祛湿化瘀方对非酒精性脂肪性肝炎大鼠组织蛋白酶B及肿瘤坏死因子-α表达的影响

[Effects of Qushi Huayu Decoction on cathepsin B and tumor necrosis factor-alpha expression in rats with non-alcoholic steatohepatitis].

作者信息

Zhang Hui, Feng Qin, Li Hong-Shan, Chen Shao-Dong, Wang Xiao-Ning, Peng Jing-Hua, Zhang Ning, Hu Yi-Yang

机构信息

Institute of Liver Diseases, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.

出版信息

Zhong Xi Yi Jie He Xue Bao. 2008 Sep;6(9):928-33. doi: 10.3736/jcim20080910.

DOI:10.3736/jcim20080910

PMID:18782536

Abstract

OBJECTIVE

To study the mechanism of Qushi Huayu Decoction (QHD), a compound of traditional Chinese herbal medicine, in prevention and treatment of non-alcoholic steatohepatitis (NASH).

METHODS

Thirty-five Wistar male rats were randomly divided into normal group, untreated group, QHD group and Ganle (diisopropylamine dichloroacetate) group. The rats except those in normal group were subcutaneously injected with carbon tetrachloride (CCl(4)) for 4 weeks (twice per week) and simultaneously fed with high-fat and low-protein diet for 2 weeks to induce NASH. Then, the rats were administrated with QHD, Ganle, or distilled water for 2 weeks, respectively. After harvest, alanine aminotransferase (ALT) activity and tumor necrosis factor-alpha (TNF-alpha) content in serum as well as triglyceride (TG) and free fatty acid (FFA) in liver tissue were evaluated, and relativity analysis among these parameters was performed. Cathepsin B (Ctsb), phospho-inhibitor kappa B (P-IkappaB), TNF-alpha protein expressions in liver tissue were assayed with western-blot. The expression and distribution of ctsb in liver tissue were observed with immunohistochemical method.

RESULTS

The contents of TG, FFA and activity of ALT were significantly decreased in QHD group. While in the Ganle group, only the activity of ALT in serum was decreased significantly. Expressions of Ctsb, P-IkappaB and TNF-alpha proteins in liver tissues and serum TNF-alpha level were all enhanced in untreated group which, however, were significantly inhibited in the QHD group. And as expected, there were significant relativities among contents of TG in liver tissues and the content of FFA in liver tissue and activity of ALT in serum, content of TNF-alpha in serum and content of FFA in liver tissue and activity of ALT in serum.

CONCLUSION

The inhibiting effects of QHD on fat deposition and inflammation in liver are related with its inhibition on the "FFA-Ctsb-TNF-alpha" pathway of lipo-toxicity.

摘要

目的

研究中药复方祛湿化瘀汤（QHD）防治非酒精性脂肪性肝炎（NASH）的作用机制。

方法

将35只雄性Wistar大鼠随机分为正常组、未治疗组、QHD组和肝乐（二氯醋酸二异丙胺）组。除正常组大鼠外，其余大鼠皮下注射四氯化碳（CCl₄）4周（每周2次），同时给予高脂低蛋白饮食2周以诱导NASH。然后，分别给予大鼠QHD、肝乐或蒸馏水灌胃2周。取材后，检测血清丙氨酸氨基转移酶（ALT）活性、肿瘤坏死因子-α（TNF-α）含量以及肝组织甘油三酯（TG）和游离脂肪酸（FFA）水平，并对这些指标进行相关性分析。采用蛋白质免疫印迹法检测肝组织中组织蛋白酶B（Ctsb）、磷酸化抑制蛋白κB（P-IκB）、TNF-α蛋白表达。采用免疫组织化学方法观察肝组织中Ctsb的表达及分布。

结果

QHD组肝组织TG、FFA含量及ALT活性显著降低。而肝乐组仅血清ALT活性显著降低。未治疗组肝组织Ctsb、P-IκB和TNF-α蛋白表达及血清TNF-α水平均升高，但QHD组显著抑制。且肝组织TG含量与肝组织FFA含量、血清ALT活性、血清TNF-α含量与肝组织FFA含量及血清ALT活性之间均存在显著相关性。