Mir - 29 B Purpura Nephritis In Renal Tissue Of The Expression And Function Research

Part Ⅰ MiR-29b expression in the renal tissue of HenochSch nlein purpura nephritisObjective To detect miR-29b expression in the renal tissue of Henoch Sch nleinpurpura nephritis (HSPN), and to explore its relationship with pathological damage ofHSPN.Methods35HSPN renal biopsy children were chosen as subjects, and pathologicalchanges were observed with light and electron microscopy. Children were dividedinto two groups according to crescent formation, glomerular sclerosis, interstitialfibrosis, thrombosis formation and capillary loops necrosis. Real-time PCR was usedto quantify miR-29b expression in renal biopsy tissue, and initial laboratoryparameters after hospitalized were collected. Data analysis of miR-29b expressionand laboratory parameters was performed using the SPSS17.0. The prediction modelwas constructed via logistic regression analysis combining miR-29b and laboratoryparameters, and we the analyzed their relationship with crescent formatin of HSPN.Results (1) Comparing with group of no crescent, miR-29b expression was increasedin crescent group (P<0.05), and it had no change in other pathological change groups.(2) Among the laboratory parameters of the two subgroups, the levels of platelet,neutrophilic granulocyte percentage, eosinophils percentage, urine white cell counts,urine red cell counts,24h urinary protein after weight adjusted, serum cholesterol andIgG have significant statistic differences (P<0.05).(3) Logistic regression multivariable analysis showed that three parameters including miR-29b (OR=2.166,P=0.041),24hUPro/kg (OR=5.296, P=0.042), URBC (OR=6.016, P=0.033) furtherentered the logistic regression model (The logistic equation: LogP=-4.633+0.773miR-29b+1.66724h UPro/kg+1.794URBC) to predict the crescent formationindependently.Conclusions miR-29b expression increases in the renal tissue of HSPN with crescentformation. miR-29b,24hUPro/kg and URBC were the independent risk factors ofcrescent formation. This prompted clinicians should pay close attention to thosepatients with the risk factors and give them active treatment for slowing or evenpreventing the formation of glomerular crescents, and finally minimized the damagesof kidney. Part Ⅱ MiR-29b mediates AngⅡ-induced mesangialcell activationObjective To explore the effect of miR-29b in AngⅡ-induced glomerular mesangialcells activation (proliferation and expression of inflammatory mediators).Methods To evaluate effect of AngⅡon human glomerular mesangial cells (HGMCs),HGMCs were cultured in vitro and treated with AngⅡat conceration of100nmol/L.LV-hsa-miR-29b-1was stably transfected into HGMCs for miR-29b overexpression,and LV-hsa-miR-29b-3p-inhibition was stably transfected into HGMCs for inhibitingmiR-29b expression before application of AngⅡstimulation. The expressions of miR-29b, ICAM-1, IL-1β, IL-6, IL-8, TNFAIP3, NKRF, CyclinA2and CyclinD1were detected by real-time PCR and western blot. IL-1β and IL-8were alsodetermined by ELISA. Cell cycle was determined by flow cytometry. Cellproliferation was detected by CCK-8.Results (1) AngⅡtreatment increased the expressions of miR-29b, inflammatorymediators such as ICAM-1, IL-1β, IL-6, IL-8and cell cycle proteins CyclinA2,CyclinD1, and increasing of S+G2/M phase of the cell cycle (P<0.05).(2) miR-29boverexpression induced the release of ICAM-1, IL-1β, IL-6,IL-8, increasing ofCyclinA2、CyclinD1, and cell proliferation, however, it also could inhibit theexpression of TNFAIP3and NKRF (P<0.05).(3) miR-29b interference protectedagainst AngⅡ-induced release of inflammatory mediators, increasing of cell cycleproteins and cell proliferation. Moreover, miR-29b interference could increase theexpressions of TNFAIP3and NKRF (P<0.05).Conclusions miR-29b induced release of inflammatory mediators via inhibiting theexpression of TNFAIP3and NKRF, and it also promoted cell proliferation viainducing the cell cycle proteins producing.