Effect of Huatan Qushi Huoxue prescription on the ADPN/AMPK/ACC signaling pathway in rats with nonalcoholic steatohepatitis
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摘要:
目的研究化痰祛湿活血方对非酒精性脂肪性肝炎(NASH)大鼠脂联素(ADPN)/腺苷酸活化蛋白激酶(AMPK)/乙酰辅酶A羧化酶(ACC)通路的影响。方法 72只SD大鼠随机分为空白组、模型组、对照组及高、中、低剂量组,除空白组外,其余5组以高脂饲料联合四环素腹腔注射建立NASH模型,造模第2周分别给予化痰祛湿活血方高、中、低剂量和多烯磷脂酰胆碱(易善复)胶囊对照治疗。实验结束时,观察肝匀浆游离脂肪酸(FFA)含量,ELISA及RT-q PCR法测肝组织ADPN、脂联素受体2(AdipoR2)、AMPK、ACC、肉毒碱棕榈酰转移酶1(CPT-1)的基因与蛋白表达。多组间数据比较采用方差分析,方差齐者进一步两组间比较采用LSD-t检验,方差不齐者采用Dunnett’s T3法。结果与对照组比较,高剂量组能明显降低肝匀浆FFA[(229.07±46.62)μmol/gprot vs(382.75±30.93)μmol/gprot]、肝组织ACC的基因(1.66±0.04 vs 2.40±0.37)、蛋白[(512.20±60.70)pg/g vs(756.72±61.21)pg/g]表达(P...
Abstract:Objective To investigate the effect of Huatan Qushi Huoxue prescription on the adiponectin (ADPN) /adenosine monophosphate-activated protein kinase (AMPK) /acetyl-Co A carboxylase (ACC) signaling pathway in rats with nonalcoholic steatohepatitis (NASH) .Methods A total of 72 male Sprague-Dawley rats were randomly divided into blank group, model group, control group, and high-, middle-, and low-dose Huatan Qushi Huoxue prescription groups.All rats except those in the blank group were given high-fat diet combined with intraperitoneal injection of tetracycline to establish a rat model of NASH.At week 2 after model establishment, the Huatan Qushi Huoxue prescription groups were treated with high-, middle-, and low-dose Huatan Qushi Huoxue prescription, and polyene phosphatidylcholine (Yishanfu) capsules were given as control.At the end of the experiment, the content of free fatty acid (FFA) in liver homogenate was measured, and ELISA and RT-q PCR were used to measure the protein and mRNA expression of ADPN, adiponectin receptor-2 (AdipoR2) , AMPK, ACC, and carnitine palmitoyltransferase-1 (CPT-1) in liver tissue.An analysis of variance was used for comparison of data between multiple groups;the least significant difference t-test was used for comparison of data with homogeneity of variance between two groups, and the Dunnett's T3 method was used for comparison of data with heterogeneity of variance.Results Compared with the control group, the high-dose group had significant reductions in FFA level in liver homogenate (229.07±46.62 μmol/gprot vs 382.75 ±30.93 μmol/gprot, P<0.01) and mRNA and protein expression of ACC in liver tissue (mRNA:1.66±0.04 vs 2.40±0.37, P<0.01;protein:512.20±60.70 pg/g vs 756.72±61.21 pg/g, P<0.01) , as well as significant increases in the mRNA and protein expression of ADPN (mRNA:1.53±0.31 vs 0.75±0.11, P<0.01;protein:22.23±2.75 μg/g vs 14.01±2.17 μg/g, P<0.01) , AdipoR2 (mRNA:1.38±0.33 vs 0.61±0.09, P<0.01;protein:128.41±10.46 ng/g vs 94.62±6.88 ng/g, P<0.01) , AMPK (mRNA:1.43±0.39 vs 0.78±0.04, P<0.01;protein:1353.79±59.52 μg/g vs 1107.19±56.78 μg/g, P<0.01) , and CPT-1 (mRNA:1.39±0.31 vs 0.52±0.04, P<0.01;protein:175.54±9.91 U/g vs 128.74±12.74 U/g, P<0.01) .Conclusion Huatan Qushi Huoxue prescription can effectively activate the ADPN/AMPK/ACC signaling pathway, which may affect the β oxidation and synthesis of fatty acids and reduce fat deposition in hepatocytes.
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