【作者】 江宇泳；

【导师】 王宪波；

【作者基本信息】 北京中医药大学 ， 中西医结合临床， 2009， 硕士

【摘要】 目的:1.观察八味抗纤方对四氯化碳CCl₄诱导的肝纤维化大鼠的治疗作用,并初步探讨其可能机制。2.观察八味抗纤方治疗慢性乙型肝炎肝纤维化的临床疗效。方法:1.采用四氯化碳橄榄油溶液腹腔注射,每周2次,共8周,制作大鼠肝纤维化模型,自造模之日起,分别给予中药高剂量、低剂量灌胃,观察高、低剂量的八味抗纤方对慢性肝损伤大鼠血清中ALT、AST含量的影响,采用Ishak肝病组织病变计分修正方案,结合H.E.染色、Masson染色和免疫组化染色研究八味抗纤方对肝纤维化大鼠肝组织病理改变的影响,以活化HSC特异性标记α-SMA标记显示活化HSC,以探讨八位抗纤方抗肝纤维化的可能的作用机制。2.将66例中医辨证为气虚血瘀,湿热未尽的慢性乙型肝炎患者按2:1比例随机分为中药治疗组和对照组,中药治疗组44例,对照组22例,两组患者均给予基本保肝治疗:维生素C片2粒,每日3次+复合维生素B片2粒,每日3次,其中中药治疗组加用八味抗纤方（黄芪、丹参、党参、白术、郁金、黄芩、陈皮、石菖蒲）,每日300ml,早晚分服,疗程6个月。观察两组患者中医症状变化、肝功能、血清肝纤维化指标（透明质酸、Ⅲ型前胶原肽、Ⅳ型胶原、层粘连蛋白）等,结合部分病例治疗前后肝组织活检对比,观察八味抗纤方对慢性乙型肝炎肝纤维化的治疗作用。结果:1.肝纤维化模型大鼠的血清ALT、AST和TBIL均显著高于正常组,中药低剂量组和高剂量组的AST均显著低于模型组;模型组大鼠肝细胞气球样变性,脂肪变性明显,见到较多的嗜酸性变性及,Masson染色显示肝小叶结构紊乱,汇管区扩大明显,胶原沉积明显增多,纤维间隔较宽,胶原致密,甚至见到假小叶形成;肝组织α-SMA阳性染色区域较宽,着色强烈,汇管区、纤维间隔区、肝窦均有大量表达,纤维间隔区阳性染色呈条带状分布,粗细不均,随肝纤维化程度加重而增多。中药低剂量组、中药高剂量组肝细胞气球样变性,脂肪变性、嗜酸性变性均少于模型组,也有纤维组织增生,程度较轻,纤维间隔较少、较细;α-SMA阳性细胞数少于模型组,部分阳性染色呈细束状分布。根据Ishak肝病组织病变计分修正方案计分,中药低剂量组、中药高剂量组炎症和纤维化程度明显轻于模型组,三组间炎症和纤维化评分均有显著性差异。2.两组慢性乙型肝炎患者治疗前在年龄、肝功能、血清肝纤维化指标、肝组织病变等方面具有良好可比性。中药治疗组乏力症状显著改善,对血清肝纤维化指标的联合评价显示中药治疗组显著优于对照组（P=0.020）,中药尤其可以明显改善患者血清HA的异常升高。18例患者完成治疗前后的二次肝活检,治疗组纤维化分期较治疗前下降的2例,8例无变化,较治疗前升高的1例;对照组4例患者纤维化分期与治疗前相比无变化,3例较治疗前升高。结论:八味抗纤方能够抑制CCl₄诱导的大鼠肝纤维化,抑制纤维结缔组织的增生和假小叶的形成。其作用机制可能是:保护肝细胞,抑制肝星状细胞的活化及促进细胞外基质降解。慢性乙型肝炎肝纤维化的基本病机是气虚血瘀,湿热疫毒残留难尽。中药八味抗纤方具有益气活血,清化湿热,化痰散结的功效,祛邪不伤正,化瘀而生新,能有效改善血清肝纤维化指标,治疗前后肝活检病理组织学变化显示能阻止肝纤维化的进展。更多还原

【Abstract】 Objective:To observe therapeutic effect of Eight-Ingredient Decoction of Anti-Fibrosis against experimental liver fibrosis induced by carbon tetrachloride（CCl₄）,and to discuss its therapeutic mechanism initially.To observe the clinical effect of the Eight-Ingredient Decoction of Anti-Fibrosis treated liver fibrosis in chronic hepatitis B.Method:1.The carbon tetrachloride mixed with olive oil was injected intraperitoneal for 16 times to establish the model of the rat with liver fibrosis.Since the beginning of the model establishment,the rats were administrated with high dose of TCM.and low dose of TCM separately.Observe the influence of ALT and AST of the rats with ’hepatic injuries. Referring to of Ishak liver disease pathological changes correction scoring scheme,make use of HE stain,Masson stain and immunohistochemistry stain to study the influence of the Eight-Ingredient Decoction of Anti-Fibrosis against rats’ liver fibrosis.Make use of a-SMA immunohistochemistry stain marked the activating HSC to discuss the probably mechanism of Eight-Ingredient Decoction of Anti-Fibrosis against liver fibrosis.2.Sixty-six patients with chronic hepatitis B who are differentiated with Qi deficiency and blood stasis along with Dampness and Heat are randomly separated into TCM group and control group by ratio of 2:1,with 44 patients in TCM group and 22 patients in control group.The patients in both group were treated with basic liver protection treatment which is made of 2 vitamin C pills t.i.d and vitamin B complex pills t.i.d.The TCM group was also treated with Eight-Ingredient Decoction of Anti-Fibrosis which consists of huangqi, danshen,dangshen,baizhu,yujin,huangqin,chenpi,shichangpu,300ml a day,patients drink the liquor in the morning and at night for six months.To observe the TCM symptoms, liver function,mark of liver fibrosis which is made up with hyaluronic acid,precollagenⅢ,Ⅳtype collagen,laminin.Compared to the liver issue biopsy of some cases before the treatment,we observe the therapeutic action of Eight-Ingredient Decoction of Anti-Fibrosis against liver fibrosis of chronic hepatitis B.Results:1.The level of ALT,AST,TBIL of model of the rats with liver fibrosis are significantly higher than the normal group.The AST level of both group with high TCM dose and low TCM dose are all significant lower than the model control group.Ballooning degeneration and fatty degeneration of liver cells can be seen very clear in the model group, which has more acidophilic degeneration and apoptotic body.Masson stain shows loss of normal structure of hepatic lobules,portal areas is enlarged,collagen deposition is increased,the fibrous septa is broadened,collagen is compact and even pseudolobule proliferation in the model group.Theα-SMA positive stain region of liver issue is broad and significantly stained along with portal areas,the fibrous septa and hepatic sinusoid,the positive stain of the portal areas shows zonal distribution and the width odds,and increased by the level of liver fibrosis.While in the TCM group,Ballooning degeneration and fatty degeneration of liver cells,acidophilic degeneration and apoptotic body are all fewer than the model group.There are few fibrous tissue and hyperplasia with lower degrees,the fibrous septa are fewer and finer.The number ofα-SMA positive cells is smaller,which shows fine bundles distribution.According to pathological changes of liver disease correction scoring scheme,both level of inflammation and fibrosis of the group with high TCM dose and low TCM dose are significantly lower than that of the model control group. The score of level of inflammation and fibrosis is significantly different.2.The two group patients of chronic hepatitis B is comparable in age,liver function, multiply serum marks and fibrosis lever of liver biopsy.The patients’ symptom of hypodynamia is remarkable improved in TCM treatment group.The TCM treatment group shows marked effect on the basis of multiply serum marks of fibrosis compared to the control group（P=0.020）.TCM can improve the abnormally increased of HA.Eighteen patients completed both the liver biopsy before and after the treatment.Two of the patients’ lever of fibrosis in the treatment group declined,seven cases remained the same,while only one case ascends.At the same time,five patients’ lever of fibrosis in the control group remained the same,and three cases ascend.Conclusion:Eight-Ingredient Decoction of Anti-Fibrosis can inhibit rat liver fibrosis induced by CCl₄,fiber connective tissue hyperplasia and formation of pseudo-lobe.The mechanism of it is likely to be inhibition of hepatic stellate cell activation and collagen synthesis to protect hepatic cells.The basic pathogenesis of chronic hepatitis with liver fibrosis is qi deficiency and blood stasis,and phlegm-damp obstruction in collaterals.This decoction has the function of replenishing qi and activating blood,clearing heat and resolving dampness,resolving phlegm and dissipating mass,which can make vital qi sufficient,circulation of qi and blood fluent,treat both root and manifestation and interrupt live fibrosis.更多还原