膈下逐瘀汤逆转猪血清诱导大鼠肝纤维化的作用及机制研究

Effects of Gexia Zhuyu Decoction on Expression of MMP-13 and TIMP-1 in Rat with Liver Fibrosis

目的:通过观察膈下逐瘀汤对实验性肝纤维化大鼠肝组织基质金属蛋白酶13(matrix metalloproteinase 13,MMP-13)及金属蛋白酶组织抑制因子1(tissue inhibitor of metalloproteinase-1,TIMP-1)的影响,探讨其抗肝纤维化的可能机制。方法:大鼠随机分为空白对照组、膈下逐瘀汤组、猪血清组、猪血清+膈下逐瘀汤组。猪血清2.0 mL.kg-1腹腔注射,每周2次共12周诱导大鼠免疫性肝纤维化模型,同时膈下逐瘀汤每天按7.37 g.kg-1灌胃给药12周。Masson染色法观察肝组织病理学改变,ELISA检测肝组织MMP-13和TIMP-1含量,实时定量Rt-PCR检测肝组织MMP-13和TIMP-1 mRNA表达。结果:猪血清组肝组织胶原面积为(10.38±2.01)%;MMP-13蛋白(14.05±2.64)ng.g-1;MMP-13 mRNA(1.91±0.73)倍;TIMP-1蛋白(14.91±1.07)ng.g-1;TIMP1 mRNA(18.40±2.84)倍。与猪血清组比较,膈下逐瘀汤给药可显著降低肝组织胶原面积(6.66±1.04)%(P<0.05),增加肝组织MMP-13蛋白(20.03±2.14)ng.g-1和MMP-13mRNA(4.12±0.59)倍(P<0.05),降低肝组织TIMP-1蛋白(13.47±1.22)ng.g-1和TIMP-1mRNA(4.89±0.74)倍(P<0.05)。结论:膈下逐瘀汤可有效地逆转实验性肝纤维化大鼠肝组织胶原纤维含量,其机制可能与调节MMP-13和TIMP-1的表达有关。

Objective：To study the effects of Gexia Zhuyu decoction on rat liver fibrosis, and examine its effects on the expression of matrix metalloproteinase-13 （MMP-13） and tissue inhibitors of metalloproteinase-1 （TIMP-1）. Method： Rats were randomly divided into four groups as follows： blank control group, Gexia Zhuyu decoction group, porcine serum group, porcine serum plus Gexia Zhuyu decoction group. The fibrosis model of rats was achieved by intraperitoneal injection of porcine serum twice weekly for 12 weeks, simultaneously Gexia Zhuyu decoction was administered daily by gavage for 12 weeks. All rats were sacrificed at the end of experiment, and liver was collected for analysis. The liver histopathology was examined by Masson trichrome staining. The content of MMP-13 and TIMP-1 in the liver was detected by enzyme linked immunosorbent assay. Real-time reverse transcriptase polymerase chain reaction（Rt-PCR） was used to analyze MMP-13 and TIMP-1 mRNA levels in the liver. Result： Compared with the porcine serum group （10.38±2.01）%;MMP-13 protein（14.05±2.64）ng·g-1 liver;MMP-13 mRNA（1.91±0.73） folds;TIMP-1 protein（14.91±1.07）ng·g-1 liver;TIMP1 mRNA（ 18.40±2.84） folds, the histopathology of rats given Gexia Zhuyu decoction improved significantly （6.66%±1.04%）. The expressions of TIMP-1 mRNA and protein were decreased significantly：protein（13.47±1.22）ng·g-1 liver, and mRNA（4.89±0.74） folds, and MMP-13 mRNA and protein in liver were elevated significantly in porcine serum plus Gexia Zhuyu decoction group：protein（20.03±2.14）ng·g-1 liver; and mRNA（4.12±0.59） folds （P〈0. 05）. Conclusion： Gexia Zhuyu decoction may exert anti-hepatic fibrosis effect in rats via regulation of MMP-13 and TIMP-1 gene expression.