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目的探讨化痰祛湿活血方对非酒精性脂肪性肝炎(non-alcoholic steatohepatitis,NASH)大鼠ADPN/AKT/NF-κB信号通路中关键蛋白ADPN、AdipoR2、p-AKT(ser473)、p-NF-κBp65(ser536)表达的影响及抗炎作用。方法 72只雄性SD大鼠,随机分为空白组、模型组、对照组及化痰祛湿活血方高剂量组、中剂量组、低剂量组,每组12只;空白组予普通饲料、不给药,其余5组予高脂饲料;模型组予生理盐水1 m L/100g灌胃,对照组予多烯磷脂酰胆碱28.5 mg/100 g灌胃,高剂量、中剂量、低剂量组分别予化痰祛湿活血方5.04、2.52、1.26 g/100 g灌胃,连续7周。检测肝功能(ALT、AST、ALP)、血脂(TC、TG)、血糖(GLU)水平,HE染色观察肝组织病理学变化,免疫组化测定ADPN、AdipoR2、p-AKT(ser473)、p-NF-κBp65(ser536)蛋白表达。结果 (1)与空白组比较,模型组大鼠血清ALT、AST、ALP、TC、TG、GLU水平明显升高(P<0.01);与模型组比较,各用药组大鼠血清ALT、AST、ALP、TC、TG、GLU水平均明显下降(P<0.05,P<0.01);与化痰祛湿活血方高剂量组比较,化痰祛湿活血方中、低剂量组及对照组大鼠血清ALT、AST、ALP、TC、TG、GLU水平明显升高(P<0.05,P<0.01)。化痰祛湿活血方高剂量组少数肝细胞胞内有细小脂滴,小叶分界清晰,肝索呈放射状排列;中剂量组部分肝细胞存在大小不等的脂滴,少数汇管区有炎症细胞浸润。(2)与空白组比较,模型组大鼠肝组织ADPN、AdipoR2、p-AKT(ser473)蛋白表达明显下降,p-NF-κBp65(ser536)蛋白表达明显升高(均P<0.01);与模型组比较,各用药组大鼠肝组织AdipoR2、p-AKT(ser473)蛋白表达明显升高,pNF-κBp65蛋白表达明显下降(均P<0.01),化痰祛湿活血方高、中剂量组及对照组大鼠肝组织ADPN蛋白表达明显升高(P<0.01);与高剂量组比较,化痰祛湿活血方中、低剂量组及对照组大鼠肝组织ADPN、AdipoR2蛋白表达明显下降,p-NF-κBp65蛋白表达明显升高(P<0.05,P<0.01)。结论化痰祛湿活血方能通过上调肝组织中ADPN、AdipoR2、p-AKT(ser473)的蛋白表达,抑制p-NF-κBp65(ser536)的蛋白表达,而有效改善肝功能、血脂、GLU,改善NASH大鼠肝组织脂肪变程度,减轻肝脏炎症。
Objective To investigate the therapeutic effects of Huatan Qushi Huoxue Recipe on ADPN, AdipoR2, p-AKT (ser473), p-AKT (ser473) in ADPN / AKT / NF-κB signaling pathway in non-alcoholic steatohepatitis (NASH) Effect of NF-κBp65 (ser536) expression and anti-inflammatory effects. Methods Seventy-two male Sprague-Dawley rats were randomly divided into high-dose, middle-dose and low-dose groups of blank group, model group, control group and Huatan Qushi Huoxue Recipe, with 12 rats in each group. The other five groups were given high-fat diet; the model group was given normal saline 1 m L / 100 g intragastrically, and the control group was given 28.5 mg / 100 g polyene phosphatidylcholine gavage, high dose, middle dose and low dose Respectively phlegm Qushi Huoxue prescription 5.04,2.52,1.26 g / 100 g gavage for 7 weeks. The liver function (ALT, AST, ALP, TC, TG) and blood glucose (GLU) were detected. The pathological changes of liver were observed by HE staining. The expressions of ADPN, AdipoR2, NF-κBp65 (ser536) protein expression. Results Compared with the blank group, the levels of ALT, AST, ALP, TC, TG and GLU in the model group were significantly increased (P <0.01). Compared with the model group, the levels of ALT, AST, (P <0.05, P <0.01). Compared with the high-dose group of Huatan Qushi Huhuo Recipe, the levels of serum ALP, TC, TG and GLU in middle and low dose of Huatan Qushi Huoxue Recipe and control group ALT, AST, ALP, TC, TG, GLU levels were significantly increased (P <0.05, P <0.01). Huatan Huashi Huoxue prescription high-dose group of small hepatocytes intracellular small lipid droplets, leaflets clear boundaries, hepatic cord radial arrangement; part of the medium dose group there are lipid droplets of varying sizes, a small number of portal area inflammatory cell infiltration . (2) Compared with the blank group, the expression of ADPN, AdipoR2 and p-AKT (ser473) in the model group decreased significantly and the expression of p-NF-κBp65 (ser536) Compared with the model group, the expression of AdipoR2 and p-AKT (ser473) and the expression of pNF-κBp65 in the liver of each treatment group were significantly decreased (all P <0.01) (P <0.01). Compared with the high-dose group, the expressions of ADPN and AdipoR2 protein in the liver tissue of middle-dose and low-dose groups of Huatan Qushi Huoxue Decoction group and the control group were significantly decreased , p-NF-κBp65 protein expression was significantly increased (P <0.05, P <0.01). Conclusion Huatan Qushi Huoxue Decoction can effectively improve the hepatic function, blood lipid, GLU, blood lipid, blood lipid and blood lipid level by up-regulating the protein expression of ADPN, AdipoR2 and p-AKT (ser473) in liver tissue and inhibiting the expression of p-NF-κBp65 Improve the degree of hepatic steatosis NASH rats, reduce liver inflammation.