The relationship between the levels of renalase and changes in proteinuria, hypertension, renal function, renal tubular epithelial cell apoptosis and B-cell lymphoma-2 （Bcl-2） expression was investigated in patients...The relationship between the levels of renalase and changes in proteinuria, hypertension, renal function, renal tubular epithelial cell apoptosis and B-cell lymphoma-2 （Bcl-2） expression was investigated in patients （chronic nephritis, primary nephrotic syndrome or other kidney disease） that underwent renal biopsy. The study group comprised 72 patients undergoing renal biopsy. Patient profiles and renal function were collected. Concentrations of renalase and Bcl-2 were measured by immunohistochemistry. Tubular injury was detected by periodic acid Schiff staining （PAS） and renal tubular epithelial cell apoptosis was assessed by TUNEL assay. The expression of renalase was significantly lower in renal biopsy specimens than in normal kidney tissues. There was a positive linear relationship between renalase and some serum and cardiac indices; a negative correlation was found between age, eGFR, Ccr and 24-h urinary protein. Renal tubule injury index and tubular epithelial cell apoptosis index showed a negative linear correlation with renalase. The results showed that renalase probably increased the expression of Bel-2. By two independent samples t-test, renalase levels were significantly increased in the non-hypertension group than in the hypertension group. One-way ANOVA showed that renalase expression was higher in samples with Lee＇s grade Ⅲ than in those with Lee＇s grade V. The expression of renalase was significantly decreased in patients who underwent renal biopsy, and was also associated with blood and renal function. The research proved that renalase may reduce renal tubular injury and apoptosis of renal tubular epithelial cells through the mitochondrial apoptosis pathway, finally achieving the purpose of delaying the progress of renal failure.展开更多
AIM:To study the effect of renalase peptide,RP-220,on cell viability of human corneal epithelial cells after alkali insult.METHODS:A dose-response relationship between cell viability and exposure to NaOH solution were...AIM:To study the effect of renalase peptide,RP-220,on cell viability of human corneal epithelial cells after alkali insult.METHODS:A dose-response relationship between cell viability and exposure to NaOH solution were characterized using cultured human corneal epithelial cells.Viability of corneal epithelial cells was determined using commercially available MTT and CyQUANT?assays.RESULTS:At a concentration of 6 mmol/L,insult with NaOH leads to reduced corneal epithelial cell viability by approximately 30%.This reduced viability was prevented by treating the cells after initial insult with the 20-amino acid renalase derived peptide(RP-220).CONCLUSION:RP-220 has a pro-survival role for RP-220 following alkaline insult to corneal epithelial cells.展开更多
Introduction: Chronic kidney disease (CKD) is an important public health problem. Early detection and treatment is a key factor for prevention of its complications. Hypertensive nephrosclerosis is a subtype of CKD whi...Introduction: Chronic kidney disease (CKD) is an important public health problem. Early detection and treatment is a key factor for prevention of its complications. Hypertensive nephrosclerosis is a subtype of CKD which has a poor correlation between hypertension and development of nephropathy, implying role of genetic factors or epigenetic factors. The knowledge regarding genetic factors is limited. Renalase is a novel hormone with its gene on chromosome 10, which secretes flavin adenine dinucleotide dependent amine oxidase. Renalase metabolizes circulating catecholamines and modulates blood pressure and cardiac function. Recently, two single nucleotide polymorphisms of renalase gene rs2576178 GG and rs2296545 CC have been linked to essential hypertension. The SNPrs2296545 CC is also shown to be associated with cardiac hypertrophy, dysfunction and ischemia. The association of these two single nucleotide polymorphisms with hypertensive nephrosclerosis has not been investigated. Methods: We designed a case-control study to investigate whether the two known renalase gene polymorphisms rs2576178 and rs2296545 are associated with CKD particularly hypertensive nephrosclerosis. We genotyped these two polymorphisms in 287 subjects from North Indian population (106 CKD cases and 181 controls). Results: Comparison shows that subjects with hypertensive nephrosclerosis had higher frequencies of rs2296545 Callele than the healthy controls (0.63 versus 0.47, p 0.02). The odds ratio for rs2296545 CC genotype in hypertensive nephrosclerosis were 2.55 (95% CI, 1.03 to 6.42;p = 0.02) (CC versus GG) and 2.11(95% CI, 1.01 to 4.42;p = 0.03) (CC versus CG + GG) compared to controls. Conclusion: These findings may provide novel insight into the role of additional genomic regions as susceptibility gene in the pathophysiology of hypertensive nephrosclerosis. Further, to account for geoethnic variation, studies on heterogeneous populations involving a larger sample size are required. The correlation between this structural change and actual levels of the enzyme or the activity are required to strengthen this association as well as to be clinically applicable.展开更多
目的探讨血清Renalase(SRNLS)与慢性肾脏病(CKD)患者肾功能的相关性,明确其预测CKD患者肾功能的价值及影响SRNLS的危险因素。方法收集CKD 1期~5期非透析患者277例及健康对照者20例,检测血红蛋白(Hb)、血肌酐(Scr)、尿素氮(BUN)、血白蛋...目的探讨血清Renalase(SRNLS)与慢性肾脏病(CKD)患者肾功能的相关性,明确其预测CKD患者肾功能的价值及影响SRNLS的危险因素。方法收集CKD 1期~5期非透析患者277例及健康对照者20例,检测血红蛋白(Hb)、血肌酐(Scr)、尿素氮(BUN)、血白蛋白(Alb)、总胆固醇(TC)、甘油三酯(TG)及24 h尿蛋白定量(24 h Upro),计算估算的肾小球滤过率(eGFR),ELISA法检测SRNLS浓度。对CKD患者SRNLS分别与各临床参数做相关性分析;通过logistic回归分析明确影响CKD患者SRNLS的危险因素。结果与对照组相比,CKD患者血Scr、BUN、24 h Upro及SRNLS均升高,而Alb、Hb及eGFR均降低。CKD1期患者SRNLS与对照组无明显差异,但从CKD2期开始,患者SRNLS即开始升高,随着eGFR下降,SRNLS呈逐步升高的趋势。相关性分析发现,SRNLS与Scr(r=0.343,P<0.001)及24 h Upro(r=0.657,P=0.000)成正相关,与eGFR(r=-0.311,P=0.000)及Hb(r=-0.326,P=0.000)成负相关。logistic回归分析显示,糖尿病、Scr、BUN、Alb、eGFR、Hb及24h Upro均是影响CKD患者SRNLS浓度的危险因素(P<0.05)。结论CKD患者SRNLS浓度随肾功能恶化呈逐步升高趋势,是预测CKD患者肾功能的有效血清学标记物,而糖尿病、Scr、BUN、Alb、eGFR、Hb及24 h Upro是影响CKD患者SRNLS浓度的危险因素。展开更多
To obtain the recombination protein of renalase and prepare the monoclonal antibody, the human renalase gene was amplified from human kidney by specific primer and cloned the DNA fragments into the pET22b. After verif...To obtain the recombination protein of renalase and prepare the monoclonal antibody, the human renalase gene was amplified from human kidney by specific primer and cloned the DNA fragments into the pET22b. After verification of the positive clones, the gene was transformed to E. coli BL21 to express the protein with 6His on C terminal. The Balb/c mouse was immunized with the purified protein to prepare the monoclonal antibody by hybidoma technique. The renalase protein was reconstructed and 2 strains of the hybidoma which can stable secrete renalase were obtained. The monoclonal antibody can both react with the both recombinant and human serum renalase.展开更多
Heart rate variability(HRV) is a noninvasive technique for autonomic nerve activity assessment and an indirect index of adrenergic cardiovascular drive. Renalase is the only known enzyme degrading circulatory catechol...Heart rate variability(HRV) is a noninvasive technique for autonomic nerve activity assessment and an indirect index of adrenergic cardiovascular drive. Renalase is the only known enzyme degrading circulatory catecholamines, participating in regulations of the cardiovascular and sympathetic nervous systems. This study aims at the relation between serum renalase and HRV indexes in dialysis patients. There were 14 hemodialysis(HD)cases, 16 continuous ambulatory peritoneal dialysis(PD) cases, and 16 healthy volunteers enrolled in this study.All the subjects underwent 24 h Holter testing by using a dynamic electocardiogram monitor system to determine HRV parameters, and serum renalase levels were measured. The standard deviation of normal to normal R-R intervals(SDNN, a measure of beat-to-beat variability), root mean square successive difference(RMSSD), and the proportion derived by dividing NN50(the number of interval differences of successive normal to normal intervals greater than 50 ms) by the total number of NN intervals(pNN50) were determined to assess HRV. Both HD and PD patients presented increased maximum heart rate(Max-HR), minimum heart rate(Min-HR) and average heart rate(Ave-HR), and presented decreased SDNN, RMSSD and pNN50 relative to healthy controls. Serum levels of renalase were highly negatively correlated with Ave-HR(correlation coefficient r =-0.68, probability P < 0.01), while positively correlated with SDNN(r = 0.64, P < 0.01) in both the HD and PD groups. Serum level of renalase was positively correlated with 24 h urine volume(r = 0.73, P < 0.01) in PD patients. Renalase might be reciprocally related to HRV and inversely related to Ave-HR in dialysis patients, which will help to probe into renalase's physiology and may pave the way to explore renalase replacement therapy in patients with end stage renal disease(ESRD).展开更多
Renalase is a secreted amine oxidase that metabolizes catecholamines.It has been proposed to modulate blood pressure and heart rate and its downregulation might result in hypertension.Despite its potential relevance f...Renalase is a secreted amine oxidase that metabolizes catecholamines.It has been proposed to modulate blood pressure and heart rate and its downregulation might result in hypertension.Despite its potential relevance for human health,the biochemical characterization of renalase is still scarce.The aim of this study is to synthesize the human renalase eukaryotic protein by genetic engineering.The human renalase gene was amplified by polymerase chian reaction (PCR).After digestion by BamH I and Xho I enzymes,the DNA fragments were cloned into the transfer vector,pFastBacHTb-Fc,to generate the pFastBacHTb-renalase expression vector.The ligation products were transformed into E.coli DH10Bac to obtain recombinant transposon rBacmid-renalase.The recombinant transposon was further transferred into insect high-V cells,and the recombinant human renalase eukaryotic protein was expressed successfully.展开更多
文摘The relationship between the levels of renalase and changes in proteinuria, hypertension, renal function, renal tubular epithelial cell apoptosis and B-cell lymphoma-2 （Bcl-2） expression was investigated in patients （chronic nephritis, primary nephrotic syndrome or other kidney disease） that underwent renal biopsy. The study group comprised 72 patients undergoing renal biopsy. Patient profiles and renal function were collected. Concentrations of renalase and Bcl-2 were measured by immunohistochemistry. Tubular injury was detected by periodic acid Schiff staining （PAS） and renal tubular epithelial cell apoptosis was assessed by TUNEL assay. The expression of renalase was significantly lower in renal biopsy specimens than in normal kidney tissues. There was a positive linear relationship between renalase and some serum and cardiac indices; a negative correlation was found between age, eGFR, Ccr and 24-h urinary protein. Renal tubule injury index and tubular epithelial cell apoptosis index showed a negative linear correlation with renalase. The results showed that renalase probably increased the expression of Bel-2. By two independent samples t-test, renalase levels were significantly increased in the non-hypertension group than in the hypertension group. One-way ANOVA showed that renalase expression was higher in samples with Lee＇s grade Ⅲ than in those with Lee＇s grade V. The expression of renalase was significantly decreased in patients who underwent renal biopsy, and was also associated with blood and renal function. The research proved that renalase may reduce renal tubular injury and apoptosis of renal tubular epithelial cells through the mitochondrial apoptosis pathway, finally achieving the purpose of delaying the progress of renal failure.
基金Supported by the resources of the Central Texas Veterans Health Care System (Temple, TX)the Central Texas Veterans Health Care System Research Service
文摘AIM:To study the effect of renalase peptide,RP-220,on cell viability of human corneal epithelial cells after alkali insult.METHODS:A dose-response relationship between cell viability and exposure to NaOH solution were characterized using cultured human corneal epithelial cells.Viability of corneal epithelial cells was determined using commercially available MTT and CyQUANT?assays.RESULTS:At a concentration of 6 mmol/L,insult with NaOH leads to reduced corneal epithelial cell viability by approximately 30%.This reduced viability was prevented by treating the cells after initial insult with the 20-amino acid renalase derived peptide(RP-220).CONCLUSION:RP-220 has a pro-survival role for RP-220 following alkaline insult to corneal epithelial cells.
文摘Introduction: Chronic kidney disease (CKD) is an important public health problem. Early detection and treatment is a key factor for prevention of its complications. Hypertensive nephrosclerosis is a subtype of CKD which has a poor correlation between hypertension and development of nephropathy, implying role of genetic factors or epigenetic factors. The knowledge regarding genetic factors is limited. Renalase is a novel hormone with its gene on chromosome 10, which secretes flavin adenine dinucleotide dependent amine oxidase. Renalase metabolizes circulating catecholamines and modulates blood pressure and cardiac function. Recently, two single nucleotide polymorphisms of renalase gene rs2576178 GG and rs2296545 CC have been linked to essential hypertension. The SNPrs2296545 CC is also shown to be associated with cardiac hypertrophy, dysfunction and ischemia. The association of these two single nucleotide polymorphisms with hypertensive nephrosclerosis has not been investigated. Methods: We designed a case-control study to investigate whether the two known renalase gene polymorphisms rs2576178 and rs2296545 are associated with CKD particularly hypertensive nephrosclerosis. We genotyped these two polymorphisms in 287 subjects from North Indian population (106 CKD cases and 181 controls). Results: Comparison shows that subjects with hypertensive nephrosclerosis had higher frequencies of rs2296545 Callele than the healthy controls (0.63 versus 0.47, p 0.02). The odds ratio for rs2296545 CC genotype in hypertensive nephrosclerosis were 2.55 (95% CI, 1.03 to 6.42;p = 0.02) (CC versus GG) and 2.11(95% CI, 1.01 to 4.42;p = 0.03) (CC versus CG + GG) compared to controls. Conclusion: These findings may provide novel insight into the role of additional genomic regions as susceptibility gene in the pathophysiology of hypertensive nephrosclerosis. Further, to account for geoethnic variation, studies on heterogeneous populations involving a larger sample size are required. The correlation between this structural change and actual levels of the enzyme or the activity are required to strengthen this association as well as to be clinically applicable.
文摘目的探讨血清Renalase(SRNLS)与慢性肾脏病(CKD)患者肾功能的相关性,明确其预测CKD患者肾功能的价值及影响SRNLS的危险因素。方法收集CKD 1期~5期非透析患者277例及健康对照者20例,检测血红蛋白(Hb)、血肌酐(Scr)、尿素氮(BUN)、血白蛋白(Alb)、总胆固醇(TC)、甘油三酯(TG)及24 h尿蛋白定量(24 h Upro),计算估算的肾小球滤过率(eGFR),ELISA法检测SRNLS浓度。对CKD患者SRNLS分别与各临床参数做相关性分析;通过logistic回归分析明确影响CKD患者SRNLS的危险因素。结果与对照组相比,CKD患者血Scr、BUN、24 h Upro及SRNLS均升高,而Alb、Hb及eGFR均降低。CKD1期患者SRNLS与对照组无明显差异,但从CKD2期开始,患者SRNLS即开始升高,随着eGFR下降,SRNLS呈逐步升高的趋势。相关性分析发现,SRNLS与Scr(r=0.343,P<0.001)及24 h Upro(r=0.657,P=0.000)成正相关,与eGFR(r=-0.311,P=0.000)及Hb(r=-0.326,P=0.000)成负相关。logistic回归分析显示,糖尿病、Scr、BUN、Alb、eGFR、Hb及24h Upro均是影响CKD患者SRNLS浓度的危险因素(P<0.05)。结论CKD患者SRNLS浓度随肾功能恶化呈逐步升高趋势,是预测CKD患者肾功能的有效血清学标记物,而糖尿病、Scr、BUN、Alb、eGFR、Hb及24 h Upro是影响CKD患者SRNLS浓度的危险因素。
基金the Scientific Research Project of Shanghai Municipal Health Bureau (2008Y034)the Natural Scientific Research Project of Shanghai(05ZR14086)
文摘To obtain the recombination protein of renalase and prepare the monoclonal antibody, the human renalase gene was amplified from human kidney by specific primer and cloned the DNA fragments into the pET22b. After verification of the positive clones, the gene was transformed to E. coli BL21 to express the protein with 6His on C terminal. The Balb/c mouse was immunized with the purified protein to prepare the monoclonal antibody by hybidoma technique. The renalase protein was reconstructed and 2 strains of the hybidoma which can stable secrete renalase were obtained. The monoclonal antibody can both react with the both recombinant and human serum renalase.
基金the National Natural Science Foundation of China(Nos.81100528 and 81270824)the New 100 Talent Plan of Shanghai Jiaotong University School of Medicinethe Shanghai Talents Development Fund
文摘Heart rate variability(HRV) is a noninvasive technique for autonomic nerve activity assessment and an indirect index of adrenergic cardiovascular drive. Renalase is the only known enzyme degrading circulatory catecholamines, participating in regulations of the cardiovascular and sympathetic nervous systems. This study aims at the relation between serum renalase and HRV indexes in dialysis patients. There were 14 hemodialysis(HD)cases, 16 continuous ambulatory peritoneal dialysis(PD) cases, and 16 healthy volunteers enrolled in this study.All the subjects underwent 24 h Holter testing by using a dynamic electocardiogram monitor system to determine HRV parameters, and serum renalase levels were measured. The standard deviation of normal to normal R-R intervals(SDNN, a measure of beat-to-beat variability), root mean square successive difference(RMSSD), and the proportion derived by dividing NN50(the number of interval differences of successive normal to normal intervals greater than 50 ms) by the total number of NN intervals(pNN50) were determined to assess HRV. Both HD and PD patients presented increased maximum heart rate(Max-HR), minimum heart rate(Min-HR) and average heart rate(Ave-HR), and presented decreased SDNN, RMSSD and pNN50 relative to healthy controls. Serum levels of renalase were highly negatively correlated with Ave-HR(correlation coefficient r =-0.68, probability P < 0.01), while positively correlated with SDNN(r = 0.64, P < 0.01) in both the HD and PD groups. Serum level of renalase was positively correlated with 24 h urine volume(r = 0.73, P < 0.01) in PD patients. Renalase might be reciprocally related to HRV and inversely related to Ave-HR in dialysis patients, which will help to probe into renalase's physiology and may pave the way to explore renalase replacement therapy in patients with end stage renal disease(ESRD).
基金the Natural Scientific Research Project of Shanghai (No.05ZR14086)the Scientific Research Project of Shanghai Municipal Health Bureau (No.2008Y034)
文摘Renalase is a secreted amine oxidase that metabolizes catecholamines.It has been proposed to modulate blood pressure and heart rate and its downregulation might result in hypertension.Despite its potential relevance for human health,the biochemical characterization of renalase is still scarce.The aim of this study is to synthesize the human renalase eukaryotic protein by genetic engineering.The human renalase gene was amplified by polymerase chian reaction (PCR).After digestion by BamH I and Xho I enzymes,the DNA fragments were cloned into the transfer vector,pFastBacHTb-Fc,to generate the pFastBacHTb-renalase expression vector.The ligation products were transformed into E.coli DH10Bac to obtain recombinant transposon rBacmid-renalase.The recombinant transposon was further transferred into insect high-V cells,and the recombinant human renalase eukaryotic protein was expressed successfully.
