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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 adjuvant therapeutic effect and therapeutic mechanism of apple on experimental hepatic fibrosis in rats. Methods Forty-two male Sprague-Dawley rats were divided into normal group, CCL4 hepatic fibrosis model group, spontaneous recovery group, Dan Shao Huaxian treatment group, Dan Shaohua fiber + apple test group. Rats in the control group were treated with CCL_4 [subcutaneous injection of 40% CCL_4 + peanut oil solution 0.3 mL / 100g for the first time with pure CCL_40.5 mL / 100 g twice daily], and alcohol consumption ), High-fat and low-protein diet (79.5% corn flour, 20% lard, 0.5% cholesterol) and other factors stimulated the preparation of liver fibrosis animal model for 8 weeks. After 8 weeks, 8 rats in the model group were randomly sacrificed, and the rats in the remaining model group were randomly divided into Dan Shao Chemical Fiber (given Danshen chemical fiber powder plus distilled water mixed solution: the drug concentration of 0.1g / 100g rat body weight The rats were fed with 0.2 mL / (10 g · d) gavage once a day and the apple test group (homogenized with fresh apple chopped tissue homogenizer and then filtrated to 4 mL per 100 g of body weight, Fed rats), Danshenhuaxian treatment group and spontaneous recovery group (given normal saline, intragastric administration) for 8 weeks. The degree of hepatic fibrosis was observed by light microscopy after the experiment, and the expression of COL-I and TIMP-1 in the liver was detected by immunohistochemical SP method. Results The expression of collagen-I (COL-I), Tissue inhibitor-of metalloproteinase-1 (TIMP-1) Compared with the model group, the natural recovery group and the Dan-Shao-Hua-Fiber group, the expression was significantly decreased (P <0.01, P <0.05). Conclusion Apple has played an effective role in the treatment of experimental hepatic fibrosis in rats, and its inhibition of lipid peroxidation further inhibits the expression of TIMP-I and COL-I on the basis of the treatment of Dan Shaohua Fibers, thereby promoting the growth of cells External matrix degradation, play a role in the protection of the liver, which may be one of the mechanisms of apple-assisted liver fibrosis reversal.