nephrin、podocin和podocalyxin mRNA在肾脏损伤患儿尿液中的表达情况分析
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摘要
目的探究足细胞相关分子在尿液中的表达在临床诊断患儿肾脏损伤中的研究价值及意义。方法选择2016年1-3月深圳市儿童医院肾脏免疫科住院患儿85例作为试验组,选择同期于该院门诊体检且尿常规结果均排除血尿及蛋白尿的18例健康儿童作为健康对照组,用反转录实时荧光定量PCR(RT-qPCR)法检测试验组和健康对照组尿沉渣中nephrin,podocin及podocalyxin(PCX)的表达。根据尿常规结果将试验组分为血尿组、蛋白尿组及混合组(血尿合并蛋白尿组),并观察试验组中血清胱抑素C(CysC)和血肌酐(Scr)水平与PCX的相关性。结果 (1)试验组nephrin,podocin及PCX表达阳性率显著高于健康对照组,差异有统计学意义(χ~2=13.849、6.271、9.650,P<0.05)。(2)混合组与蛋白尿组和血尿组间podocin阳性率比较,差异有统计学意义(χ~2=8.965、6.115,P<0.05)。(3)PCX阳性组血清CysC水平显著高于PCX阴性组,血清Scr在PCX阳性组与阴性组中水平差异无统计学意义(t=1.299,P>0.05)。结论肾脏受损时足细胞相关分子nephrin、podocin和PCX mRNA水平表达异常增加,这可能是血尿发生的分子机制,联合检测血CysC有助于早期发现肾损伤。
Objective To investigate the value and significance of podocyte related molecules in urine in clinical diagnosis of renal injury in children.Methods 85 childre hospitalized in Department of Nephrology and Immunology in Shenzhen Children′s Hospital from January to March in 2016 were selected as the experimental group,and 18 healthy children who underwent health assessment and did not have hematuria and proteinuria in routine urinalysis results were selected as the healthy control group.The expression levels of nephrin,podocin and podocalyxin(PCX)in experimental group and control group were detected by reverse transcription qPCR(RT-qPCR).According to the routine urinalysis,the experimental group was divided into hematuria group,proteinuria group and mixed group(hematuria and proteinuria group).The correlation between serum cystatin C(CysC)and serum creatinine(Scr)levels and PCX in the experimental group was also observed.Results(1)In urine,the expressions of nephrin,podocin and podocalyxin in experimental group were significantly higher than those of control group,and the differences were statistically significant(χ~2=13.849,6.271,9.650,P<0.05).(2)The expression of podocin in mixed group was significantly higher than that in hematuria group and proteinuria group,and the difference was statistically significant(χ~2=8.965,6.115,P<0.05).(3)The levels of CysC in the group which expressed podocalyxin was significantly higher than the group without podocalyxin expression.There was no significant difference in Scr between them(t=1.299,P>0.05).Conclusion When the kidney is injured,the expression of podocyte related molecules nephrin,podocin and PCX mRNA increases abnormally,this may be the molecular mechanism of hematuria too.Combined detection of serum CysC is beneficial for early kidney injury diagnosis.
Objective To investigate the value and significance of podocyte related molecules in urine in clinical diagnosis of renal injury in children.Methods 85 childre hospitalized in Department of Nephrology and Immunology in Shenzhen Children′s Hospital from January to March in 2016 were selected as the experimental group,and 18 healthy children who underwent health assessment and did not have hematuria and proteinuria in routine urinalysis results were selected as the healthy control group.The expression levels of nephrin,podocin and podocalyxin(PCX)in experimental group and control group were detected by reverse transcription qPCR(RT-qPCR).According to the routine urinalysis,the experimental group was divided into hematuria group,proteinuria group and mixed group(hematuria and proteinuria group).The correlation between serum cystatin C(CysC)and serum creatinine(Scr)levels and PCX in the experimental group was also observed.Results(1)In urine,the expressions of nephrin,podocin and podocalyxin in experimental group were significantly higher than those of control group,and the differences were statistically significant(χ~2=13.849,6.271,9.650,P<0.05).(2)The expression of podocin in mixed group was significantly higher than that in hematuria group and proteinuria group,and the difference was statistically significant(χ~2=8.965,6.115,P<0.05).(3)The levels of CysC in the group which expressed podocalyxin was significantly higher than the group without podocalyxin expression.There was no significant difference in Scr between them(t=1.299,P>0.05).Conclusion When the kidney is injured,the expression of podocyte related molecules nephrin,podocin and PCX mRNA increases abnormally,this may be the molecular mechanism of hematuria too.Combined detection of serum CysC is beneficial for early kidney injury diagnosis.
引文
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[6]FURUTA I,ZHAI T,ISHIKAWA S,et al.Association between nephrinuria,podocyturia,and proteinuria in women with preeclampsia[J].J Obstet Gynaecol Res,2017,43(1):34-41.
[7]ZHAI T,FURUTA I,AKAISHI R,et al.Alteration of podocyte phenotype in the urine of women with preeclampsia[J].Sci Rep,2016,6:24258.
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[9]林宜,朱斌,朱晓玲,等.肾小球疾病患者尿沉渣足细胞分子podocin和podocalyxin分析[J].中国中西医结合肾病杂志,2011,12(8):690-694.
[10]ARMELLONI S,CORBELLI A,GIARDINO L,et al.Podocytes:recent biomolecular developments[J].Biomol Concepts,2014,5(4):319-330.
[11]李莉,张碧丽.尿Podocalyxin鉴别儿童血尿来源的价值[J].实用中国儿科临床杂志,2011,26(5):357-359.
[12]SHOJI M,KOBAYASHI K,TAKEMOTO M,et al.Urinary podocalyxin levels were associated with urinary albumin levels among patients with diabetes[J].Biomarkers,2016,21(2):164-167.
[13]WICKMAN L,AFSHINNIA F,WANG S Q,et al.Urine podocyte mRNAs,proteinuria,and progression in human glomerular diseases[J].J Am Soc Nephrol,2013,24:2081-2095.
[14]DOS SANTOS M,BRINGHENTI R N,RODRIGUES P G,et al.Podocyte-associated mRNA profiles in kidney tissue and in urine of patients with active lupus nephritis[J].Int J Clin Exp Pathol,2015,8(5):4600-4613.
[15]罗琳.血清β2-微球蛋白、尿微量清蛋白、血清胱抑素C在高血压病早期肾损害诊断的意义[J].长江大学学报(自科版),2016,13(36):11-13.
[16]高娟梅.血清CysC和尿PCX在妊娠糖尿病早期肾损害诊断中的应用价值[J].中国优生与遗传杂志,2016,24(5):70-71.
[2]PATRAKKA J,TRYGGVASON K.Molecular make-up of the glomerular filtration barrier[J].Biochem Biophys Res Commun,2010,396(1):164-169.
[3]MA D,ZHONG S,LIU X,et al.CD3Dand PRKCQ work together to discriminate between B-cell and T-cell acute lymphoblastic leukemia[J].Comput Biol Med,2016,77:16-22.
[4]VOGELMANN S U,NELSON W J,MYERS B D,et al.Urinary excretion of viable Podocytes in health and renal disease[J].Am J Physiol Renal Physiol,2003,285(1):40-48.
[5]PEREIRA S V,DOS SANTOSM,RODRIGUES P G,et al.Increased urine podocyte-associated messenger rnas in severe obesity are evidence of podocyte injury[J].Obesity(Silver Spring),2015,23(8):1643-1649.
[6]FURUTA I,ZHAI T,ISHIKAWA S,et al.Association between nephrinuria,podocyturia,and proteinuria in women with preeclampsia[J].J Obstet Gynaecol Res,2017,43(1):34-41.
[7]ZHAI T,FURUTA I,AKAISHI R,et al.Alteration of podocyte phenotype in the urine of women with preeclampsia[J].Sci Rep,2016,6:24258.
[8]RODRIGUES P G,BRINGHENTI R N,NASCIMENTO J F,et al.Expression patterns of podocyte-associated mRNAs in patients with proliferative or non-proliferative glomerulopathies[J].Int J Clin ExP Pathol,2014,7(5):2185-2198.
[9]林宜,朱斌,朱晓玲,等.肾小球疾病患者尿沉渣足细胞分子podocin和podocalyxin分析[J].中国中西医结合肾病杂志,2011,12(8):690-694.
[10]ARMELLONI S,CORBELLI A,GIARDINO L,et al.Podocytes:recent biomolecular developments[J].Biomol Concepts,2014,5(4):319-330.
[11]李莉,张碧丽.尿Podocalyxin鉴别儿童血尿来源的价值[J].实用中国儿科临床杂志,2011,26(5):357-359.
[12]SHOJI M,KOBAYASHI K,TAKEMOTO M,et al.Urinary podocalyxin levels were associated with urinary albumin levels among patients with diabetes[J].Biomarkers,2016,21(2):164-167.
[13]WICKMAN L,AFSHINNIA F,WANG S Q,et al.Urine podocyte mRNAs,proteinuria,and progression in human glomerular diseases[J].J Am Soc Nephrol,2013,24:2081-2095.
[14]DOS SANTOS M,BRINGHENTI R N,RODRIGUES P G,et al.Podocyte-associated mRNA profiles in kidney tissue and in urine of patients with active lupus nephritis[J].Int J Clin Exp Pathol,2015,8(5):4600-4613.
[15]罗琳.血清β2-微球蛋白、尿微量清蛋白、血清胱抑素C在高血压病早期肾损害诊断的意义[J].长江大学学报(自科版),2016,13(36):11-13.
[16]高娟梅.血清CysC和尿PCX在妊娠糖尿病早期肾损害诊断中的应用价值[J].中国优生与遗传杂志,2016,24(5):70-71.