Dynamic changes of liver macrophage polarization induced by Clonorchis sinensis infection in mice
The study was to analyze the dynamic changes of liver macrophage polarization in mice during Clonorchis sinensis infection.In this study,HE staining was used to detect the changes of liver pathology and inflammatory infiltration at different stages of infection,and flow cytometry was used to detect the proportion of M1/M2 liver macrophages at different stages of infection.Subsequently,mRNA levels of M1-related markers iNOS,CD86 and IL-1β and M2 related markers Arg1,CD206 and Ym1 were detected by qRT-PCR at different stages of infection.The secretion levels of macrophage-related pro-inflammatory factors TNF-a and IL-1β,and anti-inflammatory factors TGF-β1 and IL-10 were detected by ELISA.Finally,to comprehensively analyze the dynamic changes of M1/M2 liver macrophage polarization in mice during Clonorchis sinensis infection.The results demonstrated that:1)Following infection with Clonorchis sinensis,the HE staining results of the mice liver exhibited a gradual transformation of cells from a normal morphology to fibrosis.Additionally,the inflammatory infiltration was more pronounced during the initial 1-2 weeks of infection,subsequently declining during the subsequent 6-10 weeks.2)The results of flow cytometry demonstrated a notable increase in the proportion of M1-type hepatic macrophages relative to M2-type hepatic macrophages during the initial 1-2 week period of infection(P＜0.01).Conversely,the proportion of M2-type hepatic macrophages exhibited a significant rise in comparison to M1-type hepatic macrophages during the subsequent 3-10 week phase of infection(P＜0.01).3)The results of qRT-PCR revealed a notable increase in the levels of M1-type markers,including iNOS,CD86,and IL-1β mRNA,during the initial 1-4 weeks of infection.This was followed by a decline and subsequent rise in these levels during the subsequent 6-10 weeks(P＜0.01).In contrast,the levels of M2-type markers,such as Arg1,CD206,and Ym1 mRNA,exhibited a gradual increase throughout the 1～10 weeks of infection.4)ELISA results indicated that the levels of pro-inflammatory cytokines TNF-a and IL-1β tended to increase significantly and then decrease at 1-4 weeks of infection,and increased significantly at 6～10 weeks of infection(P＜0.01).The levels of the anti-inflammatory cytokines TGF-β1 and IL-10 exhibited a gradual increase over the course of 1-10 weeks of infection.In conclusion,M1-type macrophages were found to be elevated and predominant during the early stages of infection,while M2-type macrophages were elevated and predominant during the middle and late stages of infection.Additionally,the polarization ratio of Mi-type and M2-type macrophages shifted during the egg-production stage.This study provides insights into the potential for better control of clonorchiasis by analyzing the dynamics of hepatic macrophage polarization.
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