基于PI3K/Akt信号通路探讨黄芪葛根配伍对糖尿病大鼠骨骼肌糖代谢的影响
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摘要
目的:基于PI3K/Akt信号通路中胰岛素(Ins)、胰岛素受体(InsR)、胰岛素受体底物-2(IRS-2)、磷脂酰肌醇-3激酶(PI3K)、蛋白激酶B(Akt2)、糖原合成酶激酶3β(GSK-3β)以及葡萄糖转运体-4(GLUT-4)的变化,探讨黄芪葛根配伍对糖尿病大鼠骨骼肌糖代谢的影响。方法:66只大鼠随机分为6组,除正常组外,其余各组均腹腔注射链脲佐菌素溶液诱导为糖尿病模型。检测大鼠随机血糖,并应用酶联免疫吸附测定(ELISA)法检测骨骼肌Ins、InsR及GLUT-4水平,以及应用实时荧光定量PCR技术检测骨骼肌IRS-2、PI3K、Akt2、GSK-3β、GLUT-4mRNA的表达。结果:黄芪组、葛根组及黄芪葛根配伍组随机血糖均降低,且配伍组血糖下降最明显。黄芪葛根配伍组InsR、GLUT-4、IRS-2mRNA、PI3KmRNA、Akt2mRNA、GLUT-4mRNA表达升高,GSK-3βmRNA表达减低。在调节骨骼肌Ins、InsR及PI3KmRNA方面,黄芪与葛根有交互作用。结论:黄芪、葛根均具有降低血糖之效,其降糖机制可能与Ins、InsR、IRS-2、PI3K、Akt2、GSK-3β、GLUT-4的表达变化相关。
Objective:To explore the effects of Astragali Radix-Puerariae Lobatae Radix compatibility on glycometabolism in skeletal muscle of diabetic rats,by means of changes in expressions of insulin( Ins),insulin receptor( InsR),insulin receptor substrate-2( IRS-2),phosphotidylinositol 3-kinase( PI3K),protein kinase B-2( Akt2),glycogen synthase kinase-3β( GSK-3β) and glucose transporter-4( GLUT-4) on PI3K/Akt pathway. Methods:Totally 66 rats were randomly divided into 6 groups.Except the normal group,the diabetics model was induced by intraperitoneal injection of streptozotocin in other groups. The level of blood glucose was detected. And the contents of Ins,InsR and GLUT-4 in rats were detected by ELISA. Real-time PCR was used to detect expressions of IRS-2 mRNA,PI3KmRNA,Akt2 mRNA,GSK-3βmRNA and GLUT-4 mRNA. Results:Astragali radix,puerariae lobatae radix and their compatibility groups lowered the level of blood glucose,especially those in the compatibility group. The compatibility increased the levels of InsR,GLUT-4,IRS-2 mRNA,PI3KmRNA,Akt2 mRNA and GLUT-4 mRNA and lowered GSK-3βmRNA expression. In regulating Ins,InsR and PI3KmRNA in rats skeletal muscle,Astragali Radix and puerariae Lobatae Radix had the interaction effect. Conclusion:Astragali Radix and puerariae Lobatae Radix decreased the level of blood glucose. The mechanism may be associated with regulatory effect of levels of Ins,InsR,IRS-2,PI3K,Akt2,GSK-3β and GLUT-4.
Objective:To explore the effects of Astragali Radix-Puerariae Lobatae Radix compatibility on glycometabolism in skeletal muscle of diabetic rats,by means of changes in expressions of insulin( Ins),insulin receptor( InsR),insulin receptor substrate-2( IRS-2),phosphotidylinositol 3-kinase( PI3K),protein kinase B-2( Akt2),glycogen synthase kinase-3β( GSK-3β) and glucose transporter-4( GLUT-4) on PI3K/Akt pathway. Methods:Totally 66 rats were randomly divided into 6 groups.Except the normal group,the diabetics model was induced by intraperitoneal injection of streptozotocin in other groups. The level of blood glucose was detected. And the contents of Ins,InsR and GLUT-4 in rats were detected by ELISA. Real-time PCR was used to detect expressions of IRS-2 mRNA,PI3KmRNA,Akt2 mRNA,GSK-3βmRNA and GLUT-4 mRNA. Results:Astragali radix,puerariae lobatae radix and their compatibility groups lowered the level of blood glucose,especially those in the compatibility group. The compatibility increased the levels of InsR,GLUT-4,IRS-2 mRNA,PI3KmRNA,Akt2 mRNA and GLUT-4 mRNA and lowered GSK-3βmRNA expression. In regulating Ins,InsR and PI3KmRNA in rats skeletal muscle,Astragali Radix and puerariae Lobatae Radix had the interaction effect. Conclusion:Astragali Radix and puerariae Lobatae Radix decreased the level of blood glucose. The mechanism may be associated with regulatory effect of levels of Ins,InsR,IRS-2,PI3K,Akt2,GSK-3β and GLUT-4.
引文
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[15]吕娟,陈莉娜,马欣.PI3K/Akt信号通路与胰岛素抵抗相关疾病的关系[J].国外医学(医学地理分册),2012,33(2):127-131.
[16]迟毓婧,李晶,管又飞,等.PI3K-Akt信号传导通路对糖代谢的调控作用[J].中国生物化学与分子生物学报,2010,26(10):879-885.
[17]李艳敏,刘烨,郝明芬,等.黄芪葛根汤总黄酮、总皂苷对糖尿病大鼠肝脏IL-12、IL-15的影响及其交互作用[J].世界科学技术-中医药现代化,2014,16(11):2475-2479.
[18]王春怡,李卫民,高英,等.黄芪葛根汤对2型糖尿病大鼠胰岛素抵抗及PPAR-γm RNA表达的影响[J].中国实验方剂学杂志,2012,18(5):152-155.
[19]王春怡,陈艳芬,李卫民,等.黄芪葛根汤对糖尿病及胰岛素抵抗小鼠血糖的影响[J].时珍国医国药,2011,22(11):2731-2732.
[2]曹琼丹,杨育红,于胜男,等.黄芪甲苷对Ⅰ型糖尿病大鼠心肌细胞PGC-1α和NRF-1表达的影响[J].中国药理学通报,2015,31(8):1096-1101.
[3]ChenW,Xia Y,Zhao X,et al.The critical role of Astraqalus polysaccharides for the improvement of PPARα[correction of PPARα]-mediated lipotoxicity in diabetic cardiomyopathy[J].Plos One,2012,7(10):e45541.
[4]Qi W,Niu J,Qin Q,et al.AstragalosideⅣattenuates glycated albumininduced epithelial-to-mesenchymal transition by inhibiting oxidative stress in renal proximal tubular cells[J].Cell Stress Chaperones,2014,19(18):105-114.
[5]张婧,刘长山,刘阳,等.黄芪对2型糖尿病大鼠胰岛素抵抗及肾脏损伤的影响[J].临床合理用药杂志,2013,6(10):54-55.
[6]崔蕾,李姝玉,柴新楼,等.黄芪和葛根素对2型糖尿病KKAy小鼠肝损害干预的实验研究[J].北京中医药大学学报,2012,35(3):189-193,218.
[7]郑曙琴,梁茂新.明代以前消渴病证治规律研究[J].中华中医药杂志,2009,24(12):1627-1629.
[8]陈秀芳,雷康福,董敏,等.葛根素对糖尿病大鼠心肌损伤的影响[J].中国病理生理杂志,2010,26(4):650-655.
[9]SEONG S H,ROY A,JUNG H A,et al.Protein tyrosine phosphatase1B and a-glucosidase inhibitory activities of Pueraria lobata root and its constituents[J].J Ethnopharmacol,2016,194:706-716.
[10]WU K,LIANG T,DUAN X,et al.Anti-diabetic effects of puerarin,isolated from Pueraria lobata(Willd.),on streptozotocin-diabetogenic mice through promoting insulin expression and ameliorating metabolic function[J].Food ChemToxicol,2013,60(10):341-347.
[11]师林,杨玉彬,黄颖娟,等.健脾化湿方对脾虚痰湿型肥胖2型糖尿病胰岛素抵抗大鼠骨骼肌PI3K/Akt通路的影响[J].中华中医药杂志,2018,33(2):530-534.
[12]Winnay J N,Dirice E,Liew C W,et al.p85αDeficiency protectsβ-cells from endoplasmic reticulum stress-induced apoptosis[J].Proc Natl Acad Sci USA,2014,111(3):1192-1197.
[13]Nelson V L,Jiang Y P,Dickman K G,et al.Adipose tissue insulin resistance due to loss of PI3K p110αleads to decreased energy expenditure and obesity[J].Am J Physiol Endocrinol Metab,2014,306(10):E1205-E1216.
[14]Hu X,Wang S,Xu J,et al.Triterpenoid saponins from Stauntonia chinensis ameliorate insulin resistance via the AMP-activated protein kinase and IR/IRS-1/PI3K/Akt pathways ininsulin-resistant HepG2 cells[J].Int J Mol sci,2014,15(6):10446-10458.
[15]吕娟,陈莉娜,马欣.PI3K/Akt信号通路与胰岛素抵抗相关疾病的关系[J].国外医学(医学地理分册),2012,33(2):127-131.
[16]迟毓婧,李晶,管又飞,等.PI3K-Akt信号传导通路对糖代谢的调控作用[J].中国生物化学与分子生物学报,2010,26(10):879-885.
[17]李艳敏,刘烨,郝明芬,等.黄芪葛根汤总黄酮、总皂苷对糖尿病大鼠肝脏IL-12、IL-15的影响及其交互作用[J].世界科学技术-中医药现代化,2014,16(11):2475-2479.
[18]王春怡,李卫民,高英,等.黄芪葛根汤对2型糖尿病大鼠胰岛素抵抗及PPAR-γm RNA表达的影响[J].中国实验方剂学杂志,2012,18(5):152-155.
[19]王春怡,陈艳芬,李卫民,等.黄芪葛根汤对糖尿病及胰岛素抵抗小鼠血糖的影响[J].时珍国医国药,2011,22(11):2731-2732.