相思藤总黄酮对CCl4诱导大鼠肝纤维化的作用研究

Effect of Total Flavonoid of Abrus precatorius on CCl-induced Hepatic Fibrosis Rats

目的观察相思藤总黄酮对CCl4诱导大鼠肝纤维化的保护作用及机制。方法80只SD大鼠随机分为正常组、模型对照组、阳性对照组(秋水仙碱,0.2 mg/kg)、相思藤总黄酮高、中、低剂量组(200,400,800 mg/kg),除正常对照组外,其余各组大鼠灌胃给予50%CCl4花生油混合液制造肝纤维化模型,第8周后,连续4周每日给予大鼠灌胃秋水仙碱和相思藤总黄酮各剂量,正常组和模型对照组灌服等量的生理盐水。末次给药24 h后,取血,收集肝脏组织。检测血清中检测血清丙氨酸转氨酶(ALT),天冬氨酸转氨酶(AST)活性,层黏蛋白(LN),透明质酸(HA),Ⅲ型前胶原(PC-Ⅲ),Ⅳ型胶原(COL-Ⅳ)水平,同时取大鼠肝组织,分别用于苏木素-伊红(HE)染色,MASSON染色观察病理变化,免疫组化法检测转化生长因子-β1(TGF-β1),α-平滑肌肌动蛋白(α-SMA)蛋白表达水平。结果与正常组比较,模型对照组大鼠血清AST,ALT活性及HA,LN,PC-Ⅲ、Col-Ⅳ水平均显著升高,肝组织TGF-β1,α-SMA蛋白表达水平明显增高(P<0.05);与模型对照组比较,相思藤总黄酮能显著降低肝纤维化大鼠血清ALT,AST活性及HA,LN,PC-Ⅲ、COL-Ⅳ(P<0.05)水平,病理组织学检查结果显示,相思藤总黄酮各剂量组大鼠肝组织结构明显改善,肝纤维化程度减轻,相思藤总黄酮高、中剂量组可显著降低肝组织TGF-β1,α-SMA蛋白表达水平(P<0.05)。结论相思藤总黄酮对CCl4诱导的大鼠肝纤维化有明显保护作用,其机制可能与降低肝组织TGF-β1,α-SMA蛋白表达有关。

Objective To observe the protective effect and mechanism of total flavonoid of Abrus precatorius on liver fibrosis induced by CCl4 in rats. Methods Eighty SD rats were randomly divided into normal group,model group,positive control group( colchicine,0. 2 mg/kg),high,medium and low dose of total flavonoid of Abrus precatorius groups( 200,400,800 mg/kg). In addition to the normal control group,the rest groupswere given 50% CCl4 peanut oil mixture by gavage to set up the model of liver fibrosis.After 8 weeks,rats were given colchicine and the total flavonoid of Abrus precatorius with the corresponding dose once a day for continuously four weeks. The normal group and model group took the same amount of normal saline irrigation. Blood and liver tissue were collected 24 h after final administration. The activities of ALT and AST,the serum levels of HA,LN,PC-Ⅲ and COL-Ⅳ were detected. At the same time,the rat liver tissue was taken out for HE staining and MASSON staining to observe the pathological changes. The immunohistochemical method was detected expression levels of TGF-β1 and α-SMA protein. Results Compared with those of the normal group,the model group’s activities of AST and ALT in serum and levels of HA,LN,PC-Ⅲ and Col-Ⅳ were significantly elevated. The expression levels of TGF-β1 and α-SMA protein in the liver tissue increased significantly( P < 0. 05). Compared with those of the model group,the total flavonoids of Abrus precatorius can significantly reduce activities of ALT and AST in serum and the serum levels of HA,LN,PC-Ⅲ and COL-Ⅳ( P < 0. 05). The histologic examination results showed that the structure ofliver tissue in total flavonoids of Abrus precatorius each dose group was obviously improved and the degree of liver fibrosis reduced. The total flavonoids of Abrus precatorius high and middle dose groups can significantly reduce the expression levels of TGF-β1 and α-SMA protein in liver tissue( P < 0. 05). Conclusion The total flavonoids of Abrus precatorius has obvious protective effect on CCl4-induced liver fibrosisrats,and its mechanism may be related to reducing theexpressions of TGF-β1 and α-SMA in liver tissues.