The Effect Of Hydatid Cyst Fluid On The Viability Of Liver Cells And Its Mechanism

Objective:1.Detecting the expression of apoptosis,autophagy related markers and DDIT4 in the liver tissue of patients with hydatidosis alveolaris(HAE),to explore the role of apoptosis,autophagy and DDIT4 in hydatidosis alveolaris and the mechanism of their interaction.2.To explore the effect of echinococcus cyst fluid on the viability of hepatocytes in vitro,and to verify whether echinococcus cyst fluid can induce apoptosis.Methods: 1.Caspase 3,Bax,Bcl-2,lc3-I,LC3-II were detected by Immunohistochemistry,Q-PCR and Western blot,the expression of beclin-1,m TOR and ddit4,and the correlation between DDIT4 and apoptosis,autophagy related markers were analyzed.2.The proliferation ability and cell viability of mouse hepatocyte AML-12 cultured in vitro were detected by the intervention of Echinococcus derived cyst fluid,and the effect of Echinococcus derived cyst fluid on hepatocyte was evaluated by flow cytometry apoptosis detection.Results: 1.The results of immunohistochemistry,Q-PCR and Western blot showed that: caspase 3,Bax,lc3-I,LC3-II,The expression of beclin-1 and DDIT4 m RNA and protein in hae marginal zone and normal liver tissue were different,and the expression of these indexes in hae marginal zone was higher than that in normal liver tissue,while Bcl-2 and m TOR were higher The expression of m RNA and corresponding protein in liver tissue of hae marginal zone and normal liver tissue was lower than that in normal liver tissue(P < 0.05,with statistical significance);2.The results of in vitro experiments showed that Echinococcus fluid could inhibit the proliferation and cell viability of mouse hepatocytes,and the apoptotic rate of mouse hepatocytes after the treatment of Echinococcus fluid was higher than that of the control group,the optimal volume was 125 ul / 2.5ml medium,P < 0.05,which was statistically significant.Conclusion:Echinococcus cyst fluid may affect the proliferation of hepatocytes and the disappearance of hepatocytes around the focus by inducing apoptosis,autophagy and the expression of ddit4,which may be one of the pathogenesis of echinococcosis.