【摘要】 目的:探讨突变型人肝细胞生长因子(HGF^K132E,R134E,tvNK1)对四氯化碳（CCl₄）诱导的SD大鼠肝纤维化的影响。方法:生物发酵大量制备tvNK1,并经腹腔注射于CCl₄诱导的纤维化SD大鼠体内,6周后取肝脏组织,通过real-time PCR和Western blot检测tvNK1对纤维化SD大鼠肝脏中I型胶原蛋白（Collagen type I,Col1A1）、IV型胶原蛋白（Collagen type IV,Col4A1）和α-平滑肌肌动蛋白（α-smooth muscle actin,α-SMA）在mRNA和蛋白水平表达的影响,并进一步通过HE和Masson染色观察tvNK1对纤维化SD大鼠肝脏形态和肝组织胶原纤维的影响。结果:SDSPAGE检测结果显示,获得纯度≥95%的tvNK1。Real-time PCR和Western blot结果显示,tvNK1降低纤维化大鼠肝脏中Col1A1、Col4A1和α-SMA在mRNA和蛋白水平上的表达。HE和Masson染色结果显示,tvNK1减缓纤维化大鼠肝结构的病理性改变,并降低肝脏中胶原纤维含量。结论:tvNK1能抑制CCl₄诱导的大鼠肝纤维化,为预防肝纤维化和其他器官纤维化疾病提供实验依据。更多还原

【Abstract】 Objective:In order to determine the effects of human truncated variant of hepatocyte growth factor（ tvNK1） on carbon tetrachloride（ CCl₄）-induced liver fibrosis in SD rat.Methods:The recombinant tvNK1 was high-level expression in Escherichia coli BL21 by bio-fermentation and rapid purification of tvNK1 protein using Ni²⁺-affinity chromatograph.The SD rat model of CCl₄-induced hepatic fibrosis was received intraperitoneal injection of tvNK1 for 6 weeks.The mRNA and protein expression levels of Col1A1、Col4A1 andα-SMA in hepatic tissue of different groups were assayed by real-time PCR and Western blot,respectively.The morphology and the collagen fiber of liver sections in different groups were determined by Hematoxylin and eosin（ HE） and Masson staining,respectively.Results:The high purity tvNK1 was assayed by SDS-PAGE.The recombinant tvNK1 reduced the mRNA and protein expression levels of Col1A1、Col4A1 and α-SMA in CCl₄-induced hepatic tissue in Rat.Moreover,injection with tvNK1 restored the morphology and the collagen fiber in CCl₄-induced hepatic tissue in Rat.Conclusion:The recombinant tvNK1 protein attenuated progression of hepatic fibrosis in rat and provided a potential treatment for liver fibrosis and other fibrosis diseases.更多还原