祛痰活血颗粒对NAFLD模型大鼠脂肪组织AQP7，p38MAPK表达的影响

目的:探讨祛痰活血颗粒治疗非酒精性脂肪性肝病(NAFLD)大鼠的作用,并探讨其相关的作用机制。方法:40只SD大鼠随机分成正常组(6只),模型组(34只);除正常组外,采用高脂饮食诱导的方法诱导NAFLD大鼠模型,确认造模成功后,将剩余模型组(30只)随机分为5组,每组6只,分别为模型组,祛痰活血颗粒高、中、低剂量组(10,5,2.5 g·kg~(-1)),SB203580组(3 mg·kg~(-1)),分别予以灌胃祛痰活血颗粒及腹腔注射p38MAPK抑制剂SB203580,连续干预4周。苏木素-伊红(HE)及油红O染色观察肝脏病理变化;试剂盒测定血清甘油三酯(TG),总胆固醇(TC),天门冬氨酸氨基转移酶(AST),丙氨酸氨基转移酶(ALT),肝脏TG;实时荧光定量PCR(Real-time PCR)检测脂肪组织水通道蛋白7(AQP7)及p38分裂原激活的蛋白激酶(p38MAPK)mRNA表达,酶联免疫吸附测定(ELISA)法检测肝脏游离脂肪酸(FFA),脂肪组织AQP7和磷酸化p38MAPK(p-p38MAPK)的表达。结果:与正常组比较,模型组大鼠肝脏脂肪变性明显,血清TG,TC,AST,ALT和肝脏TG,FFA明显升高,脂肪组织中AQP7 mRNA及AQP7含量表达降低,p38MAPK mRNA及p-p38MAPK含量升高(P0.05);与模型组比较,祛痰活血颗粒和SB203580可减轻NAFLD大鼠肝脏脂肪变性程度;明显降低NAFLD大鼠的血清TG,TC,AST,ALT,肝脏TG,FFA(P0.05);各中药组和SB203580组大鼠脂肪组织AQP7 mRNA和AQP7含量表达明显升高,p38MAPK mRNA和pp38MAPK含量明显降低(P0.05)。结论:祛痰活血颗粒能减轻或者逆转肝脏的脂肪变性,其机制可能与抑制脂肪组织p38MAPK活化、上调AQP7表达,增加甘油入血代谢,减少进入肝脏的FFA,从而降低肝脏TG蓄积有关。

Effect of Qutan Huoxue Granules on Expression of AQP7 and p38MAPK in Adipose Tissues of NAFLD Model Rats

Objective: To explore the mechanism of Qutan Huoxue granules in the treatment of non-alcoholic fatty liver disease (NAFLD) in rats. Method: The 40 SD rats were randomly divided into normal group (n=6) and model group (n=34). NAFLD rat models were induced by high fat diet, and after successful modeling, the successful modeling group (n=30) were further randomly divided into 5 groups, 6 in each group:model group, Qutan Huoxue granules high, middle and low dose groups (10, 5, 2.5 g·kg^-1) and SB203580 group (3 mg·kg^-1). Intragastric administration of Qutan Huoxue granules and intraperitoneal injection of p38MAPK inhibitor SB203580 were given respectively for 4 weeks. Then the pathological changes of liver were observed by htoxylin-eosin(HE) staining and oil red O staining; Kits were used to detect the triglyceride (TG), total cholesterol (TC), aspartate transaminase (AST), and alanine transferase (ALT) levels in serum, and TG levels in liver tissues; the adipose tissue aquaporin7 (AQP7) and p38 mitogen activated protein kinase (p38MAPK) mRNA expression levels were detected by Real-time PCR; enzyme linked immunosorbent assay(ELISA) was used to detect the expression of free fatty acid (FFA) in liver, AQP7 and phosphorylated p38MAPK (p-p38MAPK) expression in adipose tissues. Result: As compared with the normal group, the model group showed an obvious hepatic steatosis and a significant increase in serum TG, TC, AST, ALT and liver TG, FFA levels, a significant decrease in the expression of AQP7 mRNA and AQP7 protein and an increase of p38MAPK mRNA and p-p38MAPK protein (P <0.05). As compared with the model group, Qutan Huoxue granules and SB203580 can reduce the degree of liver steatosis, and significantly decrease the serum TG, TC, AST, ALT and liver TG, FFA levels in NAFLD rats (P <0.05). The Chinese medicine groups and SB203580 group showed a significant increase in the expression of AQP7 mRNA and AQP7 protein and a decrease of p38MAPK mRNA and p-p38MAPK protein in adipose (P <0.05). Conclusion: Qutan Huoxue granules can reduce or reverse the liver steatosis, and the mechanism may be related to inhibiting the activation of p38MAPK, up-regulating the expression of AQP7 in adipose tissues, increasing plasma glycerol metabolism, reducing glycerol and FFA intake into liver, hereby reducing TG accumulation in liver.