参仁活血颗粒对四氯化碳诱导的大鼠肝纤维化TGF-β1/Smad信号通路的影响

Effect of Shenren Huoxue granules on the TGF-β1/Smad signaling pathway in CCl4-induced liver fibrosis in rats

目的观察参仁活血颗粒对四氯化碳诱导的大鼠肝纤维化TGF-β1/Smad信号通路的影响,探讨参仁活血颗粒治疗肝纤维化的作用机制。方法 50只雄性SD大鼠随机分为正常组、模型组和参仁活血颗粒低、中、高剂量组,正常组予以生理盐水1 mL/kg腹腔注射,模型组及参仁活血颗粒各组予以40%四氯化碳0.2 mL/100 g腹腔注射,2次/周,连续8周,于第4周起分别予以参仁活血颗粒,低剂量组每日1.575 g/(kg·bw)、中剂量组每日3.15 g/(kg·bw)、高剂量组每日6.3 g/(kg·bw)剂量灌胃,连续4周,于第8周牺牲大鼠。HE染色及Masson染色检测大鼠肝纤维化程度;免疫组化检测大鼠肝collagen Ⅰ、collagen Ⅲ、TGF-β1的表达;real-time PCR和Western blot检测大鼠肝TGF-β1、Smad3、Smad7的表达。结果模型组与正常组相比可见肝小叶结构破坏,纤维明显增多,形成大小不一的假小叶;而参仁活血颗粒各剂量组较模型组肝纤维化程度明显改善,以参仁活血颗粒高剂量组疗效最显著。②模型组大鼠肝collagen Ⅰ、collagen Ⅲ、TGF-β1、Smad 3表达较正常组明显升高(P<0.05),Smad7表达明显降低(P<0.05),而参仁活血颗粒各剂量组collagen Ⅰ、collagen Ⅲ、TGF-β1、Smad3表达较模型组降低(P<0.05),Smad7表达较模型组升高,以参仁活血颗粒高剂量组疗效最好(P<0.05)。结论参仁活血颗粒能显著改善大鼠肝纤维化,其机制可能与其能下调大鼠肝组织中TGF-β1、Smad3的表达、上调Smad7的表达,从而减少Collagen Ⅰ、Collagen Ⅲ的合成相关。

Objective To observe the effect of a Chinese traditional medicine prescription, Shenren Huoxue(SRHX) granules, on the TGF-β1/Smad signaling pathway in rats with CCl4-induced liver fibrosis and to explore the underlying mechanism. Methods Fifty male SD rats were randomly divided into five groups: control group, model group, low-dose SRHX, middle-dose SRHX and high-dose SRHX groups. The control group was given saline 1 mL/kg;the model group and SRHX granules groups were given 40% carbon tetrachloride(CCl4) 0.2 mL/100 g by intraperitoneal injection twice a week for 8 weeks. From the fourth week, the SRHX groups were given low-dose(1.575 g/(kg·bw)/d), middle-dose(3.15 g/(kg·bw)/d) and high-dose(6.3 g/(kg·bw)/d) SRHX by gastric gavage for 4 weeks and euthanized at week 8. HE staining and Masson staining were used to detect the degree of liver fibrosis in rats. Immunohistochemistry was used to detect the expression of collagen I, collagen Ⅲ and TGF-β1. Real-time PCR and western blot were used to detect the expression of TGF-β1, Smad3 and Smad7 in rat liver tissues. Results Compared with the control group, the liver of the model group showed destruction of liver lobular structure, inflammatory cell infiltration, significantly increased fibers and pseudolobule formation(P< 0.05). Compared with the model group, the degree of liver fibrosis was significantly improved and the high-dose of SRHX granules was the most effective(P< 0.05). The expressions of collagen I, collagen Ⅲ, TGF-β1 and Smad3 in the model group were significantly higher than those in the control group(P< 0.05), while the expression of Smad7 was significantly lower than the control group(P< 0.05). Compared with the model group, the expressions of collagen I, collagen Ⅲ, TGF-β1 and Smad3 in all SRHX groups were decreased while the expression of Smad7 was increased. The high-dose of SRHX granules was better effective(P< 0.05). Conclusions SRHX granules can improve the pathological changes of liver tissue in SD rats with hepatic fibrosis and alleviate the degree of liver fibrosis and liver inflammation in hepatic fibrosis in rats. The mechanism may be related to SRHX granules decreasing the synthesis of collagen I and collagen Ⅲ by decreasing the expression of TGF-β1 and Smad3 and upregulating the expression of Smad7.