Effects of BCG-infected macrophages on renal tubular epithelial cell injury and repair
Objective:To investigate effect of Mycobacterium tuberculosis BCG-infected macrophages on damage and repair of renal tubular epithelial cells during development of renal tuberculosis.Methods:A co-culture model of BCG-infected M0 macrophages(upper chamber)and HK-2 cells(lower chamber)was established by Transwell,and THP-1 human monocyte macrophages were induced by 100 ng/ml phorbol ester(PMA)24 h to become M0 macrophages,and BCG infection cell model was established.Total cell protein was collected at 12 h and 24 h of infection,respectively.Western blot was used to detect expressions of M1 macrophage marker CD86 and M2 macrophage marker CD206 protein.M1 macrophage polarization marker cytokines IL-6 and TNF-α and M2 macrophage polarization marker cytokine TGF-β expressions in cell culture supernatant were detected by ELISA;experiment was divided into HK-2 group,BCG+HK-2 group,BCG+M0+HK-2 group and M0+HK-2 group,CCK-8 was used to detect viability of HK-2 cells in each group,and Hoechst test was used to detect HK-2 cells apoptosis in each group.Epithelial cell marker E-cadhren and fibroblast markerα-SMA expressions in HK-2 cells of each group were detected by Western blot.Results:After BCG infection of M0 macrophages,M1 macrophage viability was higher than 24 h at 12 h(P<0.05),and M2 macrophage was higher than 12 h at 24 h(P<0.05).After two cells co-culture,HK-2 cell viability was higher than 12 h at 24 h(P<0.001),apoptosis level was higher than 24 h at 12 h,epithelial cell marker protein E-cadherin protein level was higher than 12 h at 24 h(P<0.001),fibroblast level of cell marker protein α-SMA protein at 12 h was higher than that at 24 h(P<0.01).Conclusion:During development of renal tuberculosis,early BCG-infected macrophages may promote inflammatory injury of renal tubular epithelial cells through M1-type polarization;with prolongation of infec-tion time,they may repair renal tubular epithelial cells through M2-type polarization and plays an important protective role.
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