【作者】 王伟芹；

【导师】 尹常健；

【作者基本信息】 山东中医药大学 ， 中西医结合临床， 2007， 博士

【摘要】 目的:观察清肝调脂饮对非酒精性脂肪性肝炎的临床疗效及优势,并通过动物实验探讨其作用机制。方法:临床观察:以70例非酒精性脂肪性肝炎患者为研究对象,随机分为清肝调脂饮治疗组和西利宾胺对照组,每治疗4周为一个疗程,8周为一总疗程。观察两组患者治疗过程中的临床证候、血生化指标(ALT、AST、γ-GT、TC、TG)和肝脏B超的改善情况。实验研究:Wistar大鼠70只,除10只作为正常对照组外,余60只建立以高脂饮食(基础饲料88%,猪油10%,胆固醇1.5%,胆盐0.5%)诱导的非酒精性脂肪性肝炎大鼠模型并随机分组,除模型组外,清肝调脂饮低、中、高剂量组和西利宾胺组分别灌胃相应剂量药物治疗。实验12周后,各组大鼠全部处死,测量大鼠体重及身长,计算Lee’s指数;检测各组大鼠血清肝功能指标(ALT、AST、ALP)、血脂指标(TC、TG、FFA)以及血清瘦素(Leptin),测定空腹血糖(FBG)和空腹胰岛素(FINS)并计算胰岛素抵抗指数(IRI)和胰岛素敏感指数(ISI);检测各组大鼠肝组织匀浆肿瘤坏死因子(TNFα)和MDA含量以及SOD活性;各组大鼠肝组织行HE染色,Bcl-2、CD14、CYP2E1免疫组化染色以及特殊染色(糖原,脂肪及含铁血黄素染色),阳性结果作定量分析。结果:临床数据经统计学处理后发现,与治疗前比较,治疗后两组患者各观察指标均有不同程度的改善(P<0.05,P<0.01),其中治疗组临床综合疗效明显优于对照组(P<0.05)。对各指标改善情况比较发现,两组除肝功能指标改善无差异外(P>0.05),治疗组临床证候、血脂以及肝脏B超的改善情况均不同程度优于对照组(P<0.05,P<0.01)。实验结果经统计学处理后发现,与空白对照组比较,模型组以上各指标均显示异常(P<0.05,P<0.01);而与模型组比较,各用药组均能不同程度的改善以上各指标(P<0.05,P<0.01),其中,在改善TG、FFA、FBG、INS、IRI、ISI、TNFα方面,中药高剂量组均明显优于西药对照组(P<0.05,P<0.01);而对其他各指标的改善,中药各剂量组与西药对照组疗效相似,但前者有优于后者的趋势。结论:清肝调脂饮防治非酒精性脂肪性肝炎的疗效机制包括:1.清肝调脂饮主要通过改善胰岛素抵抗和瘦素抵抗等,促进游离脂肪酸代谢和肝内TG分解,增加肝糖原合成,从而纠正血脂紊乱,减少脂肪在肝细胞内沉积;2.清肝调脂饮通过降低脂质过氧化反应,提高抗氧化能力,减少细胞色素P450ⅡE1的活性表达,减轻肝组织炎性细胞浸润,抑制肝脏库普否细胞的活化,减少细胞因子尤其是TNFα的释放,抑制肝细胞凋亡等,从而减轻肝细胞变性坏死程度,促进肝细胞修复。因此,清肝调脂饮主要通过以上作用途径,使非酒精性脂肪性肝炎患者的临床证候、血生化指标以及肝脏的脂肪肝程度均得到不同程度的改善。与西利宾胺比较,清肝调脂饮在抑制非酒精性脂肪性肝炎的始动因素方面具有明显的疗效优势。更多还原

【Abstract】 Objective:To observe the clinical curative effect and advantage of Qinggantiaozhi Decoction on nonalcoholic steatohepatitis, and explore the mechanism of it by animal experiment.Methods:Clinical observation: 70 patients with nonalcoholic steatohepatitis (NASH) were divided into two groups in random: the treated group with Qinggantiaozhi Decoction and the control group with silybin. It was 4 weeks for a course of treatment, and the total course of treatment was 8 weeks. In the course of treatment, the items that we had observed with all patients were sings and symptoms, biochemical indexes including alanine aminotransferase (ALT), aspartate aminotransferase (AST),γglutamyltransferase (γ-GT), total cholesterol (TC) and triglyceride (TG), and the B-type ultrasonic inspection of liver. Empirical study: 70 Wistar rats, and 10 rats were divided to the normal control group, and the other 60 rats were induced to the model of NASH with high fat diet that was made up based animal feed (88%), lard (10%), cholesterol (1.5%) and bile salt (0.5%). The 60 rats were divided into five groups in average: model groups, the groups of Qinggantiaozhi Decoction in low dose, middle dose and high dose, and the contrast group of silybin. In the five groups, except the model group, others were treated by corresponding medicine. After 12 weeks, all rats were put to death, and the following items were detected: the index of Lee’s through measuring the body weight and length of rats; the serum levels of ALT, AST and alkaline phosphatase (ALP); the serum contents of TC, TG, free fatty acids (FFA) and leptin; the serum contents of fasting blood glucose (FBG) and fasting insulin (FINS), and to calculate the insulin resistant index (IRI) and the insulin sensitive index (ISI) with them. The contents of tumor necrosis factor-alpha (TNFα) and malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) were detected in the liver tissue homogenate. The liver histopathology was observed by hematoxylin and eosin (HE) staining. The B cell lymphoma/leukemia-2 (Bcl-2), CD14 and cytochrome P450ⅡE1 (CYP2E1) were expressed on the liver tissue by immunohistochemistry staining, the amylon, fat and hemosiderin were expressed on the liver tissue by specific staining. All a b o v e p o s i t i v e r e s u l t s w e r e a n a l y z e d b y q u a n t i t a t i v e a n a l y s i s .Results:After all clinical data were counted with statistics, we found that, to compare with prior treatment, in post treatment all indexes in the patients of two groups are improved on different degree (P<0.05,P<0.01), and the clinical synthetic curative effect in treated group is better than that in contrast group (P<0.05). To the improvement of indexes of liver function, there is no difference between two groups (P>0.05), but the clinical sings and symptoms, contents of blood-fat and the manifestation of B-type ultrasonic inspection in liver are all improved better in treated group than those in contrast group (P<0.05,P<0.01). After to analyze the experimental data, we found that, all above indexes in model group are abnormal compared with those in normal group (P<0.05,P<0.01), but the indexes are all improved in treated groups than those in model group (P<0.05,P<0.01). The indexes of TG, FFA, FBG, INS, IRI, ISI and TNFαare all improved better in high dose group of Qinggantiaozhi Decoction than those in silybin group (P<0.05,P<0.01), and for others, there are similar curative effects between Qinggantiaozhi Decoction groups and silybin group, however, there is still a tendency that curative effect in Qinggantiaozhi Decoction groups is better than that in silybin group.Conclusion:There are several mechanisms on Qinggantiaozhi Decoction preventing and curing NASH. The first one is to correct disordered blood-fat and reduce the deposition of fat in hepatic cell through improving the resistance of insulin and leptin, promoting metabolism of FFA and TG in hepatic cell. The second one is to lessen hepatic cell degeneration and to promote cytothesis through depressing the lipid peroxidatic reaction, elevating antioxygen, decreasing the active expression of CYP2E1, lessening inflammatory cell to infiltrate, restraining the activation of kupffer, reducing the release of cytokine especial TNFαand suppressing apoptosis of hepatic cell. Based on above mechanisms, Qinggantiaozhi Decoction can improve sings and symptoms, serum biochemical indexes and the display of fatty liver of patients with NASH, and compared with silybin, Qinggantiaozhi Decoction have better curative effect to restrain the initiating agent of NASH.更多还原