Molecular Biological Mechanism Of Alveolar Echinococoosis Regulating EMT Of Hepatocytes Through Macrophage Polarization

Objective: 1)to examine the changes of various cytokines in the supernatant after coculture between protoscoliosis and macrophages(RAW264.7)to infer the type of macrophage polarization,2)to explore the effects of the supernatant of protoscoliosis coculture with macrophages on EMT in liver cells(AML12),and 3)to detect the alterations of EMT markers in marginal zone tissues and distant lesional tissues of patients with hepatic alveolar echinococcosis,initially to explore the effects of alveolar echinococcosis on liver injury and repair of mechanisms.Methods: 1)to measure the size of the length and area of macrophages cocultured with protoscoliosis,2)to detect the changes of various cytokines in the supernatant after protoscoliosis cocultured with macrophages using cytokine array,and 3)Co-culture the supernatant of the protosphagoid and macrophages with hepatocytes,and use real-time quantitative PCR(q PCR)and Western blot(Western Blot,WB)to detect changes in EMT markers(E-cadherin,N-cadherin,snail,vimentin,ZO-1,fibronectin),and 4)the differences of EMT markers between marginal zone tissue of people with hepatic alveolar echinococcosis and tissue away from the lesion were detected by q PCR,WB,immunohistochemistry(IHC),immunofluorescence(IF).Results: 1)the length and area of macrophages cocultured with macrophages were significantly larger(P < 0.05)than those cultured with macrophages only,and 2)Compared with simple macrophage culture,Eotaxin-2,M-CSF,s TNF RⅡ,and IL-13 cytokines in the supernatant after the co-culture of the protocrophage and macrophages were significantly increased,while CCL5 was significantly down-regulated(P < 0.05);3)after incubation of supernatants from the coculture of protoscoliosis and macrophages with hepatocytes,E-cadherin and ZO-1 were downregulated(P < 0.05),while vimentin and fibronectin were significantly increased(P < 0.05),and N-cadherin was unchanged(P > 0.05);4)Compared with tissues far away from the lesion,the expressions of E-Cadherin and ZO-1 in the marginal zone tissues of patients with alveolar hydatid were significantly reduced,while the expressions of Vimentin and Snail were increased(P <0.05),while Fibronectin and N-Cadherin did not change significantly.Conclusions: Protocrophage may regulate the transformation of hepatocytes from epithelial to mesenchymal through macrophage polarization.