| IntroductionDiabetic nephropathy(DN) is the furthest chronic syndrome among diabetes,and it is leading cause of end stage renal disease in the world. The development of strategies for treatment of diabetic nephropathy is a urgent issue,however,a clear cut strategy has not been established. Various factors are known to be involved in the progression of diabetic nephropathy,including hyperglycemia,hypertension,dyslipidemia,production of inflammatory cytokines and oxidative stress. Recent studies indicate that diabetic nephropathy is related to glomerular extracellular matrix(ECM) accumulation that leads to glomerulosclerosis. The mesangial cell plays a key role in the development of glomerulosclerosis in DN.The several recent studies have focuses attention on the role of hyperglycemia. Advanced glycosylation endproducts(AGEs) have been demonstrated to play a role in the development of DN. High ambient glucose level transforms intracellular pathways promoting stable phenotypic changes in the glomemlus such as mesangial cell hypertrophy,podocyte,apoptosis,and matrix expansion. This effect is mediated through AGEs and is TGF-βand CTGF-dependent. There is abundant evidence in the literature implicating TGF-βas the major fibrogenic growth factor in the pathogenesis of glomerulosclerosis and interstitial fibrosis. Its role in the development of DN has been established.Connective tissue growth factor(CTGF) as downstream media to TGF-βis another prosclerotic growth factor implicated in the pathogenesis of renal fibrosis in DN.CTGF belongs to CCN family that includes CTGF,CYR61 and NOV and is one of the immediate early gens.To date ,CTGF has been implicated in numerous physiological and pathological processes,such as cell proliferation,migration,extracellular matrix(ECM) production,cell attachment,cell survival and apoptosis.Base on the role of increasing expression of ECM,this study aimed to observe the effects of antisense CTGF oligodexynucleotide (CTGF ASODN) on the expression of extracellular matrix in cultured human glomerular mesangial cells(HGMC) under high glucose,and to investigate the effect of CTGF ASODN in the forepart treatment to DN.Materials and Methods1,Materialsprimary normal adult human mesangial were purchased from Sciencell Company(US),Lipofectamine 2000 were purchased from Invitrogen Life Technologies(US),CTGF ASODN(5'-CGC GAC GGA GGC GAG CAT-3') and SODN (5'-ATG CTC GCC TCC GTC GCG-3') were composed by Shanghai Sangon Biological Enginology&Serricos Co.Ltd. Human FN and LN ELISA kit were purchased from Shanghai Senxiong Company.2,MethodsHGMCs were cultured in vitro,and used passage 5-8 to experiment. HGMCs were divided into six groups:1. normal control group(glucose 5.0mmol/l), 2.high glucose group(glucose 30mmol/l), 3.high glucose+mellifluence(25mmol/l),4.high glucose+ CTGF ASODN,5.highglucose+CTGF SODN,6.high glucose+Lipofectamine.We transfected CTGF ODN with lipofectamine 2000.To collect HGMCs lysates in different times(0h,18h,24h,48h,72h)The secretion of fibronectin and laminin by HGMCs in supernatants of culture media was detected with ELISA.Results1. High glucose midia significantly increased the syntheses and secretion of fibronectin and laminin in cultured HGMCs in a time-dependent manner,however, mellifluence had no effects.2. Transfection of HGMCs with CTGF ASODN decreased the secretion of fibronectin and laminin in supernatants of HGMC lysates and culture media,but transfection of HGMCs with CTGF SODN and lipofectamine had no effects.ConclusionsHigh glucose promotes the expression of HGMCs extracellular matrix,Transfection of HGMCs with CTGF ASODN can decrease the role of high glucose,and attenuate the expression of extracellular matix in supernatants of HGMCs lysates and culture media. Modulation of CTGF expression in HGMCs may work as effective way to mitigate the progression of diabetic nephropathy.
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