(DZF)on glucose uptake and glucose transporter 4 protein expression in skeletal muscle cells. Methods:C2 C12 skeletal muscle cells were differentiated into myotubes by incubating in DMEM with 2%horse serum;glucose uptake in C2 C12 myotubes were measured by 2-NBDG;the dose effect and time effect of 2-NBDG were analyzed. After the intervention of different concentrations of DZF,glucose uptake in C2 C12 myotubes were analyzed in basal and insulin-stimulated state. Effects of DZF on GLUT4 protein expression and translocation were analyzed by Western blot. Results:Both in basal and insulin-stimulated state,different concentrations of DZF could stimulate the glucose uptake in skeletal muscle cells,with statistically significant difference(P<0.05). In the basal state,DZF significantly increased total and membrane GLUT4 protein expression(P<0.05). In the insulin-stimulated state,DZF had no significant effect on the total GLUT4 protein expression(P>0.05),however,it could significantly promote the membrane GLUT4 protein expression(P<0.05). Conclusion:DZF could increase glucose uptake in skeletal muscle cells and have insulin-like function,the mechanisms might be related to the promotion of GLUT4 translocation.
引文
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[4]赵玉雪,朱晓云,曹松华,等.3 398例代谢综合征患者证候学分析[J].辽宁中医杂志,2016,59(1):5-7.
[5]朱晓云,王朋倩,刘喜明.代综方对3T3-L1脂肪细胞糖脂代谢的作用研究[J].世界中西医结合杂志,2015,10(2):175-178.
[6]朱晓云,楚小燕,王兆礼,等.代综方对3T3-L1脂肪细胞分化及相关基因表达的影响[J].重庆医科大学学报,2015,37(7):1016-1020.
[7]董菲菲,朱晓云,张俊龙,等.代综方对油酸诱导的Hep G2细胞内脂质堆积的影响[J].世界中西医结合杂志,2015,10(3):330-334.
[8]叶林,陆凯,张冬颖,等.2-NBDG检测H9c2心肌细胞葡萄糖摄取能力的研究[J].第三军医大学学报,2012,34(15):1533-1537.
[9]Millon SR,Ostrander JH,Brown JQ,et al.Uptake of 2-NBDG as a method to monitor therapy response in breast cancer cell lines[J].Breast Cancer Research and Treatment,2010,126(1):55-62.
[10]胡慧,单秀红,朱伟,等.2-NBDG用于乳腺癌MDA-MB-231细胞荧光检测研究[J].江苏大学学报:医学版,2010,20(3):216-219.
[11]Baron AD,Brechtel G,Wallace P,et al.Rates and tissue sites of non-insulin-and insulin-mediated glucose uptake in humans[J].Am J Physiol,1988,255(6P1):E769-E774.
[12]Li HB,Yang YR,Mo ZJ,et al.Silibinin improves palmitate-induced insulin resistance in C2C12 myotubes by attenuating IRS-1/PI3K/Akt pathway inhibition[J].Brazilian Journal of Medical and Biological Research,2015,48(5):440-446.
[13]Maurya CK,Singh R,Jaiswal N,et al.4-Hydroxyisoleucine ameliorates fatty acid-induced insulin resistance and inflammatory response in skeletal muscle cells[J].Molecular and Cellular Endocrinology,2014,395(1-2):51-60.