黄芪对肝纤维化的防治作用(1)
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[摘要] 目的:观察黄芪的抗肝纤维化作用,初步探讨黄芪抗肝纤维化的机制。方法:设立正常对照组、模型组和黄芪处理组,以四氯化碳(CCl4)皮下注射复制大鼠肝纤维化模型,黄芪处理组在造模的同时给予皮下注射黄芪注射液。常规HE染色观察肝脏病变,天狼猩红胶原染色、肝组织羟脯氨酸(HYP)含量测定观察肝纤维化程度;产色基质偶氮法鲎试剂定量测定血浆内毒素,赖氏法测定血浆丙氨酸氨基转移酶(ALT);TBA法检测肝组织丙二醛(MDA)水平。结果:黄芪处理组与模型组比较,①肝脏的损伤性改变有所减轻,肝组织HYP及肝纤维化指数(FI)明显降低;②血浆内毒素含量、ALT及肝组织MDA均有不同程度的降低。结论:黄芪可减缓肠源性内毒素血症,可能为防治肝纤维化的作用机制之一。
[关键词] 内毒素;肝纤维化;黄芪
[中图分类号] R285.5[文献标识码]A [文章编号]1673-7210(2010)03(c)-015-03
Inhibitive effect of Astragalus on hepatic fibrosis
WANG Dengni1, XU Junquan1*, SONG Weifang1, ZHANG Xiaoyang2, WANG Mingliang1, LIANG Yonggang1
(1.Science and Technology Center, Fenyang College of Shanxi Medical University, Fenyang 032200, China; 2.Department of Pathology, the Second Hospital of Tianjin Medical University, Tianjin 300211, China)
[Abstract] Objective: To observe the action of Astragalus on anti-hepatic fibrosis, in order to probe into the mechanism of Astragalus in resisting hepatic fibrosis. Methods: Normal control group, model group and Astragalus treatment group were set in the experiment, model of hepatic fibrosis was induced by subcutaneous injection of carbon tetrachloride (CCl4). Astragalus treatment group was given subcutaneous Astragalus when establishing models. Liver tissues were stained with hematoxylin-erosin to observe liver injury. The degree of liver fibrosis was observed by stained of sirius red of collagen and the content of hydroxyproline in liver tissues. Endotoxin was measured with a limulus amebocyte lysate test kit. Plasma alanine aminotransferase (ALT) was determined with Reitman-Frankel method; the content of malondialdehyde (MDA) in liver tissue was determined with TBA method. Results: Astragalus treatment group compared with model group: ①changes in liver damage were relieved, HYP in liver tissue and hepatic fibrosis index (FI) were significantly reduced; ②plasma endotoxin, ALT and liver MDA were reduced to different degrees. Conclusion: Astragalus can alleviate CCl4-induced liver injury, stave happening and developing of hepatic fibrosis; Astragalus can reduce intestinal endotoxemia, which may be one of the mechanisms contributing to the prevention and treatment of Astragalus on liver fibrosis.
[Key words] Endotoxin; Hepatic fibrosis; Astragalus
在抗肝纤维化的众多药物中,中药黄芪及其复方制剂是一类有很大发展潜力的药物。以往的动物实验结果显示,黄芪具有抑制肝组织胶原沉积和防治肝纤维化的作用,还可改善肝脏微循环,增强网状内皮系统的吞噬功能,促进肝再生。然而有关其抗肝纤维化的机制还有待进一步研究。内毒素可激活肝枯否细胞(Kupffer cell,KC)合成与释放细胞因子(cytokine)和炎症介质,并在肝纤维化的发生及发展中发挥作用,内毒素通过诱导TNF-α产生[1],促进成纤维细胞和间质细胞增殖和分化,使细胞外基质(extracellular matrix,ECM)产生增多。本研究通过观察黄芪对慢性肝纤维化过程中血浆内毒素水平的影响及意义,进一步探讨黄芪防治肝纤维化的作用机制。
1材料与方法
1.1动物及材料
健康清洁级雌性Wistar大鼠,体重(230±20) g,购自山西医科大学实验动物中心;四氯化碳(CCl4)为中国中邦化工分析纯;鲎试剂盒购自上海伊华医学科技公司,天狼猩红、固绿FCF、羟脯氨酸(HYP)标准品购自美国Sigma公司;黄芪注射液(成都地奥九鸿制药厂),丙二醛(MDA)测定试剂盒(南京建成生物工程研究所第一分所),丙氨酸氨基转移酶(ALT)试剂盒(北京北化康泰临床试剂有限公司)。
1.2 方法
1.2.1 分组30只Wistar大鼠饲养1周后随机分为三组,A组(正常对照组)、B组(模型组6周)、C组(黄芪处理组6周)。
1.2.2 造模A组不给予处理,其他两组给予首剂5 ml/kg体重皮下注射CCl4分析纯,然后按3 ml/kg体重皮下注射40% CCl4植物油(V/V)混合液,每周2次。C组于造模同时给予皮下注射黄芪注射液[800 mg/(kg·d)][2]。
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