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目的探讨鼠白介素6受体融合蛋白(m IL-6RFP)在BALB/c小鼠日本血吸虫感染模型中对肝细胞的保护作用。方法通过亲和层析方法获得可阻断白介素-6(IL-6)的m IL-6RFP,经人类肝癌细胞(Hep G2)IL-6刺激及阻断实验检测纤维蛋白原(fibrinogen,FGG)和结合珠蛋白(haptoglobin,HP)的mRNA表达变化,体外验证重组蛋白m IL-6RFP的生物活性。建立BALB/c小鼠日本血吸虫感染模型,于感染第24天起经尾静脉注射m IL-6RFP,隔天注射每次100μg/只,感染第42天剖杀小鼠。HE染色法检测小鼠肝脏肉芽肿面积,荧光定量PCR法检测小鼠肝组织IL-1β、IL-13、肿瘤坏死因子-α(TNF-α)和趋化因子1(CXCL1)的mRNA表达,连续监测法检测小鼠肝功能指标谷丙转氨酶(ALT)和谷草转氨酶(AST)。结果 Hep G2细胞实验显示m IL-6RFP可降低IL-6对该细胞的刺激作用,显著下调FGG和HP的表达(P<0.05)。在小鼠感染模型中阻断IL-6后,肝功能指标ALT和AST与溶剂对照组相比均显著下降(P<0.05),肉芽肿病变无显著性差异。结论在BALB/c小鼠日本血吸虫感染模型中m IL-6RFP可明显改善肝细胞功能,但对肉芽肿形成未见明显作用。
Objective To investigate the protective effect of murine interleukin-6 receptor fusion protein (mIL-6RFP) on hepatocytes in BALB / c mice infected with Schistosoma japonicum. Methods Interleukin-6 (IL-6) was detected by affinity chromatography and the fibrinogen (FGG) was detected by IL-6 stimulation and blocking assay in Hep G2 cells. And haptoglobin (HP) mRNA expression in vitro, and to verify the biological activity of the recombinant protein mIL-6RFP. The BALB / c mouse model of Schistosoma japonicum infection was established. From the 24th day of infection, m IL-6RFP was injected through the tail vein and 100μg every other day. Mice were sacrificed on the 42nd day of infection. The area of hepatic granuloma in mice was detected by HE staining. The mRNA expressions of IL-1β, IL-13, TNF-α and CXCL1 were detected by real-time quantitative PCR. Continuous monitoring method was used to detect ALT and AST in mice. Results The results of Hep G2 cells showed that mIL-6RFP could reduce the stimulation of IL-6 and significantly down-regulate the expression of FGG and HP (P <0.05). After IL-6 was blocked in mouse model of infection, ALT and AST of liver function decreased significantly compared with that of solvent control group (P <0.05), and granulomatous lesions showed no significant difference. Conclusion m IL-6RFP can significantly improve hepatocyte function in BALB / c mice infected with Schistosoma japonicum, but no significant effect on granuloma formation was found.