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目的:探讨突变型人肝细胞生长因子(HGF~(K132E,R134E),tvNK1)对四氯化碳(CCl_4)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CCl_4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CCl_4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。
Objective: To investigate the effect of mutant human hepatocyte growth factor (HGF ~ (K132E, R134E), tvNK1) on liver fibrosis induced by carbon tetrachloride (CCl_4) in SD rats. Methods: A large amount of tvNK1 was prepared by biofermentation and injected intraperitoneally into CCl 4 -induced fibrosis SD rats. Six weeks later, the liver tissue was taken and the expression of tvNK1 in liver of fibrotic SD rats was detected by real-time PCR and Western blot. Effects of Collagen type I (Col1A1), Collagen type IV (Col4A1) and α-smooth muscle actin (α-SMA) on mRNA and protein expression, and further The effects of tvNK1 on hepatic morphology and hepatic collagen fibers of fibrotic SD rats were observed by HE and Masson staining. Results: The results of SDSPAGE showed that tvNK1 with purity ≥95% was obtained. Real-time PCR and Western blot showed that tvNK1 decreased the mRNA and protein expression of Col1A1, Col4A1 and α-SMA in fibrosis rats. HE and Masson staining showed that tvNK1 attenuated the pathological changes of hepatic structure in fibrotic rats and decreased the content of collagen in the liver. Conclusion: tvNK1 can inhibit CCl_4-induced hepatic fibrosis in rats and provide experimental evidence for preventing liver fibrosis and other organ fibrosis diseases.