The effects of M2-type macrophages on renal tubular epithelial cells injury exposed to calcium oxalate crystal

LIU Quan; GUAN Xiao-feng; TAO Zhi-wei; WANG Xiang; WU Ji-hua; LIU Yun-long; HE Zi-qi; DENG Yao-liang

doi:10.3969/j.issn.1001-9448.2018.03.002

Abstract：Objective To investigate the effects of M2-type macrophages on renal tubular epithelial cells ( HK-2) injury exposed to calcium oxalate ( CaOx) crystal. Methods THP-1 cells were induced into M2-type macrophages by Phorbol-12-myristate-13-acetate ( PMA) , IL-4 and IL-13. Their phenotypic markers were identified by RT-PCR, Western Blot and ELISA. HK-2 cells were incubated with CaOx at a concentration of 0. 5 mg /mL. The co-cultured model of M2-type macrophages ( upper chamber) and HK-2 cells ( lower chamber) were constructed by Transwell. The experiment was divided into HK-2 cells group, HK-2 cells + M2-type macrophages co-cultured group, HK-2 cells + CaOx group, and HK-2 cells + CaOx + M2-type macrophages co-cultured group. The viability of HK-2 cells was detected by CCK-8. The activity of lactate dehydrogenase ( LDH) in the supernatant of HK-2 cells was detected by microplate reader. DAPI was used for the apoptosis staining and Western blot was used for detecting the expression of Bcl-2 in HK-2 cells. The effects of M2 macrophages on the migration and repairmen of HK-2 cells were observed by the scarification test. Results THP-1 cells were successfully induced into M2-type macrophages. The CCK-8 test showed that M2-type macrophages significantly increased the viability of CaOx on HK-2 cells and M2-type macrophages proliferated on HK-2 cells ( P＜ 0. 05) . The LDH test showed that M2 macrophages significantly reduced the LDH of HK-2 cells exposed to CaOx ( P＜ 0. 05) . The DAPI test and the Bcl-2 expression showed that M2 macrophages inhibited the HK-2 cells from apoptosis exposed to CaOx. The scarification experiment showed that M2-type macrophages had a role of promoting the migration and repairmen to HK-2 cells. Conclusion M2-type macrophages have a certain inhibitory effect on HK-2 cells injury exposed to CaOx and can also promote the proliferation, migration and repairmen of HK-2 cells.