尿酸、尿素对HK-2细胞凋亡、纤维化相关因子表达的影响及前列地尔的干预研究
|
摘要
第一部分尿酸对人肾小管上皮细胞的凋亡、纤维化相关因子表达的影响及PGE1干预的体外研究
目的：研究尿酸对人肾小管上皮细胞凋亡、纤维化相关因子表达的影响以及PGE1的干预作用。
方法：①.培养HK-2细胞；②.不同浓度的尿酸刺激HK-2细胞24h、48h、72h,用MTT比色法检测细胞活性抑制率；不同浓度尿酸培养HK-2细胞24h,用Annexin V. FITC/PI流式细胞检测凋亡及检测NAG酶的释放量,来观察尿酸对HK-2细胞的毒性作用；通过Western Blot和Real-Time PCR检测TGF-β1、CTGF、FN的表达,观察尿酸对HK-2细胞的促纤维化作用；并加入PGE1了解其对HK-2细胞的保护作用。
结果：①.HK-2细胞加入不同浓度的尿酸分别作用24h、48h、72h后,与对照组比较,细胞活性抑制率明显升高,并呈时间剂量依赖性。尿酸作用HK-2细胞24h后,与对照组比较,细胞凋亡率呈剂量依赖性增加至56.06%±4.937%,NAG酶释放量也呈剂量依赖性增加至15.23±0.617。800umol/L尿酸组加入PGE1后,与800umol/L尿酸组比较,细胞活性抑制率下降明显,细胞凋亡率也明显降低至41.61%±4.149%,NAG酶释放量明显减少至11.24±0.468。②.用不同浓度的尿酸作用HK-2细胞24h后,用Western Blot和Real-Time PCR检测TGF-β1、CTGF、FN的表达,与对照组比较,TGF-β1、CTGF、FN的表达量均明显增加,并呈剂量依赖性。加入PGE1后,与单纯尿酸组比较,TGF-β1、CTGF、FN表达量下降明显。
结论：①.尿酸抑制HK-2细胞增殖和促进凋亡,增加NAG酶的释放；②.尿酸促HK-2细胞纤维化相关因子的表达；③.PGE1能降低尿酸所致HK-2细胞凋亡,降低尿酸诱导纤维化相关因子在HK-2细胞中的表达。
第二部分尿素对人肾小管上皮细胞的凋亡、纤维化相关因子表达的影响及PGE1干预的体外研究
目的：研究尿素对人肾小管上皮细胞的凋亡、纤维化相关因子表达的影响以及PGE1的干预作用。
方法：①.培养HK-2细胞；②.不同浓度的尿素刺激HK-2细胞24h、48h、72h,用MTT比色法检测细胞活性抑制率；用不同浓度尿素培养HK-2细胞24h,检测NAG酶的释放量及Annexin V.FITC/PI流式细胞检测凋亡来观察尿素对HK-2细胞的毒性作用；通过Western Blot和Real-Time PCR检测’TGF-β1、CTGF、FN的表达,观察尿素对HK-2细胞的促纤维化作用；并加入PGE1了解其对HK-2细胞的保护作用。
结果：①.HK-2细胞加入不同浓度的尿素分别作用24h、48h、72h后,与对照组比较,细胞活性抑制率明显升高,呈剂量依赖性。尿素作用HK-2细胞24h后,与对照组比较,NAG酶释放量呈剂量依赖性增加至14.23±0.750,细胞凋亡率也呈剂量依赖性增加至42.97%±3.012%; 160mmol/L尿素组加入PGE1后,与160mmol/L尿素组比较,细胞活性抑制率下降明显,NAG酶释放量明显减少至10.603±0.698,细胞凋亡率也明显降低至3531%±2.308%,均有显著性差异；②.用不同浓度的尿素作用HK-2细胞24h后,用Western Blot和Real-Time PCR检测TGF-β1、CTGF、FN的表达,与对照组比较,TGF-β1、CTGF、FN的表达量均明显增加,并呈剂量依赖性。加入PGE1后,与单纯尿素组比较,TGF-β1、CTGF、FN表达量下降明显。
结论：①.尿素抑制HK-2细胞增殖,促进凋亡,增加NAG酶的释放；②.尿素促纤维化相关因子TGF-β1、CTGF、FN在HK-2细胞中的表达；⑧.PGE1能降低尿素所致HK-2细胞凋亡,降低尿素诱导纤维化相关因子在HK-2细胞中的表达。
Part 1 Effect of uric acid on HK-2 cells apoptosis and expression of fibrotic correlation factors and intervention of PGE1
Objective:The study was designed to investigate the effect of UA on renal proximal tubular cells apoptosis and expression of fibrotic correlation factors as well as the protective effect of PGE1 against Urea-induced apoptosis and expression of fibrotic correlation factors in renal proximal tubular cells。
Methods:1. HK-2 cells were used as the subject;2.The toxic effect of UA on HK-2 cells was determined by MTT assay、NAG activity and Annexin V.FITC/PI fluorescence.3.Western Blot and Real-Time PCR were employed to investigate the expression of TGF-β1, CTGF and FN. In the blocking study, PGE1 was used as antagonists respectively.
Results:HK-2 Cells were exposed to UA (100,200,400, 800umol/L) respectively for 24h,48h,72h. UA could induce a significant dose and time-dependent loss of cell viability. Have been treated with different concentrations UA for 24h, NAG concentration and apoptosis ratio were increased dose dependently (P<0.01). The presence of PGE1 significantly lowered the inhibition rate of cell activity, NAG concentration and apoptotic ratio compared with the cells which were treated with Urea only.The protein and mRNA of TGF-β1,CTGF,FN in HK-2 cells increased significantly when HK-2 cells were cultivated with Urea, when PGE1 was added in UA fibrotic correlation factors decrease significantly.
Conclusion:1.UA could inhibition of renal proximal tubular cells proliferation and promote apoptosis in vitro; 2.UA could promote the expression of fibrosis relation factor(TGF-(31,CTGF,FN)in renal tubular epithelial cells; 3. PGE1 can interrupt apoptosis and the expression of fibrosis correlation factor caused by UA to HK-2 cells.
Part II Effect of urea on HK-2 cells apoptosis and expression of fibrotic correlation factors and intervention of PGE1
Objective:The study was designed to investigate the effect of urea on renal proximal tubular cells apoptosis and expression of fibrotic correlation factors as well as the protective effect of PGE1 against Urea-induced apoptosis and expression of fibrotic correlation factors in renal proximal tubular cells
Methods:1. HK-2 cells were used as the subject;2.The toxic effect of urea on HK-2 cells was determined by MTT assay、NAG activity and Annexin V-FTC/PI fluorescence; 3.Western Blot and Real-Time PCR were employed to investigate the expression of TGF-β1, CTGF and FN. In the blocking study, PGE1 was used as antagonists respectively.
Results:HK-2 Cells were exposed to urea (100,200,400 or 800umol/L) respectively for 24h,48h,72h. Urea could induce a significant dose and time-dependent loss of cell viability. Have been treated with different concentrations Urea for 24h, NAG concentration and apoptosis ratio were increased dose dependently (P<0.01). The presence of PGE1 significantly lowered the inhibition rate of cell activity, NAG concentration and apoptotic ratio compared with the cells which were treated with Urea only.The protein and mRNA of TGF-β1, CTGF, FN in HK-2 cells increased significantly when HK-2 cells were cultivated with Urea, when PGE1 wad added in Urea fibrotic correlation factors decrease significantly.
Conclusion:1.Urea could inhibition of renal proximal tubular cells proliferation and promote apoptosis in vitro; 2.Urea could promote the expression of fibrosis relation factor (TGF-β1,CTGF,FN)in renal tubular epithelial cells; 3. PGE1 can interrupt apoptosis and the expression of fibrosis correlation factor caused by Urea to HK-2 cells.
目的：研究尿酸对人肾小管上皮细胞凋亡、纤维化相关因子表达的影响以及PGE1的干预作用。
方法：①.培养HK-2细胞；②.不同浓度的尿酸刺激HK-2细胞24h、48h、72h,用MTT比色法检测细胞活性抑制率；不同浓度尿酸培养HK-2细胞24h,用Annexin V. FITC/PI流式细胞检测凋亡及检测NAG酶的释放量,来观察尿酸对HK-2细胞的毒性作用；通过Western Blot和Real-Time PCR检测TGF-β1、CTGF、FN的表达,观察尿酸对HK-2细胞的促纤维化作用；并加入PGE1了解其对HK-2细胞的保护作用。
结果：①.HK-2细胞加入不同浓度的尿酸分别作用24h、48h、72h后,与对照组比较,细胞活性抑制率明显升高,并呈时间剂量依赖性。尿酸作用HK-2细胞24h后,与对照组比较,细胞凋亡率呈剂量依赖性增加至56.06%±4.937%,NAG酶释放量也呈剂量依赖性增加至15.23±0.617。800umol/L尿酸组加入PGE1后,与800umol/L尿酸组比较,细胞活性抑制率下降明显,细胞凋亡率也明显降低至41.61%±4.149%,NAG酶释放量明显减少至11.24±0.468。②.用不同浓度的尿酸作用HK-2细胞24h后,用Western Blot和Real-Time PCR检测TGF-β1、CTGF、FN的表达,与对照组比较,TGF-β1、CTGF、FN的表达量均明显增加,并呈剂量依赖性。加入PGE1后,与单纯尿酸组比较,TGF-β1、CTGF、FN表达量下降明显。
结论：①.尿酸抑制HK-2细胞增殖和促进凋亡,增加NAG酶的释放；②.尿酸促HK-2细胞纤维化相关因子的表达；③.PGE1能降低尿酸所致HK-2细胞凋亡,降低尿酸诱导纤维化相关因子在HK-2细胞中的表达。
第二部分尿素对人肾小管上皮细胞的凋亡、纤维化相关因子表达的影响及PGE1干预的体外研究
目的：研究尿素对人肾小管上皮细胞的凋亡、纤维化相关因子表达的影响以及PGE1的干预作用。
方法：①.培养HK-2细胞；②.不同浓度的尿素刺激HK-2细胞24h、48h、72h,用MTT比色法检测细胞活性抑制率；用不同浓度尿素培养HK-2细胞24h,检测NAG酶的释放量及Annexin V.FITC/PI流式细胞检测凋亡来观察尿素对HK-2细胞的毒性作用；通过Western Blot和Real-Time PCR检测’TGF-β1、CTGF、FN的表达,观察尿素对HK-2细胞的促纤维化作用；并加入PGE1了解其对HK-2细胞的保护作用。
结果：①.HK-2细胞加入不同浓度的尿素分别作用24h、48h、72h后,与对照组比较,细胞活性抑制率明显升高,呈剂量依赖性。尿素作用HK-2细胞24h后,与对照组比较,NAG酶释放量呈剂量依赖性增加至14.23±0.750,细胞凋亡率也呈剂量依赖性增加至42.97%±3.012%; 160mmol/L尿素组加入PGE1后,与160mmol/L尿素组比较,细胞活性抑制率下降明显,NAG酶释放量明显减少至10.603±0.698,细胞凋亡率也明显降低至3531%±2.308%,均有显著性差异；②.用不同浓度的尿素作用HK-2细胞24h后,用Western Blot和Real-Time PCR检测TGF-β1、CTGF、FN的表达,与对照组比较,TGF-β1、CTGF、FN的表达量均明显增加,并呈剂量依赖性。加入PGE1后,与单纯尿素组比较,TGF-β1、CTGF、FN表达量下降明显。
结论：①.尿素抑制HK-2细胞增殖,促进凋亡,增加NAG酶的释放；②.尿素促纤维化相关因子TGF-β1、CTGF、FN在HK-2细胞中的表达；⑧.PGE1能降低尿素所致HK-2细胞凋亡,降低尿素诱导纤维化相关因子在HK-2细胞中的表达。
Part 1 Effect of uric acid on HK-2 cells apoptosis and expression of fibrotic correlation factors and intervention of PGE1
Objective:The study was designed to investigate the effect of UA on renal proximal tubular cells apoptosis and expression of fibrotic correlation factors as well as the protective effect of PGE1 against Urea-induced apoptosis and expression of fibrotic correlation factors in renal proximal tubular cells。
Methods:1. HK-2 cells were used as the subject;2.The toxic effect of UA on HK-2 cells was determined by MTT assay、NAG activity and Annexin V.FITC/PI fluorescence.3.Western Blot and Real-Time PCR were employed to investigate the expression of TGF-β1, CTGF and FN. In the blocking study, PGE1 was used as antagonists respectively.
Results:HK-2 Cells were exposed to UA (100,200,400, 800umol/L) respectively for 24h,48h,72h. UA could induce a significant dose and time-dependent loss of cell viability. Have been treated with different concentrations UA for 24h, NAG concentration and apoptosis ratio were increased dose dependently (P<0.01). The presence of PGE1 significantly lowered the inhibition rate of cell activity, NAG concentration and apoptotic ratio compared with the cells which were treated with Urea only.The protein and mRNA of TGF-β1,CTGF,FN in HK-2 cells increased significantly when HK-2 cells were cultivated with Urea, when PGE1 was added in UA fibrotic correlation factors decrease significantly.
Conclusion:1.UA could inhibition of renal proximal tubular cells proliferation and promote apoptosis in vitro; 2.UA could promote the expression of fibrosis relation factor(TGF-(31,CTGF,FN)in renal tubular epithelial cells; 3. PGE1 can interrupt apoptosis and the expression of fibrosis correlation factor caused by UA to HK-2 cells.
Part II Effect of urea on HK-2 cells apoptosis and expression of fibrotic correlation factors and intervention of PGE1
Objective:The study was designed to investigate the effect of urea on renal proximal tubular cells apoptosis and expression of fibrotic correlation factors as well as the protective effect of PGE1 against Urea-induced apoptosis and expression of fibrotic correlation factors in renal proximal tubular cells
Methods:1. HK-2 cells were used as the subject;2.The toxic effect of urea on HK-2 cells was determined by MTT assay、NAG activity and Annexin V-FTC/PI fluorescence; 3.Western Blot and Real-Time PCR were employed to investigate the expression of TGF-β1, CTGF and FN. In the blocking study, PGE1 was used as antagonists respectively.
Results:HK-2 Cells were exposed to urea (100,200,400 or 800umol/L) respectively for 24h,48h,72h. Urea could induce a significant dose and time-dependent loss of cell viability. Have been treated with different concentrations Urea for 24h, NAG concentration and apoptosis ratio were increased dose dependently (P<0.01). The presence of PGE1 significantly lowered the inhibition rate of cell activity, NAG concentration and apoptotic ratio compared with the cells which were treated with Urea only.The protein and mRNA of TGF-β1, CTGF, FN in HK-2 cells increased significantly when HK-2 cells were cultivated with Urea, when PGE1 wad added in Urea fibrotic correlation factors decrease significantly.
Conclusion:1.Urea could inhibition of renal proximal tubular cells proliferation and promote apoptosis in vitro; 2.Urea could promote the expression of fibrosis relation factor (TGF-β1,CTGF,FN)in renal tubular epithelial cells; 3. PGE1 can interrupt apoptosis and the expression of fibrosis correlation factor caused by Urea to HK-2 cells.
引文
[1]Dell, K, et al, Impact of PGE1 on cyclosporine A induced up-regulation of TGF-betal, its receptors, and related matrix production in cultured mesangial cells. Cytokine,2003.22(6):p.189-93.
[2]Anzai, N, Y. Kanai, and H. Endou, New insights into renal transport of urate. Curr Opin Rheumatol,2007.19(2):p.151-7.
[3]胡大一、丁荣晶等,无症状高尿酸血症合并心血管疾病诊治建议中国专家共识.中国全科医学,2010.11.
[4]Jossa, F, et al, Serum uric acid and hypertension:the Olivetti heart study. J Hum Hypertens,1994.8(9):p.677-81.
[5]Feig, D.I., D.H. Kang, and R.J. Johnson, Uric acid and cardiovascular risk. N Engl J Med,2008.359(17):p.1811-21.
[6]Bickel, C, et al, Serum uric acid as an independent predictor of mortality in patients with angiographically proven coronary artery disease. Am J Cardiol, 2002.89(1):p.12-7.
[7]Beevers, D.G. and GY. Lip, Is uric acid really an independent cardiovascular risk factor? Lancet,1998.352(9139):p.1556.
[8]DH, K. N. T, and F. L, A role for uric acid in the progression of renal disease. J Am Soc Nephro,2002.13:p.2888-2897.
[9]Kanellis, J, et al, Uric acid stimulates monocyte chemoattractant protein-1 production in vascular smooth muscle cells via mitogen-activated protein kinase and cyclooxygenase-2. Hypertension,2003.41(6):p.1287-93.
[10]Mazzali, M., et al, Elevated uric acid increases blood pressure in the rat by a novel crystal-independent mechanism. Hypertension,2001.38(5):p.1101-6.
[11]Gersch, C, et al, Inactivation of nitric oxide by uric acid. Nucleosides Nucleotides Nucleic Acids,2008.27(8):p.967-78.
[12]Khosla, U.M, et al, Hyperuricemia induces endothelial dysfunction. Kidney Int, 2005.67(5):p.1739-42.
[13]Madero, M, et al, Uric acid and long-term outcomes in CKD. Am J Kidney Dis, 2009.53(5):p.796-803.
[14]Turgut, F, B. Kasapoglu, and M. Kanbay, Uric acid, cardiovascular mortality, and long-term outcomes in CKD. Am J Kidney Dis,2009.54(3):p.582; author reply 582-3.
[15]Marangella, M, Uric acid elimination in the urine. Pathophysiological implications. Contrib Nephrol,2005.147:p.132-48.
[16]Weiner, D.E, et al, Uric acid and incident kidney disease in the community. J Am Soc Nephrol,2008.19(6):p.1204-11.
[17]Iseki, K, et al, Significance of hyperuricemia on the early detection of renal failure in a cohort of screened subjects. Hypertens Res,2001.24(6):p.691-7.
[18]Iseki, K, et al, Significance of hyperuricemia as a risk factor for developing ESRD in a screened cohort. Am J Kidney Dis,2004.44(4):p.642-50.
[19]Chonchol, M, et al, Relationship of uric acid with progression of kidney disease. Am J Kidney Dis,2007.50(2):p.239-47.
[20]Bellomo, G, et al, Association of uric acid with change in kidney function in healthy normotensive individuals. Am J Kidney Dis,2010.56(2):p.264-72.
[21]Obermayr, R.P, et al, Elevated uric acid increases the risk for kidney disease. J Am Soc Nephrol,2008.19(12):p.2407-13.
[22]邱强、陈香美等,影响IgA肾病高尿酸血症的因素.中国中西医结合肾脏病杂志,2005.6(6)：p.6.
[23]Cameron, J.S. and H.A. Simmonds, Uric acid, gout and the kidney. J Clin Pathol,1981.34(11):p.1245-54.
[24]Terkeltaub, R.A, Gout and mechanisms of crystal-induced inflammation. Curr Opin Rheumatol,1993.5(4):p.510-6.
[25]Iaina, A. and D. Schwartz, Renal tubular cellular and molecular events in acute renal failure. Nephron,1994.68(4):p.413-8.
[26]Wang, Y, et al, TGF-betal/Smad7 signaling stimulates renal tubulointerstitial fibrosis induced by AAI. J Recept Signal Transduct Res,2008.28(4):p. 413-28.
[27]Cheng, O, et al, Connective tissue growth factor is a biomarker and mediator of kidney allograft fibrosis. Am J Transplant,2006.6(10):p.2292-306.
[28]Burns, W.C, et al, Connective tissue growth factor plays an important role in advanced glycation end product-induced tubular epithelial-to-mesenchymal transition:implications for diabetic renal disease. J Am Soc Nephrol,2006. 17(9):p.2484-94.
[29]Serini, G, et al, The fibronectin domain ED-A is crucial for myofibroblastic phenotype induction by transforming growth factor-betal. J Cell Biol,1998. 142(3):p.873-81.
[30]Gharaee-Kermani. M, et al, Fibronectin is the major fibroblast chemoattractant in rabbit anti-glomerular basement membrane disease. Am J Pathol,1996. 148(3):p.961-7.
[31]Ulmasov, B, et al, Protective role of angiotensin Ⅱ type 2 receptor signaling in a mouse model of pancreatic fibrosis. Am J Physiol Gastrointest Liver Physiol, 2009.296(2):p. G284-94.
[32]刘超等,洛伐他汀对人肾小球系膜细胞转化生长因子-β1和细胞外基质的影响.实用临床医药杂志,2004.8：p.322-326.
[33]Panichi.V,et al, Effects of 1,25-(OH)2D3 in experimental mesangial proliferative nephritis in rats. Kidney Int,2001.60(1):p.87-95.
[34]Hsu,Y.H,et al,Prostacyclin protects renal tubular cells from gentamicin-induced apoptosis via a PPARalpha-dependent pathway. Kidney Int,2008.73(5):p. 578-87.
[35]Tamura, D.Y, et al, Prostaglandin E1 attenuates cytotoxic mechanisms of primed neutrophils. Shock,1998.9(3):p.171-6.
[36]Matlhagela, K. and M. Taub, Regulation of the Na-K-ATPase beta(1)-subunit promoter by multiple prostaglandin-responsive elements. Am J Physiol Renal Physiol,2006.291(3):p. F635-46.
[37]Vanholder, R, et al, Review on uremic toxins:classification, concentration, and interindividual variability. Kidney Int,2003.63(5):p.1934-43.
[38]Vanholder, R, et al, Review on uraemic solutes Ⅱ--variability in reported concentrations:causes and consequences. Nephrol Dial Transplant,2007. 22(11):p.3115-21.
[39]Cohen, G, et al, Review on uraemic toxins Ⅲ:recommendations for handling uraemic retention solutes in vitro--towards a standardized approach for research on uraemia. Nephrol Dial Transplant,2007.22(12):p.3381-90.
[40]Jamison, R.L, et al, Effect of homocysteine lowering on mortality and vascular disease in advanced chronic kidney disease and end-stage renal disease:a randomized controlled trial. JAMA,2007.298(10):p.1163-70.
[41]Kumagai, H, et al, Renal tubulointerstitial injury in weanling rats with hyperhomocysteinemia. Kidney Int,2002.62(4):p.1219-28.
[42]Chow, F.Y, et al, Intercellular adhesion molecule-1 deficiency is protective against nephropathy in type 2 diabetic db/db mice. J Am Soc Nephrol,2005. 16(6):p.1711-22.
[43]Schmidt, A.M, et al, Activation of receptor for advanced glycation end products: a mechanism for chronic vascular dysfunction in diabetic vasculopathy and atherosclerosis. Circ Res,1999.84(5):p.489-97.
[44]Walser, M. and L.J. Bodenlos, Urea metabolism in man. J Clin Invest,1959.38: p.1617-26.
[45]Walser, M., Urea metabolism in chronic renal failure. J Clin Invest,1974.53(5): p.1385-92.
[46]Long, C.L, M. Jeevanandam, and J.M. Kinney, Metabolism and recycling of urea in man. Am J Clin Nutr,1978.31(8):p.1367-82.
[47]Wang, Y, et al, Epac regulates UT-A1 to increase urea transport in inner medullary collecting ducts. J Am Soc Nephrol,2009.20(9):p.2018-24.
[48]Himmelfarb, J, Urea:surrogate or toxin? Kidney Int,1999.56(2):p.754-5.
[49]Johnson, W.J. et al., Effects of urea loading in patients with far-advanced renal failure. Mayo Clin Proc,1972.47(1):p.21-9.
[50]Lee, J.A. H.A. Lee, and P.J. Sadler, Uraemia:is urea more important than we think? Lancet,1991.338(8780):p.1438-40.
[51]Grollman, E.F. and A. Grollman, Toxicity of urea and its role in the pathogenesis of uremia. J Clin Invest,1959.38(5):p.749-54.
[52]HW, S, The physiology of the kidney.1937:p.1-310.
[53]Mun, K.C. and T.A. Golper, Impaired biological activity of erythropoietin by cyanate carbamylation. Blood Purif,2000.18(1):p.13-7.
[54]Kraus, L.M. and A.P. Kraus, Jr. Carbamoylation of amino acids and proteins in uremia. Kidney Int Suppl,2001.78:p. S102-7.
[55]Kjellstrand, C.M, et al, Short daily haemodialysis:survival in 415 patients treated for 1006 patient-years. Nephrol Dial Transplant,2008.23(10):p. 3283-9.
[56]Michea, L, et al, Cell cycle delay and apoptosis are induced by high salt and urea in renal medullary cells. Am J Physiol Renal Physiol,2000.278(2):p. F209-18.
[57]Zhang, Z, et al, High urea and NaC1 carbonylate proteins in renal cells in culture and in vivo, and high urea causes 8-oxoguanine lesions in their DNA. Proc Natl Acad Sci U S A,2004.101(25):p.9491-6.
[58]Moeslinger, T, et al, Urea induces macrophage proliferation by inhibition of inducible nitric oxide synthesis. Kidney Int,1999.56(2):p.581-8.
[59]Prabhakar, S.S, et al,Urea inhibits inducible nitric oxide synthase in macrophage cell line. Am J Physiol,1997.273(6 Pt 1):p. C1882-8.
[60]D'Apolito, M, et al. Urea-induced ROS generation causes insulin resistance in mice with chronic renal failure. J Clin Invest,2010.120(1):p.203-13.
[61]Eddy, A.A. et al. Molecular basis of renal fibrosis. Pediatr Nephrol,2000. 15(3-4):p.290-301.
[62]Sanchez-Lopez, E, et al, CTGF promotes inflammatory cell infiltration of the renal interstitium by activating NF-kappaB. J Am Soc Nephrol,2009.20(7):p. 1513-26.
[63]Yokoi, H, et al, Role of connective tissue growth factor in fibronectin expression and tubulointerstitial fibrosis. Am J Physiol Renal Physiol,2002. 282(5):p. F933-42.
[64]Chen, L, et al, Influence of connective tissue growth factor antisense oligonucleotide on angiotensin Ⅱ-induced epithelial mesenchymal transition in HK2 cells. Acta Pharmacol Sin,2006.27(8):p.1029-36.
[1]Richet, G, Early history of uremia. Kidney Int,1988.33(5):p.1013-5.
[2]Vanholder, R., et al, Review on uraemic solutes Ⅱ--variability in reported concentrations:causes and consequences. Nephrol Dial Transplant,2007. 22(11):p.3115-21.
[3]Cohen, G., et al, Review on uraemic toxins III:recommendations for handling uraemic retention solutes in vitro--towards a standardized approach for research on uraemia. Nephrol Dial Transplant,2007.22(12):p.3381-90.
[4]Wiesholzer, M., et al, Inappropriately high plasma leptin levels in obese haemodialysis patients can be reduced by high flux haemodialysis and haemodiafiltration. Clin Sci (Lond),1998.94(4):p.431-5.
[5]McCully, K.S. and R.B. Wilson, Homocysteine theory of arteriosclerosis. Atherosclerosis,1975.22(2):p.215-27.
[6]Kumagai, H., et al, Renal tubulointerstitial injury in weanling rats with hyperhomocysteinemia. Kidney Int,2002.62(4):p.1219-28.
[7]Li, N., Y.F. Chen, and A.P. Zou, Implications of hyperhomocysteinemia in glomerular sclerosis in hypertension. Hypertension,2002.39(2 Pt 2):p.443-8.
[8]Schmidt, A.M., et al, Activation of receptor for advanced glycation end products:a mechanism for chronic vascular dysfunction in diabetic vasculopathy and atherosclerosis. Circ Res,1999.84(5):p.489-97.
[9]Chow, F.Y, et al, Intercellular adhesion molecule-1 deficiency is protective against nephropathy in type 2 diabetic db/db mice. J Am Soc Nephrol,2005. 16(6):p.1711-22.
[10]Guh, J.Y, et al, Advanced glycation end product-induced proliferation in NRK-49F cells is dependent on the JAK2/STAT5 pathway and cyclin D1. Am J Kidney Dis,2001.38(5):p.1096-104.
[11]Han, D.C, et al, Leptin stimulates type I collagen production in db/db mesangial cells:glucose uptake and TGF-beta type II receptor expression. Kidney Int,2001.59(4):p.1315-23.
[12]Ballermann, B.J, A role for leptin in glomerulosclerosis? Kidney Int,1999. 56(3):p.1154-5.
[13]Wolf, G, et al, Leptin and renal disease. Am J Kidney Dis,2002.39(1):p.1-11.
[14]Lee, C.I, et al, Leptin and connective tissue growth factor in advanced glycation end-product-induced effects in NRK-49F cells. J Cell Biochem,2004. 93(5):p.940-50.
[15]Miyazaki, T, et al, Indoxyl sulfate stimulates renal synthesis of transforming growth factor-beta 1 and progression of renal failure. Kidney Int Suppl,1997. 63:p. S211-4.
[16]Miyazaki, T, et al, Indoxyl sulfate increases the gene expressions of TGF-beta 1, TIMP-1 and pro-alpha 1(I) collagen in uremic rat kidneys. Kidney Int Suppl, 1997.62:p. S15-22.
[17]Motojima, M, et al, Uraemic toxins induce proximal tubular injury via organic anion transporter 1-mediated uptake. Br J Pharmacol,2002.135(2):p.555-63.
[18]Motojima, M, et al, Uremic toxins of organic anions up-regulate PAI-1 expression by induction of NF-kappaB and free radical in proximal tubular cells. Kidney Int,2003.63(5):p.1671-80.
[19]Okada, K, et al, Combination therapy with angiotensin-converting enzyme inhibitor and oral adsorbent of uremic toxins can delay the appearance of glomerular sclerosis and interstitial fibrosis in established renal failure. Kidney Blood Press Res,2004.27(4):p.218-25.
[20]D'Hooge, R, et al, Involvement of voltage-and ligand-gated Ca2+ channels in the neuroexcitatory and synergistic effects of putative uremic neurotoxins. Kidney Int,2003.63(5):p.1764-75.
[21]Wang, F, et al, Methylguanidine cytotoxicity on HK-2 cells and protective effect of antioxidants against MG-induced apoptosis in renal proximal tubular cells in vitro. Ren Fail.32(8):p.978-85.
[22]Winchester, J.F. and P.F. Audia, Extracorporeal strategies for the removal of middle molecules. Semin Dial,2006.19(2):p.110-4.
[2]Anzai, N, Y. Kanai, and H. Endou, New insights into renal transport of urate. Curr Opin Rheumatol,2007.19(2):p.151-7.
[3]胡大一、丁荣晶等,无症状高尿酸血症合并心血管疾病诊治建议中国专家共识.中国全科医学,2010.11.
[4]Jossa, F, et al, Serum uric acid and hypertension:the Olivetti heart study. J Hum Hypertens,1994.8(9):p.677-81.
[5]Feig, D.I., D.H. Kang, and R.J. Johnson, Uric acid and cardiovascular risk. N Engl J Med,2008.359(17):p.1811-21.
[6]Bickel, C, et al, Serum uric acid as an independent predictor of mortality in patients with angiographically proven coronary artery disease. Am J Cardiol, 2002.89(1):p.12-7.
[7]Beevers, D.G. and GY. Lip, Is uric acid really an independent cardiovascular risk factor? Lancet,1998.352(9139):p.1556.
[8]DH, K. N. T, and F. L, A role for uric acid in the progression of renal disease. J Am Soc Nephro,2002.13:p.2888-2897.
[9]Kanellis, J, et al, Uric acid stimulates monocyte chemoattractant protein-1 production in vascular smooth muscle cells via mitogen-activated protein kinase and cyclooxygenase-2. Hypertension,2003.41(6):p.1287-93.
[10]Mazzali, M., et al, Elevated uric acid increases blood pressure in the rat by a novel crystal-independent mechanism. Hypertension,2001.38(5):p.1101-6.
[11]Gersch, C, et al, Inactivation of nitric oxide by uric acid. Nucleosides Nucleotides Nucleic Acids,2008.27(8):p.967-78.
[12]Khosla, U.M, et al, Hyperuricemia induces endothelial dysfunction. Kidney Int, 2005.67(5):p.1739-42.
[13]Madero, M, et al, Uric acid and long-term outcomes in CKD. Am J Kidney Dis, 2009.53(5):p.796-803.
[14]Turgut, F, B. Kasapoglu, and M. Kanbay, Uric acid, cardiovascular mortality, and long-term outcomes in CKD. Am J Kidney Dis,2009.54(3):p.582; author reply 582-3.
[15]Marangella, M, Uric acid elimination in the urine. Pathophysiological implications. Contrib Nephrol,2005.147:p.132-48.
[16]Weiner, D.E, et al, Uric acid and incident kidney disease in the community. J Am Soc Nephrol,2008.19(6):p.1204-11.
[17]Iseki, K, et al, Significance of hyperuricemia on the early detection of renal failure in a cohort of screened subjects. Hypertens Res,2001.24(6):p.691-7.
[18]Iseki, K, et al, Significance of hyperuricemia as a risk factor for developing ESRD in a screened cohort. Am J Kidney Dis,2004.44(4):p.642-50.
[19]Chonchol, M, et al, Relationship of uric acid with progression of kidney disease. Am J Kidney Dis,2007.50(2):p.239-47.
[20]Bellomo, G, et al, Association of uric acid with change in kidney function in healthy normotensive individuals. Am J Kidney Dis,2010.56(2):p.264-72.
[21]Obermayr, R.P, et al, Elevated uric acid increases the risk for kidney disease. J Am Soc Nephrol,2008.19(12):p.2407-13.
[22]邱强、陈香美等,影响IgA肾病高尿酸血症的因素.中国中西医结合肾脏病杂志,2005.6(6)：p.6.
[23]Cameron, J.S. and H.A. Simmonds, Uric acid, gout and the kidney. J Clin Pathol,1981.34(11):p.1245-54.
[24]Terkeltaub, R.A, Gout and mechanisms of crystal-induced inflammation. Curr Opin Rheumatol,1993.5(4):p.510-6.
[25]Iaina, A. and D. Schwartz, Renal tubular cellular and molecular events in acute renal failure. Nephron,1994.68(4):p.413-8.
[26]Wang, Y, et al, TGF-betal/Smad7 signaling stimulates renal tubulointerstitial fibrosis induced by AAI. J Recept Signal Transduct Res,2008.28(4):p. 413-28.
[27]Cheng, O, et al, Connective tissue growth factor is a biomarker and mediator of kidney allograft fibrosis. Am J Transplant,2006.6(10):p.2292-306.
[28]Burns, W.C, et al, Connective tissue growth factor plays an important role in advanced glycation end product-induced tubular epithelial-to-mesenchymal transition:implications for diabetic renal disease. J Am Soc Nephrol,2006. 17(9):p.2484-94.
[29]Serini, G, et al, The fibronectin domain ED-A is crucial for myofibroblastic phenotype induction by transforming growth factor-betal. J Cell Biol,1998. 142(3):p.873-81.
[30]Gharaee-Kermani. M, et al, Fibronectin is the major fibroblast chemoattractant in rabbit anti-glomerular basement membrane disease. Am J Pathol,1996. 148(3):p.961-7.
[31]Ulmasov, B, et al, Protective role of angiotensin Ⅱ type 2 receptor signaling in a mouse model of pancreatic fibrosis. Am J Physiol Gastrointest Liver Physiol, 2009.296(2):p. G284-94.
[32]刘超等,洛伐他汀对人肾小球系膜细胞转化生长因子-β1和细胞外基质的影响.实用临床医药杂志,2004.8：p.322-326.
[33]Panichi.V,et al, Effects of 1,25-(OH)2D3 in experimental mesangial proliferative nephritis in rats. Kidney Int,2001.60(1):p.87-95.
[34]Hsu,Y.H,et al,Prostacyclin protects renal tubular cells from gentamicin-induced apoptosis via a PPARalpha-dependent pathway. Kidney Int,2008.73(5):p. 578-87.
[35]Tamura, D.Y, et al, Prostaglandin E1 attenuates cytotoxic mechanisms of primed neutrophils. Shock,1998.9(3):p.171-6.
[36]Matlhagela, K. and M. Taub, Regulation of the Na-K-ATPase beta(1)-subunit promoter by multiple prostaglandin-responsive elements. Am J Physiol Renal Physiol,2006.291(3):p. F635-46.
[37]Vanholder, R, et al, Review on uremic toxins:classification, concentration, and interindividual variability. Kidney Int,2003.63(5):p.1934-43.
[38]Vanholder, R, et al, Review on uraemic solutes Ⅱ--variability in reported concentrations:causes and consequences. Nephrol Dial Transplant,2007. 22(11):p.3115-21.
[39]Cohen, G, et al, Review on uraemic toxins Ⅲ:recommendations for handling uraemic retention solutes in vitro--towards a standardized approach for research on uraemia. Nephrol Dial Transplant,2007.22(12):p.3381-90.
[40]Jamison, R.L, et al, Effect of homocysteine lowering on mortality and vascular disease in advanced chronic kidney disease and end-stage renal disease:a randomized controlled trial. JAMA,2007.298(10):p.1163-70.
[41]Kumagai, H, et al, Renal tubulointerstitial injury in weanling rats with hyperhomocysteinemia. Kidney Int,2002.62(4):p.1219-28.
[42]Chow, F.Y, et al, Intercellular adhesion molecule-1 deficiency is protective against nephropathy in type 2 diabetic db/db mice. J Am Soc Nephrol,2005. 16(6):p.1711-22.
[43]Schmidt, A.M, et al, Activation of receptor for advanced glycation end products: a mechanism for chronic vascular dysfunction in diabetic vasculopathy and atherosclerosis. Circ Res,1999.84(5):p.489-97.
[44]Walser, M. and L.J. Bodenlos, Urea metabolism in man. J Clin Invest,1959.38: p.1617-26.
[45]Walser, M., Urea metabolism in chronic renal failure. J Clin Invest,1974.53(5): p.1385-92.
[46]Long, C.L, M. Jeevanandam, and J.M. Kinney, Metabolism and recycling of urea in man. Am J Clin Nutr,1978.31(8):p.1367-82.
[47]Wang, Y, et al, Epac regulates UT-A1 to increase urea transport in inner medullary collecting ducts. J Am Soc Nephrol,2009.20(9):p.2018-24.
[48]Himmelfarb, J, Urea:surrogate or toxin? Kidney Int,1999.56(2):p.754-5.
[49]Johnson, W.J. et al., Effects of urea loading in patients with far-advanced renal failure. Mayo Clin Proc,1972.47(1):p.21-9.
[50]Lee, J.A. H.A. Lee, and P.J. Sadler, Uraemia:is urea more important than we think? Lancet,1991.338(8780):p.1438-40.
[51]Grollman, E.F. and A. Grollman, Toxicity of urea and its role in the pathogenesis of uremia. J Clin Invest,1959.38(5):p.749-54.
[52]HW, S, The physiology of the kidney.1937:p.1-310.
[53]Mun, K.C. and T.A. Golper, Impaired biological activity of erythropoietin by cyanate carbamylation. Blood Purif,2000.18(1):p.13-7.
[54]Kraus, L.M. and A.P. Kraus, Jr. Carbamoylation of amino acids and proteins in uremia. Kidney Int Suppl,2001.78:p. S102-7.
[55]Kjellstrand, C.M, et al, Short daily haemodialysis:survival in 415 patients treated for 1006 patient-years. Nephrol Dial Transplant,2008.23(10):p. 3283-9.
[56]Michea, L, et al, Cell cycle delay and apoptosis are induced by high salt and urea in renal medullary cells. Am J Physiol Renal Physiol,2000.278(2):p. F209-18.
[57]Zhang, Z, et al, High urea and NaC1 carbonylate proteins in renal cells in culture and in vivo, and high urea causes 8-oxoguanine lesions in their DNA. Proc Natl Acad Sci U S A,2004.101(25):p.9491-6.
[58]Moeslinger, T, et al, Urea induces macrophage proliferation by inhibition of inducible nitric oxide synthesis. Kidney Int,1999.56(2):p.581-8.
[59]Prabhakar, S.S, et al,Urea inhibits inducible nitric oxide synthase in macrophage cell line. Am J Physiol,1997.273(6 Pt 1):p. C1882-8.
[60]D'Apolito, M, et al. Urea-induced ROS generation causes insulin resistance in mice with chronic renal failure. J Clin Invest,2010.120(1):p.203-13.
[61]Eddy, A.A. et al. Molecular basis of renal fibrosis. Pediatr Nephrol,2000. 15(3-4):p.290-301.
[62]Sanchez-Lopez, E, et al, CTGF promotes inflammatory cell infiltration of the renal interstitium by activating NF-kappaB. J Am Soc Nephrol,2009.20(7):p. 1513-26.
[63]Yokoi, H, et al, Role of connective tissue growth factor in fibronectin expression and tubulointerstitial fibrosis. Am J Physiol Renal Physiol,2002. 282(5):p. F933-42.
[64]Chen, L, et al, Influence of connective tissue growth factor antisense oligonucleotide on angiotensin Ⅱ-induced epithelial mesenchymal transition in HK2 cells. Acta Pharmacol Sin,2006.27(8):p.1029-36.
[1]Richet, G, Early history of uremia. Kidney Int,1988.33(5):p.1013-5.
[2]Vanholder, R., et al, Review on uraemic solutes Ⅱ--variability in reported concentrations:causes and consequences. Nephrol Dial Transplant,2007. 22(11):p.3115-21.
[3]Cohen, G., et al, Review on uraemic toxins III:recommendations for handling uraemic retention solutes in vitro--towards a standardized approach for research on uraemia. Nephrol Dial Transplant,2007.22(12):p.3381-90.
[4]Wiesholzer, M., et al, Inappropriately high plasma leptin levels in obese haemodialysis patients can be reduced by high flux haemodialysis and haemodiafiltration. Clin Sci (Lond),1998.94(4):p.431-5.
[5]McCully, K.S. and R.B. Wilson, Homocysteine theory of arteriosclerosis. Atherosclerosis,1975.22(2):p.215-27.
[6]Kumagai, H., et al, Renal tubulointerstitial injury in weanling rats with hyperhomocysteinemia. Kidney Int,2002.62(4):p.1219-28.
[7]Li, N., Y.F. Chen, and A.P. Zou, Implications of hyperhomocysteinemia in glomerular sclerosis in hypertension. Hypertension,2002.39(2 Pt 2):p.443-8.
[8]Schmidt, A.M., et al, Activation of receptor for advanced glycation end products:a mechanism for chronic vascular dysfunction in diabetic vasculopathy and atherosclerosis. Circ Res,1999.84(5):p.489-97.
[9]Chow, F.Y, et al, Intercellular adhesion molecule-1 deficiency is protective against nephropathy in type 2 diabetic db/db mice. J Am Soc Nephrol,2005. 16(6):p.1711-22.
[10]Guh, J.Y, et al, Advanced glycation end product-induced proliferation in NRK-49F cells is dependent on the JAK2/STAT5 pathway and cyclin D1. Am J Kidney Dis,2001.38(5):p.1096-104.
[11]Han, D.C, et al, Leptin stimulates type I collagen production in db/db mesangial cells:glucose uptake and TGF-beta type II receptor expression. Kidney Int,2001.59(4):p.1315-23.
[12]Ballermann, B.J, A role for leptin in glomerulosclerosis? Kidney Int,1999. 56(3):p.1154-5.
[13]Wolf, G, et al, Leptin and renal disease. Am J Kidney Dis,2002.39(1):p.1-11.
[14]Lee, C.I, et al, Leptin and connective tissue growth factor in advanced glycation end-product-induced effects in NRK-49F cells. J Cell Biochem,2004. 93(5):p.940-50.
[15]Miyazaki, T, et al, Indoxyl sulfate stimulates renal synthesis of transforming growth factor-beta 1 and progression of renal failure. Kidney Int Suppl,1997. 63:p. S211-4.
[16]Miyazaki, T, et al, Indoxyl sulfate increases the gene expressions of TGF-beta 1, TIMP-1 and pro-alpha 1(I) collagen in uremic rat kidneys. Kidney Int Suppl, 1997.62:p. S15-22.
[17]Motojima, M, et al, Uraemic toxins induce proximal tubular injury via organic anion transporter 1-mediated uptake. Br J Pharmacol,2002.135(2):p.555-63.
[18]Motojima, M, et al, Uremic toxins of organic anions up-regulate PAI-1 expression by induction of NF-kappaB and free radical in proximal tubular cells. Kidney Int,2003.63(5):p.1671-80.
[19]Okada, K, et al, Combination therapy with angiotensin-converting enzyme inhibitor and oral adsorbent of uremic toxins can delay the appearance of glomerular sclerosis and interstitial fibrosis in established renal failure. Kidney Blood Press Res,2004.27(4):p.218-25.
[20]D'Hooge, R, et al, Involvement of voltage-and ligand-gated Ca2+ channels in the neuroexcitatory and synergistic effects of putative uremic neurotoxins. Kidney Int,2003.63(5):p.1764-75.
[21]Wang, F, et al, Methylguanidine cytotoxicity on HK-2 cells and protective effect of antioxidants against MG-induced apoptosis in renal proximal tubular cells in vitro. Ren Fail.32(8):p.978-85.
[22]Winchester, J.F. and P.F. Audia, Extracorporeal strategies for the removal of middle molecules. Semin Dial,2006.19(2):p.110-4.