The research of the protective effect of combined treatment of apigenin and ischemic post-conditioning on renal ische-mia/reperfusion injury use NRK-52E cell

LIU Yang; LIU Xiuheng; WANG Lei; DU Yang; WANG Zhishun

doi:10.3969/j.issn.1671-6450.2017.04.019

Abstract：Objective To investigate the effects of combined treatment of apigenin and ischemic post-conditioning on the expression of TLR4/NF-κB, using the kidney cortex proximal tubule epithelial NRK-52E cells in vitro ischemia/reperfu-sion model .Methods From July 2016 to November 2016 at the Wuhan University Hubei key laboratory of Digestive System Disease carried on this study .Establishing renal ischemia/reperfusion injury cells model .The kidney cortex proximal tubule epithelial NRK-52E cells were randomly divided into 5 groups:Normal group, I/R group, Api group, IPO group, Api+IPO group.CCK8 and flow cytometry were used to accessing apoptosis .The levels of SOD and MDA were evaluated by kits .The expression of TLR4, NF-κB, TNF-αand IL-1βwere performed by Real-time PCR and Western blot.Results Compared with the normal group, apoptosis rate was aggravated in I/R group ( q =15.33, P <0.01), and this rate was significantly decreased in Api group , IPO group and Api +IPO group ( q =7.46, q =8.92, q =14.06, P =0.00, P =0.00, P =0.00).Api and IPO could increase the level of SOD ( q =-3.49, q =-2.71, q =-4.97, P =0.02, P =0.04, P =0.00) and reduce the level of MDA induced by IRI ( q =2.95, q =2.29, q =4.07, P =0.03, P =0.04, P =0.01).Re-al-time PCR and Western blot showed that TLR 4, NF-κB, TNF-αand IL-1βexpression increased significantly in IRI group (q =-7.74, q =-9.52, q =-10.88, q =-10.75, P =0.00, P =0.00, P =0.00, P =0.00).And their expression decreased significantly in Api group , IPO group and Api+IPO group ( q =2.99, q =3.71, q =3.98, q =4.52;q =2.57, q =3.03, q =3.50, q =3.37;q =4.47, q =5.03, q =5.87, q =5.56, P =0.03, P =0.01, P =0.01, P =0.02;P =0.04, P =0.03, P =0.01, P =0.02;P =0.00, P =0.00, P =0.00, P =0.00).Conclusion In vitro the combined treatment of apigenin and ischemic post-conditioning significantly weakened the injury by ischemia /reperfusion and the mecha-nism may be through inhibiting TLR 4/NF-κB pathway to exert protective effects .