Role Of Acid Sensing Ion Channels In Renal Injury In Henoch Schonlein Purpura

Objective To observe the expression of acid-sensitive ion channels in renal tissues of children with Henoch-Schonlein purpura and the expression of ASICs in human renal tubular epithelial cells induced by extracellular acidification,and to explore their role in renal injury in children with Henoch-Schonlein purpura,and to further elucidate drug blocking ASICs may be a new target for the treatment of kidney damage in children with Henoch-Schonlein purpuraMethod1.A total of 13 children with Henoch-Schonlein purpura nephritis(HSPN)were enrolled in the pediatric and nephrology department of the First Affiliated Hospital of Anhui Medical University from January 2013 to January 2015.Among them,6 were male and 7 were female Aged 5 years 2 months to 13 years 4 months,after the kidney to collect their renal biopsy.7 cases of the control group,children with nephrotic syndrome,including 3 males and 4 females,the age of onset of 4 years 3 months to 14 years 1 month,to collect their normal renal biopsy.The expression of ASICs was detected in renal tissues of children with HSPN and simple nephrotic syndrome in the kit in accordance with the requirements of the immune group,while the quantitative expression of HSPN in 24 h urine protein and renal tissue ASICs correlation analysis.The expression of ASICs in HSPN was also analyzed according to the urine whether the?1-MG in the protein was abnormal was divided into normal group and abnormal group.According to whether the abnormality of ?2-MG was normal and abnormal group,the expression of ASICs in renal tubules of HSPN was observed.2.Human renal tubular epithelial cells(HKC)were used to study the effect of renal tubular epithelial cells at pH 7.4(control group),pH 7.0,pH 6.5,pH 6.0,pH5.5.The cells were cultured for 3 hours under the extracellular environment.After treated with the cells,the cells were cultured for 4 hours under normal culture medium.The expression of ASICs in renal tubular epithelial cells stimulated by different acids was determined by western blot.The renal tubular epithelial cells were grouped as follows:pH 7.4(Final concentration 100 ng / ml),pH6.5(control group),pH 6.5 + Amiloride(final concentration 100 umol / L),pH6.5 + Pc Tx-1(final concentration 100 ng / ml),in each group after administration of 30 min,adding pH6.5 cell culture media co culture 3h,change into normal training based pH7.4 cultured 4h,the cells were collected for detection of 30 min.Keratins was determined by Westernblot(K7,K18,K19)protein expression.Results1.Expression of ASICs in Renal Tissues of HSPN Children High ASICs expression was observed in renal tissue with HSPN patients,especially in renal tubular.2.Expression of ASICs in renal tubules of children with different proteinuria The quantitative expression of renal tubules in children with HSPN was positively correlated with 24 h urinary protein quantification(r = 0.644,P = 0.017).The expression of ASIC3 in ?1-MG abnormal group and ?2-MG abnormal group was significantly higher than that in normal group(P <0.01).The expression of ASIC2 a in ?1-MG abnormal group was higher than that in normal group(P <0.05).3.Expression of ASICs in renal tubular epithelial cells stimulated by different acid.The expression of ASIC1 a,ASIC2a and ASIC3 in renal tubular epithelial cells at pH7.0,pH6.5,pH6.0 and pH5.5 was significantly higher than that in pH7.4(control group),the difference was statistically significant.4 Keratins(K7,K18,K19)protein expression.The expression of Keratins(K7,K18 and K19)in pH6.5 group was significantly higher than that in group pH7.4(P<0.01).K18 in pH6.5 + Amiloride(final concentration 100umol/L)group and pH6.5 + Pc Tx-1(final concentration 100ng/ml)group compared with pH6.5(control group)reduced expression in renal tubular epithelial cells(P<0.05),K7,K19 in pH6.5 + Amiloride(final concentration 100umol/L)group and pH6.5 + Pc Tx-1(final concentration100ng/ml)group than pH6.5 group in renal tubular epithelial cell expression was significantly reduced(P<0.01).Conclusions:High ASICs expression was observed in renal tissue with HSPN patients,especially in renal tubular.ASICs may have a certain relationship with the production of proteinuria,especially microalbuminuria.Extracellular acid could induce the expression of ASICs in renal tubular epithelial cells and renal tubular injury,which was proved by increased Keratins K7?18?19.Blocking ASICs could reduce Keratins K7?18?19expression and protect the renal epithelial injury.