基于AMPK-ACC信号通路探讨加味温胆汤及其拆方改善营养性肥胖大鼠瘦素抵抗的机制

According to WHO,at least 2,800,000 adults died of overweight or obese each year and that makes scientific prevention of obesity become the social issue of common concern to the people all over the world.Traditional Chinese medicine(TCM) believes that obesity is closely related to deficiency,phlegm and static blood.Western medical research shows that leptin resistance is one of the key factors leading to obesity,and AMPK-ACC signal pathway can decrease fat deposition as well as improve the body sensitivity to leptin by regulating fatty acid synthesis and oxidation.It is Duan Fujin,the Chinese medicine master,that developed modified Wendan Decoction specially for obesity by adding Radix Astragali seuHedysari,Semen Sinapis Albae and Radix Salviae Miltiorrhizae to the famous phlegm-eliminating formula Wendan Decoction.Previous studies showed this decoction can obviously reduce the amount of fat around the organs, improve the degree of fatty degeneration of hepatic cells, reduce the size of fat cells, and against the higher serum leptin level of rats induced by high fat diet significantly.That leads to our conclusion that improving obesity rat leptin resistance is one of the mechanism of modified Wendan Decoction to lose weight,and this mechanism may be related to the regulation of leptin though AMPK-ACC Signaling pathway.This project will clear.the targets by analysing the decoction’s impact on Leptin-AMPK-ACC-MA-CPTⅠpathway indexes of adult rat and young rats both including.the two sexes,which may provide experimental basis for the clinical application and the development of new drugs or health care products.

WHO称每年有280万成人死于超重或肥胖,科学的防治肥胖成为全世界人民共同关注的社会问题。中医学认为肥胖与虚、痰、瘀密切相关;西医学研究表明:瘦素抵抗是导致肥胖的关键因素之一,而AMPK-ACC信号通路可调控脂肪酸合成与氧化而减少脂肪沉积,提高机体对瘦素的敏感性。加味温胆汤是针对肥胖虚、痰、瘀病机，根据国医大师段富津的临床经验,在祛痰名方“温胆汤”基础上加入黄芪、白芥子、丹参演化而来,前期研究表明:该方可明显减少脏器周围脂肪量、改善肝细胞脂肪变性程度,缩小脂肪细胞体积,显著对抗高脂饮食所致大鼠血清瘦素水平的升高,故课题组认为:改善肥胖鼠瘦素抵抗是加味温胆汤减肥的机制之一,其机理可能与调控瘦素后AMPK-ACC通路有关,本课题拟通过分析其对雌雄性幼鼠、成年鼠Leptin-AMPK-ACC-MA-CPTⅠ通路指标的影响,明确其作用靶点,为临床应用该方防治肥胖或开发减肥新药、保健品奠定实验基础。

通过观察加味温胆汤对营养性肥胖成年大鼠、幼鼠AMPK-ACC通路指标的影响，揭示其抗营养性肥胖的机制，结果表明：1）对雄性成年大鼠的影响：与模型对照组比较，加味温胆汤高剂量组可显著降低大鼠Lee’s指数、BMI指数及血清GLU、LEP、INS含量；加味温胆汤中剂量组可升高大鼠血清ADP含量；加味温胆汤低剂量组可降低大鼠骨骼肌组织AMPKβ、ACC含量；各给药组均可明显减小大鼠脂肪细胞体积，减轻肝细胞脂肪变性程度。2）对雌性成年大鼠的影响：与模型对照组比较，加味温胆汤高剂量组可显著下调雌性大鼠腓肠肌组织CPT1及AMPKα2 mRNA的表达；中剂量组可显著下调腓肠肌组织CPT1，下丘脑组织LEP mRNA、MCA mRNA，脂肪组织AMPKα1 mRNA及腓肠肌组织AMPKα2 mRNA的表达；低剂量组可显著下调血清LEP，下丘脑组织LEP mRNA、MCA mRNA，脂肪组织AMPKα1 mRNA，腓肠肌组织AMPKα2 mRNA及肝组织AMPKβ、ACC2 mRNA的表达。3）对雄性幼鼠的影响：与模型对照组比较，加味温胆汤高剂量组可提高血清Glu、HDL-C及骨骼肌AMPK mRNA的表达水平，降低血清TG和骨骼肌ACC mRNA的表达水平及Lee’s指数；加味温胆汤中剂量组可升高血清Glu、LEP及骨骼肌LEP mRNA、AMPK mRNA、MLYCDmRNA、MCD的表达水平，降低血清LDL-C和骨骼肌ACC mRNA的表达水平；加味温胆汤低剂量组可显著提高血清Glu、LEP，降低骨骼肌ACC mRNA的表达水平。4）对雌性幼鼠的影响：与模型对照组相比，加味温胆汤高剂量组可下调血清TG、LDL-C及骨骼肌INS含量，上调ADP含量和骨骼肌COX的表达水平，减轻肝细胞脂肪变性；中剂量组可降低幼鼠脂肪指数和Lee’s指数，下调血清TC、TG、LEP，骨骼肌INS含量及骨骼肌IRS2 mRNA的表达，上调骨骼肌SLC2A4 mRNA并增强骨骼肌COX表达；低剂量组可降低幼鼠摄食高脂乳剂量，下调血清TG、TC、LDL-C及骨骼肌INS含量，减轻肝细胞脂肪变性，增强骨骼肌COX表达。课题为临床应用该方防治肥胖或开发减肥新药、保健品奠定实验基础。