食用苹果辅助逆转大鼠肝纤维化研究中TIMP-1、CoL-Ⅰ的表达

The expression of TIMP-1 and COL-I in experimental study on the adjuvant reverse on rat liver fibrosis

目的研究苹果对实验性肝纤维化大鼠的辅助治疗作用及疗效机理。方法雄性SD大鼠42只分为正常组、CCL_4肝纤维化模型组、自然恢复组、丹芍化纤治疗组、丹芍化纤+苹果试验组。除正常组以外,其余大鼠采用CCL_4[皮下注射40%CCL_4+花生油溶液混合液0.3mL/100g首次用纯CCL_40.5 mL/100 g,2次/周]、饮酒(隔日以30%乙醇为饮料)、高脂低蛋白饮食(79.5%玉米粉,20%猪油,0.5%胆固醇)等复合因素刺激制备肝纤维化动物模型共8周。8周末随机处死模型组大鼠8只,确认造模成功后将剩余模型组大鼠随机分为丹芍化纤[给予丹芍化纤粉末加蒸馏水混合溶液:药物浓度为0.1g/100g大鼠体质量配比,以0.2 mL/(10g·d)灌胃1次]+苹果试验组(新鲜苹果切碎组织匀浆机匀浆后过滤取汁为4mL/100g大鼠体质量灌胃,同时苹果渣做饲料喂养大鼠),丹芍化纤治疗组及自然恢复组(给予生理盐水等量灌胃),共观察8周。实验结束后光镜下观察肝组织纤维化程度,免疫组化SP法检测肝脏中COL-I,TIMP-1的表达。结果免疫组化中丹芍化纤+苹果试验组肝组织中I型胶原(collagen-I,COL-I),基质金属蛋白酶组织抑制因子1(Tissue inhibi-tor of metalloproteinase-1,TIMP-1)的表达与模型组、自然恢复组、丹芍化纤组比较显著降低(P<0.01,P<0.05)。结论苹果在实验性肝纤维化大鼠的治疗中起到了有效的作用,其抑制脂质过氧化作用在丹芍化纤的治疗基础上进一步抑制了TIMP-I、COL-I的表达;从而促进细胞外基质的降解,起到保护肝脏的作用,这可能是苹果辅助逆转肝纤维化的机制之一。

Objective To study the beneficial effects of apple as an adjuvant on rat liver fibrosis and potential mechanisms. Methods Forty-two SD rats were divided into the normal （n=10）, fibrosis model control （n=8）, Danshao Huaxian （DHC） （n=8）, DHC plus Apple and model group （n=8） Liver fibrosis model was induced by complex etiological factors, including CCL4, alcohols （every other day ） and high lipid, low protein diets （79.5% maizena＋ 05% cholesterol ＋20% lard）, for eight weeks. After establishing liver fibrosis, model rats were treated with DHC （0. 1g/（100g·d）, gavage） and DHC （0.1g/（100g·d, garage） plus Apple （apple juice 4mL/100g）, and left untreated （model control ） for another eight weeks. The degree of liver fibrosis was examined under micro- scope. Collagen Ⅰ （COL-Ⅰ ） and Tissue inhibitor of metalloproteinase-1 （TIMP-1） expression were determined by Immunohistochemistry. Results Compared with the DHC, model groups and model control groups, the DHC plus also decreased overexpression of COL- Ⅰ and TIMP-1 in model rat liver （P〈0.01, P〈0.01）. Conclusion Apple has beneficial effects as an adjuvant in the treatment of liver fi- brosis when used together with DHC in rats These effects appear to be related to promote the degradation of extracellular by induction of MMP, and by suppression of COL- Ⅰ and TIMP-1 expression.