CY-09减轻血吸虫病肝纤维化的作用研究

CY-09 in reducing liver fibrosis caused by schistosomiasis

目的:探讨NLRP3抑制剂CY-09在血吸虫病肝纤维化中的作用。方法:采用BALB/c小鼠,构建CY-09干预的血吸虫病肝纤维化模型。通过HE、Masson和天狼星红染色观察小鼠肝脏组织中病理学变化,通过RT-qPCR和Western-blot等方法检测小鼠肝脏组织中NLRP3炎性小体活化和肝脏纤维化情况。Hyp试剂盒检测小鼠肝脏组织中羟脯氨酸含量,ELISA试剂盒检测小鼠血清中IL-1β含量。结果:HE染色显示,对照组小鼠肝小叶结构完整,肝细胞核大、圆,呈蓝色,细胞浆呈现粉色;模型组小鼠肝脏出现虫卵且肝小叶结构已经完全破坏,可见增多的炎性细胞和周围沉积的胶原纤维。天狼星红染色显示对照组肝组织被染成黄色或者淡黄色,而模型组小鼠肝组织中虫卵周围的大量胶原纤维被染成红色。与对照组小鼠相比较,模型组小鼠肝脏α-SMA、Collagen-Ⅰα1 mRNA表达增加(P<0.05),Hyp含量升高(P<0.05),NLRP3、pro-caspase-1、Caspase-1、pro-IL-1β和IL-1β蛋白表达增加(P<0.05)。与模型组小鼠相比,CY-09干预组小鼠肝组织Caspase-1和IL-1β蛋白表达显著降低。ELISA结果显示,与模型组小鼠相比,CY-09干预组小鼠血清中IL-1β含量显著降低(P<0.05)。与模型组小鼠比较,CY-09干预组小鼠的虫卵所致肝纤维化程度减轻。结论:CY-09可抑制血吸虫病肝纤维化小鼠模型中的NLRP3炎性小体活化,从而减轻肝纤维化。

Objective:To investigate the role of NLRP3 inhibitor CY-09 in hepatic fibrosis of schistosomiasis.Methods:A CY-09 intervention model of liver fibrosis in schistosomiasis was developed by using BALB/c mice.The pathological changes were observed in the liver tissue of mice by HE staining,Masson staining and Sirius red staining.The NLRP3 inflammasome activation and liver fibrosis were detected in the mice by RT-qPCR and Western-blot.The content of hydroxyproline in mouse liver tissue was detected by the Hyp kit and the content of IL-1βin mouse serum was determined by ELISA kit.Results:The HE staining showed that the liver lobule structure of the mice was structurally intact in the control group.The liver cell nuclei of the mice in this group is large and round,which is blue,and the cytoplasm is pink.The eggs were appeared in the liver of the mice,the liver lobule structure had been damaged,and inflammatory cells and collagen fibrosis could be seen in the model group.The Sirius red staining showed that the liver tissues were stained yellow or pale yellow in the control group,while a large amount of collagen fibers around the eggs were stained red in the liver of the mice in the model group.Compared with mice in the control group,the mRNA expression ofα-SMA and Collagen-Iα1 was increased in the liver of the mice in the model group(P<0.05),and the content of Hyp was higher than that in the control group(P<0.05).Compared with mice in the control group,the protein expression of NLRP3,pro-caspase-1,Caspase-1,pro-IL-1βand IL-1βwas increased in the liver of the mice in the model group(P<0.05).Compared with the model group,the protein expression of Caspase-1 and IL-1βwas significantly reduced in the liver of the mice in the CY-09 intervention group.The ELISA results showed that compared with the model group,the content of IL-1βwas significantly reduced in the serum of mice in the CY-09 intervention group(P<0.05).Compared with the model group,the area of liver fibrosis caused by the eggs was decreased in the CY-09 intervention group.Conclusion:CY-09 may inhibit NLRP3 inflammasome activation in the mice with a protective effect on liver fibrosis caused by schistosomiasis.