非洲传统草药Hypoxis Hemerocallidea提取物对糖尿病骨骼肌AMPK信号通路作用机制研究  

Study on Action Mechanism of African Traditional Herb Hypoxis Hemerocallidea Extracts on AMPK Signal Pathway of Skeletal Muscles in Diabetes

作　　者：郭璇[1] 孙文[2,3,4] 刘铜华[2,3,4,5] 吴丽丽[2,3,4] 许光远[1] 张岩[1] 穆晓红[2,3,6] 郭翔宇[1,2,4] 徐暾海[2,3,7] 秦灵灵[2,3,8] 

机构地区：[1]北京中医药大学东方医院,北京100029 [2]北京中医药大学教育部中医养生学重点实验室,北京100029 [3]北京中医药大学北京市中医养生学重点实验室,北京100029 [4]北京中医药大学中医养生学研究所,北京100029 [5]北京中医药大学研究生院,北京100029 [6]北京中医药大学东直门医院,北京100700 [7]北京中医药大学中药学院,北京100102 [8]北京中医药大学科技处,北京100029

出　　处：《世界科学技术-中医药现代化》2015年第6期1157-1163,共7页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：科学技术部国家国际科技合作与交流项目(2012DFG31550):南非传统草药抗糖尿病活性成分筛选及机制研究;负责人:刘铜华;北京中医药大学创新团队项目(2011-CXTD-19):中医药防治糖尿病研究团队;负责人:刘铜华

摘　　要：目的:探讨Hypoxis Hemerocallidea(AP)对糖尿病大鼠骨骼肌AMPK信号通路的影响。方法:1 40只雄性SD大鼠,10只作为正常组,30只高脂喂养1月,加一次性腹腔注射STZ造模,造模成功后分为模型组、盐酸吡格列酮组、AP组,分组灌胃5周。取骨骼肌组织包埋切片进行免疫组化;利用Western Blot检测骨骼肌p-AMPKα、p-AS160、GLUT4蛋白表达情况;2使用100 mmol·L-1葡萄糖诱导建立C2C12骨骼肌细胞胰岛素抵抗模型,设为模型组;另加AP含药血清设置处理组;对照组为正常细胞。检测各组葡萄糖消耗量,细胞增殖情况,检测SOD、MDA含量,采用Western Blot、RT-PCR检测各组p-AMPKα、p-AS160、GLUT4蛋白表达情况。结果:1与正常组比较,AP能上调DM大鼠骨骼肌组织p-AMPKα蛋白量(P<0.01),增加骨骼肌AS160的磷酸化水平(P<0.01),上调GLUT4表达(P<0.01);2与正常组比较,高糖造成了C2C12骨骼肌细胞活性下降,葡萄糖消耗量减低(P<0.05),SOD降低(P<0.01),MDA增加(P<0.01),p-AMPKα、p-AS160、GLUT4蛋白表达降低(P<0.01)。干预48 h后,AP含药血清组C2C12骨骼肌细胞SOD均显著增高(P<0.01),MDA含量降低(P<0.05),提高了AMPKα、AS160的磷酸化水平(P<0.01),增加GLUT4蛋白表达(P<0.01)。结论:诱导AMPKα、AS160磷酸化促进GLUT4表达可能是AP改善糖尿病骨骼肌胰岛素抵抗的作用机制之一。This study was aimed to investigate the action mechanism of Hypoxis Hemerocallidea（African Potato, AP） on the AMPK signal pathway of skeletal muscles in diabetic rats. Among 40 male SD rats, 10 rats were used as the normal group, and the other 30 rats were fed with high-fat food for one month, and then injected with STZ for the model establishment. After the successful model establishment, rats were divided into the model group, pioglitazone hydrochloride group and the AP group. Intragastric administration was given for 5 weeks in each group. Then, the skeletal muscle tissues were embedded and sliced for immunohistochemistry test. The protein expression of p-AMPKα, p-AS16 and GLUT4 in skeletal muscles was detected by western blot. The 100 mmol·L^-1 glucose was used in the establishment of C2C12 skeletal muscle cells insulin resistance model. AP drug-containing serum was used in the establishment of the treatment group. The control group was the normal cells. Glucose consumption, cell proliferation, SOD content, and MDA content were detected. And the protein expressions of p-AMPKα, p-AS160, GLUT4 were detected with the western blot and RT-PCR. The results showed that compared with the normal group, AP can up-regulate p-AMPKa protein express（P〈0.01）, increase skeletal AS160 phosphorylation level（P〈0.01）, and up-regulate the GLUT4 level（P〈0.01）. Compared with the normal group, the high glucose caused the decrease of C2C12 skeletal muscle cell activity and the decrease of glucose consumption（P〈0.05）, decrease of SOD, increase of MDA（P〈0.01）, and the decrease of p-AMPKα, p-AS160, GLUT4 protein expression（P〈0.01）. After 48 h intervention, the SOD of C2C12 skeletal muscle cells in the AP drug-containing serum group was significantly increased（P〈0.01）, the MDA content was decreased（P〈0.05）, the AMPKa and AS160 phosphorylation levels were increased（P〈0.01）, the GLUT protein expression was increased（P〈0.01）. It was concluded that the induced AMPKa and AS16

关 键 词：非洲马铃薯 糖尿病 骨骼肌 AMPK 实验研究 

分 类 号：R285.5[医药卫生—中药学]