川芎嗪对大鼠肝纤维化转化生长因子-β_1/Smads信号通路的影响

Effects of ligustrazine on transforming growth factor beta-β_1/Smads signal pathway of liver fibrosis in rats

目的研究川芎嗪对实验性大鼠肝纤维化的治疗作用,并探讨其对转化生长因子(TGF)-β_1/Smads信号通路可能的影响。方法80只雄性SD大鼠,随机挑选70只予皮下注射40%四氯化碳,其余10只作为正常对照组(N组),8周后造模组中随机处死16只(M组)并证实肝纤维化形成,其余肝纤维化大鼠随机分成2组,分别予以川芎嗪腹腔注射(T组,80mg·kg^(-1)·d^(-1))和0.9%氯化钠溶液腹腔注射(R组),疗程为8周。实验结束所有大鼠采血后处死,分别行苏木精-伊红(H-E)染色,采用纤维化半定量计分系统评估肝纤维化程度,Masson染色评估肝组织中胶原纤维百分比,荧光定量聚含酶链反应检测肝组织内TGFβ_1、Smad3和Smad7表达。结果①T组大鼠肝纤维化程度和肝组织中胶原面积密度分别为7.8±2.5和11.68±2.26,较R组的10.2±2.8和18.84±2.74明显减轻(P值分别<0.05、0.01)。②M组大鼠肝组织中TGF-β_1、Smad3的相对表达分别为1.54±0.08和1.62±0.03,较N组的0.78±0.15和0.88±0.17明显升高(P值均<0.01),M组Smad7相对表达为0.88±0.11,显著低于N组的1.31±0.02(P<0.01)。③T组肝组织TGF-β_1、Smad3分别1.02±0.09和1.07±0.01,均显著低于M组(P值均<0.01),而Smad7为1.15±0.01,显著高于M组(P<0.01)。结论川芎嗪具有明显的逆转肝纤维化作用,其机制可能与其降低TGF-β_1的表达以及改善Smad3与Smad7之间的失衡有关。

Objective To investigate the therapeutic effects of ligustrazine on experimental hepatic fibrosis in rats, and to appraise the effects on TGF-β1/Smads signal pathway. Methods Hepatic fibrosis was induced by subcutaneous injection of carbon tetrachloride in 70 male SD rats（twice a week, 3 mL/kg）within 8 weeks and another 10 rats were routinely fed as normal group （N group）. 16 rats were sacrificed to ensure the presence of hepatic fibrosis （M group）, then the rest rats with hepatic fibrosis were randomly devided into two groups receiving intraperitoneal injection of ligustrazine （80 mg · kg^-1 · d^-1 , T group） and the same dosage of normal saline（R group）for 8 weeks. The staging of hepatic fibrosis was assessed through HE staining and observed under light .microscope（semi quantitative scaling system, SSS）. The area density of collagen in liver speciments was assessed through Masson staining; and the expressiofi of TGF-β1 , Smad3 and Smad7 in hepatic tissue were assessed by fluorescence quantitative polymerase chain reaction （PCR）. Results （1）The staging of hepatic fibrosis in T group was obviously reduced when compared with R group （7.8 ± 2.5 vs 10.2 ±2.8; P 〈 0.05） ; the area density of collagen in liver tissues was obviously lower in T group（11. 68 ± 2. 26 vs 18.84 ± 2. 74, P 〈 0.01）. （2）The expression of TGF-β1 and Smad3 were markly elevated in M group than those in N group （1.54 ± 0.08 vs 0. 78 0.15, 1.62 ± 0.03 vs 0. 88 ± 0.17, respectively ; P 〈 0.01）, the expression of Smad7 in M group was reduced （0.88 ± 0.11 vs 1.31 ± 0.02, P 〈 0.01） . （3）Compared to M group, the expression of TGF-β1 and Smad3 in Tgroup was markly decreased（1.02 ± 0.09 vs 1.54 ± 0.08, 1.07 ± 0.01 vs 1.62 ± 0.03, respectively; P 〈 0.01）, whereas the Smad7 was increased （ 1.15 ± 0.01 vs 0. 88 4- 0.11 ; P 〈 0.01）. Conclusion Ligustrazine has obvious therapeutic effect on hepatic fibrosis in rats. The mechanism may be correlated with reduced expression of TGF-β1 and improvement of the disequilibrium of Smads3 and SmadsT.