IL-10调节日本血吸虫感染小鼠肝脏炎症和纤维化的实验观察

Experimental observation on IL-10 in regulating liver inflammation and fibrosis in mice infected with Schistosoma japonicum

目的通过日本血吸虫感染小鼠模型,观察IL-10在日本血吸虫感染急性期的作用及其机制,为防治肝纤维化提供新的理论依据。方法采用腹部贴片法建立日本血吸虫感染的小鼠模型。于小鼠感染后第2天开始,实验组注射IL-10R阻断抗体,对照组注射Rat IgG1,每2周注射1次,6周后取血清和肝脏样本检测分析肝脏纤维化相关指标。血清用于检测ALT;肝脏用于HE、天狼星红染色,流式细胞术检测肝脏巨噬细胞亚群变化,定量PCR、ELISA、CBA检测肝脏局部炎症因子变化。结果实验组小鼠较对照组的血清ALT升高(t=5.505,P<0.05),单个虫卵肉芽肿变大(t=4.817,P<0.05),胶原沉积变多,肝脏促炎性Ly6C hi巨噬细胞增多(t=27.63,P<0.05),肝脏局部炎症因子CCL2、CCL3、CCL4、CCL5、TNF-α、IL-1b、IL-6、IL-12a和IL-12b均升高。结论经IL-10R阻断抗体处理的日本血吸虫感染的小鼠表现出更加严重的炎症和纤维化表型,说明IL-10在调节日本血吸虫感染的肝脏炎症和纤维化方面有着极为重要的作用,为今后以IL-10为靶点治疗血吸虫病提供新的思路。

Objective To observe the role and mechanism of IL-10 in the acute phase of Schistosoma japonicum infection in model mice infected by Schistosoma japonicum for new theoretical basis in prevention and treatment of liver fibrosis.Methods Abdominal patch was used to develop mouse models.After two days of infection with Schistosoma japonicum,IL-10R blocking antibody was intraperitoneally applied to the infected mice in experimental group once every two weeks,and Rat IgG1 was administered in mice in control group in the same protocol.By the 6th week,the serum and liver samples were taken to measure the indicators involved in liver fibrosis,including serum ALT level,and pathological changes of the liver after staining with H&E and Sirius red.Flow cytometry was used detect changes of macrophage subgroups in liver tissues,and quantitative PCR,ELISA and CBA were performed to detect the variation of liver inflammatory factors.Results Elevated ALT level(t=5.505,P<0.05),enlarged granuloma in individual egg(t=4.817,P<0.05),increased collagen deposition and count of pro-inflammatory Ly6C hi macrophages in the liver(t=27.63,P<0.05)as well as boosted local inflammatory factors,including CCL2,CCL3,CCL4,CCL5,TNF-α,IL-1b、IL-6、IL-12a and IL-12b,were seen in mice in the experimental group infected with Schistosoma japonicum compared to those in the control group.Conclusion Aggravated inflammation and fibrosis phenotype seen in mice with Schistosoma japonicum infection following IL-10R blocking antibody treatment proves that IL-10 plays an extremely important role in regulating the liver inflammation and fibrosis caused by Schistosoma japonicum,and our findings can provide a new consideration in treating schistosomiasis using IL-10 as the target.