世界科学技术-中医药现代化
世界科學技術-中醫藥現代化
세계과학기술-중의약현대화
WORLD SCIENCE AND TECHNOLOGY-MODERNIZATION OF TRADITIONAL CHINESE MEDICINE
2015年
6期
1157-1163
,共7页
郭璇%孙文%刘铜华%吴丽丽%许光远%张岩%穆晓红%郭翔宇%徐暾海%秦灵灵
郭璇%孫文%劉銅華%吳麗麗%許光遠%張巖%穆曉紅%郭翔宇%徐暾海%秦靈靈
곽선%손문%류동화%오려려%허광원%장암%목효홍%곽상우%서돈해%진령령
非洲马铃薯%糖尿病%骨骼肌%AMPK%实验研究
非洲馬鈴藷%糖尿病%骨骼肌%AMPK%實驗研究
비주마령서%당뇨병%골격기%AMPK%실험연구
Hypoxis Hemerocallidea%diabetes%skeletal muscle%AMPK%experimental research
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:①40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。
目的:探討Hypoxis Hemerocallidea(AP)對糖尿病大鼠骨骼肌AMPK信號通路的影響。方法:①40隻雄性SD大鼠,10隻作為正常組,30隻高脂餵養1月,加一次性腹腔註射STZ造模,造模成功後分為模型組、鹽痠吡格列酮組、AP組,分組灌胃5週。取骨骼肌組織包埋切片進行免疫組化;利用Western Blot檢測骨骼肌p-AMPKα、p-AS160、GLUT4蛋白錶達情況;②使用100 mmol·L-1葡萄糖誘導建立C2C12骨骼肌細胞胰島素牴抗模型,設為模型組;另加AP含藥血清設置處理組;對照組為正常細胞。檢測各組葡萄糖消耗量,細胞增殖情況,檢測SOD、MDA含量,採用Western Blot、RT-PCR檢測各組p-AMPKα、p-AS160、GLUT4蛋白錶達情況。結果:①與正常組比較,AP能上調DM大鼠骨骼肌組織p-AMPKα蛋白量(P<0.01),增加骨骼肌AS160的燐痠化水平(P<0.01),上調GLUT4錶達(P<0.01);②與正常組比較,高糖造成瞭C2C12骨骼肌細胞活性下降,葡萄糖消耗量減低(P<0.05),SOD降低(P<0.01),MDA增加(P<0.01),p-AMPKα、p-AS160、GLUT4蛋白錶達降低(P<0.01)。榦預48 h後,AP含藥血清組C2C12骨骼肌細胞SOD均顯著增高(P<0.01),MDA含量降低(P<0.05),提高瞭AMPKα、AS160的燐痠化水平(P<0.01),增加GLUT4蛋白錶達(P<0.01)。結論:誘導AMPKα、AS160燐痠化促進GLUT4錶達可能是AP改善糖尿病骨骼肌胰島素牴抗的作用機製之一。
목적:탐토Hypoxis Hemerocallidea(AP)대당뇨병대서골격기AMPK신호통로적영향。방법:①40지웅성SD대서,10지작위정상조,30지고지위양1월,가일차성복강주사STZ조모,조모성공후분위모형조、염산필격렬동조、AP조,분조관위5주。취골격기조직포매절편진행면역조화;이용Western Blot검측골격기p-AMPKα、p-AS160、GLUT4단백표체정황;②사용100 mmol·L-1포도당유도건립C2C12골격기세포이도소저항모형,설위모형조;령가AP함약혈청설치처리조;대조조위정상세포。검측각조포도당소모량,세포증식정황,검측SOD、MDA함량,채용Western Blot、RT-PCR검측각조p-AMPKα、p-AS160、GLUT4단백표체정황。결과:①여정상조비교,AP능상조DM대서골격기조직p-AMPKα단백량(P<0.01),증가골격기AS160적린산화수평(P<0.01),상조GLUT4표체(P<0.01);②여정상조비교,고당조성료C2C12골격기세포활성하강,포도당소모량감저(P<0.05),SOD강저(P<0.01),MDA증가(P<0.01),p-AMPKα、p-AS160、GLUT4단백표체강저(P<0.01)。간예48 h후,AP함약혈청조C2C12골격기세포SOD균현저증고(P<0.01),MDA함량강저(P<0.05),제고료AMPKα、AS160적린산화수평(P<0.01),증가GLUT4단백표체(P<0.01)。결론:유도AMPKα、AS160린산화촉진GLUT4표체가능시AP개선당뇨병골격기이도소저항적작용궤제지일。
This study was aimed to investigate the action mechanism ofHypoxis 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 AS160 phosphorylation promoted GLUT 4 expression may be one of the action mechanism of insulin resistance of skeletal muscles in diabetes.