Department of Pharmacology, Tianjin Medical University, No. 22, Qixiangtai Road, Heping District, Tianjin 300070, China.
World J Gastroenterol. 2010 Mar 28;16(12):1458-64. doi: 10.3748/wjg.v16.i12.1458.
To explore the anti-fibrotic effect of Haobie Yangyin Ruanjian decoction (HYRD) on CCl(4)-induced hepatic fibrosis in rats and its modulation on the transforming growth factor (TGF) beta-Smad signaling pathway.
Fifty-six healthy Wistar rats were randomly divided into five groups: normal control group (n = 6), CCl(4)-induced hepatic fibrosis group (n = 14) and three treatment groups (the treated rats received HYRD via oral administration at daily dosages of 8.2, 2.5 and 0.82 g/kg, respectively) of HYRD (n = 12, respectively). Experimental hepatic fibrosis was induced by subcutaneous injection of carbon tetrachloride solution (CCl(4) dissolved in peanut oil, 4:6, V/V) with 0.5 mL/100 g body weight for the first time, and then 0.3 mL/100 g body weight twice a week for 8 wk. In the former 2 wk, rats were raised by feedstuff I (80% corn meal, 20% lard, 0.5% cholesterol). After 2 wk, they were raised by feedstuff II (corn meal and 0.5% cholesterol). Except for the control group, 30% alcohol solution was given orally to each rat every other day from the beginning, 1 mL for each rat. Liver function parameters and hepatic hydroxyproline content were detected by chromatometry. Serum levels of hyaluronic acid (HA), type IV collagen (CIV), type III precollagen (PCIII) and laminin (LN) were assayed with radioimmunoassay. Deposition of collagen was observed with hematoxylin-eosin staining and collagen staining. Gene expression of TGFbeta1 and Smad3 were detected with real-time reverse transcriptase-polymerase chain reaction and Western blotting, respectively.
The serum levels of alanine transaminase and aspartate transaminase were increased in the model group compared with the control group (P < 0.01), and they were decreased in the three treatment groups compared with the model group. The serum levels of total protein and albumin were decreased in the model group and increased in the three treatment groups. The hepatic hydroxyproline content and serum levels of PCIII, HA, LN and CIV were markedly increased in the model group compared with the control group, and decreased in the treatment groups. The gene expression of TGFbeta1 and Smad3 was enhanced in the model group compared with the control group, and HYRD could down regulate their expression.
HYRD can inhibit hepatic fibrosis induced by CCl(4) in rats, which is probably associated with its down-regulation on fibrogenic signal transduction of TGFbeta-Smad pathway.
探讨蒿鳖养阴软坚汤（HYRD）对 CCl4 诱导的大鼠肝纤维化的抗纤维化作用及其对转化生长因子（TGF）β-Smad 信号通路的调节作用。
56 只健康 Wistar 大鼠随机分为 5 组：正常对照组（n=6）、CCl4 诱导的肝纤维化组（n=14）和 3 个治疗组（分别给予 HYRD 灌胃，剂量为 8.2、2.5 和 0.82 g/kg/d，n=12）。采用皮下注射四氯化碳溶液（溶于花生油的四氯化碳，4:6，V/V）0.5 mL/100 g 体重首次，然后每周两次 0.3 mL/100 g 体重 8 周，诱导实验性肝纤维化。在前 2 周，大鼠用饲料 I（80%玉米粉、20%猪油、0.5%胆固醇）饲养。2 周后，它们用玉米粉和 0.5%胆固醇的饲料 II 饲养。除对照组外，从第 1 天开始，每天给每只大鼠口服 30%酒精溶液 1 mL。用色谱法检测肝功能参数和肝羟脯氨酸含量。用放射免疫法检测血清透明质酸（HA）、IV 型胶原（CIV）、III 型前胶原（PCIII）和层粘连蛋白（LN）水平。用苏木精-伊红染色和胶原染色观察胶原沉积。用实时逆转录-聚合酶链反应和 Western 印迹法分别检测 TGFβ1 和 Smad3 的基因表达。
与对照组相比，模型组大鼠血清丙氨酸转氨酶和天冬氨酸转氨酶水平升高（P<0.01），3 个治疗组大鼠血清丙氨酸转氨酶和天冬氨酸转氨酶水平均低于模型组。与模型组相比，3 个治疗组大鼠血清总蛋白和白蛋白水平降低。与对照组相比，模型组大鼠肝羟脯氨酸含量及血清 PCIII、HA、LN 和 CIV 水平显著升高，治疗组大鼠肝羟脯氨酸含量及血清 PCIII、HA、LN 和 CIV 水平降低。与对照组相比，模型组大鼠 TGFβ1 和 Smad3 基因表达增强，HYRD 可下调其表达。
HYRD 可抑制 CCl4 诱导的大鼠肝纤维化，可能与其下调 TGFβ-Smad 通路的纤维生成信号转导有关。
