Renal protective effect of lentivirus-mediated adiponectin gene transfection on streptozocininduced early diabetic nephropathy mice / 中华肾脏病杂志

ObjectiveTo investigate the renal protective effect of recombinant lentivirus encoding adiponectin gene on streptozocin-induced early diabetic nephropathy(DN) mice,and to explore its potential mechanism.Methods Forty C57BL/6 mice were randomly divided into normal control group(NC group,n=10),diabetic nephropathy group(DN group,n=10),LentiIRES-EGFP treatment group(DL group,n=10) and Lenti-Acdc-IRES-EGFP treatment group (DA group,n=10).After 8 weeks of recombinant lentivirus injection,kidney to body weight ratio (KW/BW),mean glomerular volume(MGV),fractional mesangial area(FMA),24 h urinary protein excretion(UTP),Scr,BUN,serum albumin and adiponectin were measured.Renal pathological changes were evaluated by electron microscopy.Proliferation of glomendar and tubulointerstitial cells was assessed by immunohistochemistry using PCNA antibody.The phosphorylation of AMP-activated protein kinase(AMPK) and mammalian target of rapamycin protein(mTOR) were detected by Western blotting.Results Adiponectin was successfully over-expressed in STZ-induced DN mice after lentivirus injection.KW/BW,MGV,FMA and UTP were significantly decreased in DA group as compared to DN group and DL group(P＜0.05),but were increased as compared to NC group(P＜0.05).DA group animals had significantly fewer PCNA-positive cells than DN group and DL group(P＜0.01).DA group mice had higher p-AMPK level and lower p-mTOR level as compared to DN group and DL group(P＜0.01).Conclusion Over-expression of adiponectin has beneficial effect on early DN and attenuates aberrant proliferation of renal cells via AMPKmTOR pathway.

Effects of recombinant lentivirus encoding human apM1 gene on platelet-derived growth factor inducedmesangial cell proliferation / 中华内分泌代谢杂志

Objective To evaluate the effects of recombinant lentivirus encoding human apM1 gene ( LentiapM1-EGFP) on platelet-derived growth factor (PDGF) induced human mesangial cell (HMC) proliferation and intracellular AMP-activated protein kinase(AMPK) signaling pathway.Methods Protein expression of apM1 in cell culture supernatant of HMC transfected with Lenti-apM1-EGFP was detected by ELISA.The effect of human adiponectin on cell proliferation and cell cycle was assessed by [ 3 H ] thymidine incorporation assay and Flow cytometry respectively.The phosphorylation of AMPK was detected by Western blotting.Results Lenti-apM1-EGFP had no significant toxicity on HMC.The 50 multiplicity of infection (MOI) of the Lenti-apM1-EGFP efficiently infected HMC,and made it stable expression of adiponectin protein ( 149.6 ± 12.8 ) μg/L.PDGF-induced HMCs proliferation was significantly inhibited by adiponectin.When co-treatment with compound C,an AMPK inhibitor,the inhibitory effort was reversed.The phosphorylation level of AMPK was increased in HMC transfected Lenti-apM1-EGFP compared to that of control.Conclusions Adiponectin antagonizes stimulatory effect of platelet-derived growth factor on mesangial cell proliferation by AMPK signaling.