忍冬木层孔菌总酚通过调控肥大细胞抗四氯化碳所致大鼠肝纤维化

探讨忍冬木层孔菌总酚(Phellinus lonicerinus total phenolics， PLTP)调控肥大细胞(Mast Cell， MC)改善四氯化碳(CCl4)诱导的大鼠肝纤维化的作用.将50只SD大鼠随机分为正常组、模型组、秋水仙碱组(0.2 mg/kg)、PLTP低剂量组(250 mg/kg)、PLTP高剂量组(500 mg/kg)，每组10只.除正常组注射橄榄油外，其余各组腹腔注射2 mL/kg 50% CCl4橄榄油溶液，2次/w，共8 w.首次注射后各给药组每天灌胃对应剂量的药物，正常组与模型组灌胃相应体积的生理盐水，1次/d，共8 w.末次给药后禁食24 h，取大鼠血和肝脏，检测血清中谷丙转氨酶(alanine aminotransferase， ALT)、谷草转氨酶(aspartate aminotransferase， AST)、碱性磷酸酶(alkaline phosphatase， ALP)及透明质酸(haluronic acid， HA)、Ⅲ型前胶原(procollagen typeⅢ， PCⅢ)、Ⅳ型胶原(typeⅣ collagen， Ⅳ-C)的含量；免疫组化测定α-平滑肌肌动蛋白(alpha smooth muscle actin， α-SMA)的表达量，同时切片观察肝脏组织学变化及肥大细胞的数量及脱颗粒变化.结果显示，与模型组比较，PLTP能降低大鼠血清AST、ALT及ALP的活性，降低血清中HA、PCⅢ、Ⅳ-C的含量；病理学检查结果表明PLTP能明显减轻CCl4引起的大鼠肝损伤及纤维化；甲苯胺蓝染色(TB)可见CCl4模型组大鼠肝脏血管周围及纤维间隔内大量正在脱颗粒和已经脱颗粒的充满蓝紫色颗粒的MC，MC数目较正常组明显升高，PLTP组MC数量明显减少.同时，免疫组织化学染色结果显示PLTP下调肝组织中α-SMA的表达.综上，PLTP对CCl4致大鼠肝纤维化有防治作用，其作用机制可能与减少MC的数量及其脱颗粒、下调肝组织中α-SMA的表达有关.

Effect of total phenolics acid of Phellinus lonicerinus (Bond.)Bond. et Sing on hepatic fibrosis by regulating mast cells

To investigate the protective effect of total phenolics acid of Phellinus lonicerinus (Bond.) Bond. et Sing (PLTP) in resisting hepatic fibrosis by regulating mast cells. 50 rats were randomly divided into normal group， model group， colchicine group (0.2 mg/kg) ， PLTP-high and low dose group(500， 250 mg/kg). Rat hepatic fibrosis was induced by intraperitoneal injection of 50% CCl4-olive oil solution at a dose of 2 mL/kg twice weekly for 8 w. After the first injection， the normal group and model group were given equivoluminal physiological saline， and other groups were given the corresponding does of drugs once per day for 8 w. After the last administration， the rats were fasting for 24 h， and all the rats were killed with blood samples and liver tissues collected The serum contents of alanine aminotransferase (ALT)， aspartate aminotransferase (AST)， alkaline phosphatase (ALP)， hyaluronic acid (HA)， precollagen type Ⅲ (PCⅢ) and typeⅣ collagen (Ⅳ-C) were measured biochemical method. The α-smooth muscle actin (α-SMA) were detected by immunohistochemical assays. Meanwhile， the histopathological changes and mast cell (MC) of the liver tissue were observed. The results showed that the PLTP could significantly improve the liver tissue lesion， decrease the levels of ALT， AST， ALP， HA， PCⅢ and Ⅳ-C. The microexamination results of histological sections showed that PLTP significantly reduced the rat liver injury and fibrosis induced by CCl4. The CCl4 model group exhibited flatty degeneration and fibrosis with many degranulating and degranulated mast cells filled with purple granula located around the liver blood vessels and in fiber-interval， while those of PLTP-treated group decreased significantly. Immunohistochemical staining suggested that the expression of α-SMA was down-regulated with PLTP treatment. In summary， the PLTP possessed preventive and therapeutic effects on hepatic fibrosis induced by CCl4 in rats， and the mechanism might be related to the decrease of the number of mast cell and the expression of α-SMA in liver tissues.