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目的:探讨扶正化瘀方影响基质金属蛋白酶-2活性和Ⅳ型胶原蛋白表达的抗肝纤维化作用机制。方法:二甲基亚硝胺腹腔注射4周诱导大鼠肝纤维化模型,成模后治疗组以扶正化瘀方灌胃4周。检测血清肝功能、肝脏病理学、羟脯氨酸含量;免疫组织化学方法检测肝组织Ⅳ型胶原表达;Western印迹法分析肝组织Ⅳ型胶原、基质金属蛋白酶-2/9(matrix-metalloproteinases-2,MMP-2/9)、金属蛋白酶组织抑制物-2(tissue inhibitors of metalloproteinase-2,TIMP-2)表达水平;明胶酶图法检测肝组织MMP-2/9活性。结果:与正常大鼠比较,模型大鼠血清肝功能异常、肝组织炎症明显、肝组织Ⅳ型胶原表达与沉积增加,TIMP-2蛋白表达增加,MMP-2蛋白表达变化不明显,而MMP-2活性水平明显上升。扶正化瘀方显著减轻肝脏炎症和胶原沉积;减少Ⅳ型胶原蛋白表达和沉积;降低TIMP-2蛋白表达;抑制MMP-2活性。结论:扶正化瘀方可通过抑制肝脏Ⅳ型胶原病理增生沉积、下调MMP-2活性水平、减轻肝组织的破坏与重构而发挥抗肝纤维化的作用。
Objective: To explore the mechanism of Fuzhenghuayu decoction on the activity of matrix metalloproteinase-2 and the expression of type IV collagen in liver fibrosis. METHODS: Rat models of hepatic fibrosis were induced by intraperitoneal injection of dimethylnitrosamine for 4 weeks. After treatment, the treatment group was treated with Fuzheng Huayu Fang for 4 weeks. Serum liver function, hepatic pathology, and hydroxyproline content were detected; immunohistochemistry was used to detect the expression of type IV collagen in liver tissue; Western blot analysis was performed on liver tissue type IV collagen and matrix metalloproteinase-2. , MMP-2/9) and tissue inhibitors of metalloproteinase-2 (TIMP-2) expression were detected by gelatin enzymatic method. RESULTS: Compared with normal rats, the serum hepatic function was abnormal, the inflammation of liver tissue was obvious, the expression and deposition of type IV collagen in liver tissue were increased, the expression of TIMP-2 protein was increased, and the expression of MMP-2 protein was not changed obviously. MMP- 2 The activity level increased significantly. Fuzheng Huayu Recipe significantly reduced hepatic inflammation and collagen deposition; decreased collagen IV expression and deposition; reduced TIMP-2 protein expression; inhibited MMP-2 activity. Conclusion: Fuzheng Huayu Decoction can exert its anti-hepatic fibrosis effect by inhibiting the deposition of liver type IV collagen, down-regulating the activity of MMP-2, and reducing the destruction and remodeling of liver tissue.