三叶因子1在大鼠草酸钙结石模型肾组织中的表达及其意义
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摘要
目的
探讨三叶因子1(TFF1)在大鼠草酸钙结石模型肾组织中的表达与肾脏草酸钙结石形成机制的关系。
方法
SD大鼠分别给予1%的乙二醇溶液自由饮用和2%的NH4CI溶液2ml/d灌胃2周和4周；用普通光显微镜和相差显微镜观察结石结晶形成情况；采用免疫组织化学染色法分别检测成石2周组(n=20)、成石4周组n=20)和正常组(n=20)大鼠肾组织TFF1蛋白的表达。
结果
TFF1免疫反应阳性物质主要位于肾小球、肾小管。与正常组比较,成石2周组肾组织TFF1平均灰度值略下降(P>0.05),而成石4周组平均灰度值明显降低(P<0.05)。
结论
乙二醇诱导的大鼠草酸钙结石模型肾组织中TFF1低表达,伴随结石结晶数量和密度的增加,TFF1表达降低。TFF1在肾组织中的表达与乙二醇和NH4CI干预的时限呈负相关；TFF1可能在肾草酸钙结石的形成中起抑制作用。
Objective
To Explore the expression of trefoil factor (TFF1) in kidney tissue of rat with calcium oxalate stone and to investigate the association with the mechanism of renal calculi formation.
Method
Rats were raised continually through drinking1%ethylene glycol solution and gavaging2%NH4CI solution for2weeks and4weeks; kidney stone formation was observed using Polarizing microscope;and expression of TFF1in renal tissue was detected by immunohistochemistry in two weeks group (n=20), four weeks group (n=20) and normal group (n=20) respectively.
Results
TFFlimmunoreactive substances mainly located in the glomerular and tubular compared with normal group, the TFF1average gray value decreased slightly (P>0.05) in two weeks group and significantly reduced in four weeks group(P<0.05).
Conclusion
Expression of TFF1was reduced in kidney tissue of rat with calcium oxalate stone. Accompanied by the increase in the number and density of stone crystalline,TFF1expression decrease shows that TFF1is a inhibitor of calcium oxalate stone formation. High level oxalic acid and COM crystals can cause TFF1expression profile changes in rat kidney.The changes of TFF1expression may protect epithelial cell from being injured, and may be related to kidney stone formation
探讨三叶因子1(TFF1)在大鼠草酸钙结石模型肾组织中的表达与肾脏草酸钙结石形成机制的关系。
方法
SD大鼠分别给予1%的乙二醇溶液自由饮用和2%的NH4CI溶液2ml/d灌胃2周和4周；用普通光显微镜和相差显微镜观察结石结晶形成情况；采用免疫组织化学染色法分别检测成石2周组(n=20)、成石4周组n=20)和正常组(n=20)大鼠肾组织TFF1蛋白的表达。
结果
TFF1免疫反应阳性物质主要位于肾小球、肾小管。与正常组比较,成石2周组肾组织TFF1平均灰度值略下降(P>0.05),而成石4周组平均灰度值明显降低(P<0.05)。
结论
乙二醇诱导的大鼠草酸钙结石模型肾组织中TFF1低表达,伴随结石结晶数量和密度的增加,TFF1表达降低。TFF1在肾组织中的表达与乙二醇和NH4CI干预的时限呈负相关；TFF1可能在肾草酸钙结石的形成中起抑制作用。
Objective
To Explore the expression of trefoil factor (TFF1) in kidney tissue of rat with calcium oxalate stone and to investigate the association with the mechanism of renal calculi formation.
Method
Rats were raised continually through drinking1%ethylene glycol solution and gavaging2%NH4CI solution for2weeks and4weeks; kidney stone formation was observed using Polarizing microscope;and expression of TFF1in renal tissue was detected by immunohistochemistry in two weeks group (n=20), four weeks group (n=20) and normal group (n=20) respectively.
Results
TFFlimmunoreactive substances mainly located in the glomerular and tubular compared with normal group, the TFF1average gray value decreased slightly (P>0.05) in two weeks group and significantly reduced in four weeks group(P<0.05).
Conclusion
Expression of TFF1was reduced in kidney tissue of rat with calcium oxalate stone. Accompanied by the increase in the number and density of stone crystalline,TFF1expression decrease shows that TFF1is a inhibitor of calcium oxalate stone formation. High level oxalic acid and COM crystals can cause TFF1expression profile changes in rat kidney.The changes of TFF1expression may protect epithelial cell from being injured, and may be related to kidney stone formation
引文
1. Luqmani, Y.A., et al., An immunohistochemical survey of pS2 expression in human epithelial cancers. Int J Cancer,1992.50(2):p.302-4.
2. Boyce, W.H. and N.M. Sulkin, Biocolloids of urine in health and in calculous disease. Ⅲ. The mucoprotein matrix of urinary calculi. J Clin Invest,1956.35(10):p.1067-79.
3. Canales, B.K., et al., Proteome of human calcium kidney stones. Urology,2010.76(4):p.1017 e13-20.
4. 杜廷义.张勇,and张云,三叶因子：从实验室研究到临床医学.动物学研究.31(1)：p.11.
5. Masiakowski, P., et al., Cloning of cDNA sequences of hormone-regulated genes from the MCF-7 human breast cancer cell line. Nucleic Acids Res,1982.10(24):p.7895-903.
6. Terada, T., et al., Characterization of the mouse TFF1 (pS2) gene promoter region. Biol Pharm Bull,2001.24(2):p.135-9.
7. Thongboonkerd, V., Proteomics and kidney stone disease. Contrib Nephrol,2008.160:p. 142-58.
8. Kraus, S., et al., MUC1 mucin and trefoil factor 1 protein expression in renal cell carcinoma:correlation with prognosis. Hum Pathol,2002.33(1):p.60-7.
9. 蔡华芳,草酸钙肾结石模型及其应用中国中西医结合肾病杂志.11(8).
10. 赵宗江,et al.,筛选乙二醇法复制大鼠肾结石模型最佳剂量的比较研究.中国中西医结合肾病杂志,2007.8(11)：p.3.
11.厉建.李文峰.and 张士青.尿草酸对肾草钙结石形成和肾组织自由基影响的研究.中国实验诊断学,2008.12(7)：p.3.
12.钟玲.et.al.乙二醇法复制肾结石模型给药途径和方法的比较.中国病理生理杂志,2000.16(4)：p.2.
13. Hamamoto, S., K. Taguchi, and Y. Fujii, [Molecular mechanism of renal stone formation]. Clin Calcium.21(10):p.1481-7.
14. Liang, L., et al., Expression profiling of crystal-induced injury in human kidney epithelial cells. Nephron Physiol,2006.103(1):p. p53-62.
15. Khan, S.R., Role of renal epithelial cells in the initiation of calcium oxalate stones. Nephron Exp Nephrol,2004.98(2):p. e55-60.
16. Khand, F.D., et al., Mitochondrial superoxide production during oxalate-mediated oxidative stress in renal epithelial cells. Free Radic Biol Med,2002.32(12):p.1339-50.
17. Gokhale, J.A., M.D. McKee, and S.R. Khan, Immunocytochemical localization of Tamm-Horsfall protein in the kidneys of normal and nephrolithic rats. Urol Res,1996. 24(4):p.201-9.
18. Sejdiu, I. and O. Torffvit, Decreased urinary concentration of Tamm-Horsfall protein is associated with development of renal failure and cardiovascular death within 20 years in type 1 but not in type 2 diabetic patients. Scand J Urol Nephrol,2008.42(2):p.168-74.
19. Chakraborty, J., A.A. Below, and D. Solaiman, Tamm-Horsfall protein in patients with kidney damage and diabetes. Urol Res,2004.32(2):p.79-83.
20. Hess, B., L. Zipperle, and P. Jaeger, Citrate and calcium effects on Tamm-Horsfall glycoprotein as a modifier of calcium oxalate crystal aggregation. Am J Physiol,1993. 265(6 Pt 2):p. F784-91.
21. Rodriguez Camargo, D.C., N.M. Link, and S.A. Dames, The FKBP-rapamycin binding domain of human TOR undergoes strong conformational changes in the presence of membrane mimetics with and without the regulator phosphatidic acid. Biochemistry. 51(24):p.4909-21.
22. Kohri, K., et al., Biomolecular mechanism of urinary stone formation involving osteopontin. Urol Res.40(6):p.623-37.
23. Rampoldi, L., et al., The rediscovery of uromodulin (Tamm-Horsfall protein):from tubulointerstitial nephropathy to chronic kidney disease. Kidney Int.80(4):p.338-47.
24. Yamachika, T., et al., Intestinal trefoil factor:a marker of poor prognosis in gastric carcinoma. Clin Cancer Res,2002.8(5):p.1092-9.
25. Williams, R., et al., pS2 transfection of murine adenocarcinoma cell line 410.4 enhances dispersed growth pattern in a 3-D collagen gel. J Cell Sci,1996.109(Pt 1):p.63-71.
26. Thongboonkerd, V., et al., Urinary trefoil factor 1 is a novel potent inhibitor of calcium oxalate crystal growth and aggregation. J Urol,2008.179(4):p.1615-9.
27. Chutipongtanate, S., et al., Identification of human urinary trefoil factor 1 as a novel calcium oxalate crystal growth inhibitor. J Clin Invest,2005.115(12):p.3613-22.
28. Low, R.K. and M.L. Stoller, Endoscopic mapping of renal papillae for Randall's plaques in patients with urinary stone disease. J Urol,1997.158(6):p.2062-4.
29. Miller, N.L., A.P. Evan, and J.E. Lingeman, Pathogenesis of renal calculi. Urol Clin North Am,2007.34(3):p.295-313.
30. Albayrak, A., et al., The biochemical and histopathological investigation of amlodipine in ethylene glycol-induced urolithiasis rat model. Ren Fail.35(1):p.126-31.
31. Knoll, T., et al., The influence of oxalate on renal epithelial and interstitial cells. Urol Res, 2004.32(4):p.304-9.
32. McK.ee, M.D., A. Nanci, and S.R. Khan, Ultrastructural immunodetection of osteopontin and osteocalcin as major matrix components of renal calculi. J Bone Miner Res,1995. 10(12):p.1913-29.
33. Chaturvedi, L.S., et al., Oxalate selectively activates p38 mitogen-activated protein kinase and c-Jun N-terminal kinase signal transduction pathways in renal epithelial cells. J Biol Chem,2002.277(15):p.13321-30.
34.刘永旺,乙二醇诱导的草酸钙结石犬血液中抗氧化指标的变化研究,in 中国畜牧兽医学会小动物医学分会第六次学术研讨会暨中国畜牧兽医学会兽医外科学分会第十八次学术研讨会论文集.
35. Weise, A. and N. Dunker, High trefoil factor 1 (TFF1) expression in human retinoblastoma cells correlates with low growth kinetics, increased cyclin-dependent kinase (CDK) inhibitor levels and a selective down-regulation of CDK6. Histochem Cell Biol.139(2):p.323-38.
36.张励鸣,人尿液三叶因子1(TFF1)与尿路结石的相关性研究.2009.
1. Boyce, W.H. and N.M. Sulkin, Biocolloids of urine in health and in calculous disease. Ⅲ. The mucoprotein matrix of urinary calculi. J Clin Invest,1956.35(10):p.1067-79.
2. Canales, B.K., et al., Proteome of human calcium kidney stones. Urology,2010.76(4):p.1017 e13-20.
3. 杜廷义,张勇,and张云,三叶因子：从实验室研究到临床医学.动物学研究.31(1)：p.11.
4. Masiakowski, P., et al., Cloning of cDNA sequences of hormone-regulated genes from the MCF-7 human breast cancer cell line. Nucleic Acids Res,1982.10(24):p.7895-903.
5. Terada, T., et al., Characterization of the mouse TFF1 (pS2) gene promoter region. Biol Pharm Bull,2001.24(2):p.135-9.
6. Thongboonkerd, V., Proteomics and kidney stone disease. Contrib Nephrol,2008.160:p. 142-58.
7. Kraus, S., et al., MUC1 mucin and trefoil factor I protein expression in renal cell carcinoma: correlation with prognosis. Hum Pathol,2002.33(1):p.60-7.
8. 蔡华芳,学酸钙肾结石模型及其应用.中国中西医结合肾病杂志.11(8).
9. 赵宗江et al.,筛选乙二醇法复制大鼠肾结石模型最佳剂量的比较研究.中国中西医结合肾病杂志,2007.8(11)：p.3.
10. 厉建,李文峰,and 张士青,尿草酸对肾草钙结石形成和肾组织自由基影响的研究.中国实验诊断学,2008.12(7)：p.3.
11. 钟玲,et al.,乙二醇法复制肾结石模型给药途径和方法的比较.中国病理生理理杂志,2000.16(4)：p.2.
12. Hamamoto, S., K. Taguchi, and Y. Fujii, [Molecular mechanism of renal stone formation]. Clin Calcium.21(10):p.1481-7.
13. Liang, L., et al., Expression profiling of crystal-induced injury in human kidney epithelial cells. Nephron Physiol,2006.103(1):p. p53-62.
14. Khan, S.R., Role of renal epithelial cells in the initiation of calcium oxalate stones. Nephron Exp Nephrol,2004.98(2):p. e55-60.
15. Khand, F.D., et al., Mitochondrial superoxide production during oxalate-mediated oxidative stress in renal epithelial cells. Free Radic Biol Med,2002.32(12):p.1339-50.
16. Gokhale, J.A., M.D. McKee, and S.R. Khan, Immunocytochemical localization of Tamm-Horsfall protein in the kidneys of normal and nephrolithic rats. Urol Res,1996.24(4): p.201-9.
17. Sejdiu, I. and O. Torffvit, Decreased urinary concentration of Tamm-Horsfall protein is associated with development of renal failure and cardiovascular death within 20 years in type I but not in type 2 diabetic patients. Scand J Urol Nephrol,2008.42(2):p.168-74.
18. Chakraborty, J., A. A. Below, and D. Solaiman, Tamm-Horsfall protein in patients with kidney damage and diabetes. Urol Res,2004.32(2):p.79-83.
19. Hess, B., L. Zipperle, and P. Jaeger, Citrate and calcium effects on Tamm-Horsfall glycoprotein as a modifier of calcium oxalate crystal aggregation. Am J Physiol,1993.265(6 Pt 2):p. F784-91.
20. Rodriguez Camargo, D.C., N.M. Link, and S.A. Dames, The FKBP-rapamycin binding domain of human TOR undergoes strong conformational changes in the presence of membrane mimetics with and without the regulator phosphatidic acid. Biochemistry.51(24): p.4909-21.
21. Kohri, K., et al., Biomolecular mechanism of urinary stone formation involving osteopontin. Urol Res.40(6):p.623-37.
22. Rampoldi, L., et al., The rediscovery of uromodulin (Tamm-Horsfall protein):from tubulointerstitial nephropathy to chronic kidney disease. Kidney Int.80(4):p.338-47.
23. Yamachika, T., et al., Intestinal trefoil factor:a marker of poor prognosis in gastric carcinoma. Clin Cancer Res,2002.8(5):p.1092-9.
24. Williams, R., et al., pS2 transfection of murine adenocarcinoma cell line 410.4 enhances dispersed growth pattern in α 3-D collagen gel. J Cell Sci,1996.109(Pt 1):p.63-71.
25. Thongboonkerd, V., et al., Urinary trefoil factor 1 is a novel potent inhibitor of calcium oxalate crystal growth and aggregation. J Urol,2008.179(4):p.1615-9.
26. Chutipongtanate, S., et al., Identification of human urinary trefoil factor 1 as a novel calcium oxalate crystal growth inhibitor. J Clin Invest,2005.115(12):p.3613-22.
27. Low, R.K. and M.L. Stoller. Endoscopic mapping of renal papillae for Randall's plaques in patients with urinary stone disease. J Urol,1997.158(6):p.2062-4.
28. Miller, N.L., A.P. Evan, and J.E. Lingeman, Pathogenesis of renal calculi. Urol Clin North Am, 2007. 34(3): p. 295-313.
29. Albayrak, A., et al., The biochemical and histopathological investigation of amlodipine in ethylene glycol-induced urolithiasis rat model. Ren Fail. 35(1): p. 126-31.
30. Knoll, T., et al., The influence of oxalate on renal epithelial and interstitial cells. Urol Res, 2004. 32(4): p. 304-9.
31. McKee, M.D., A. Nanci, and S.R. Khan, Ultrastructural immunodetection of osteopontin and osteocalcin as major matrix components of renal calculi. J Bone Miner Res, 1995. 10(12): p. 1913-29.
32. Chaturvedi, L.S., et al., Oxalate selectively activates p38 mitogen-activated protein kinase and c-Jun N-terminal kinase signal transduction pathways in renal epithelial cells. J Biol Chem, 2002. 277(15): p. 13321-30.
33. 六永旺,乙二醇诱导的草酸钙结石走血液中抗氧化指标的变化研究,in 中国畜牧兽医学 会小动物医学分会第六次学术研讨会暨中国畜牧兽医学会兽医外科学分会第十八次学 术研讨会论文集.
34. Weise, A. and N. Dunker, High trefoil factor 1 (TFF1) expression in human retinoblastoma cells correlates with low growth kinetics, increased cyclin-dependent kinase (CDK) inhibitor levels and a selective down-regulation of CDK6. Histochem Cell Biol. 139(2): p. 323-38.
35. 张励鸣,人尿液三叶因子1TFF1)与尿路结石的相关性研究.2009.
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2. Boyce, W.H. and N.M. Sulkin, Biocolloids of urine in health and in calculous disease. Ⅲ. The mucoprotein matrix of urinary calculi. J Clin Invest,1956.35(10):p.1067-79.
3. Canales, B.K., et al., Proteome of human calcium kidney stones. Urology,2010.76(4):p.1017 e13-20.
4. 杜廷义.张勇,and张云,三叶因子：从实验室研究到临床医学.动物学研究.31(1)：p.11.
5. Masiakowski, P., et al., Cloning of cDNA sequences of hormone-regulated genes from the MCF-7 human breast cancer cell line. Nucleic Acids Res,1982.10(24):p.7895-903.
6. Terada, T., et al., Characterization of the mouse TFF1 (pS2) gene promoter region. Biol Pharm Bull,2001.24(2):p.135-9.
7. Thongboonkerd, V., Proteomics and kidney stone disease. Contrib Nephrol,2008.160:p. 142-58.
8. Kraus, S., et al., MUC1 mucin and trefoil factor 1 protein expression in renal cell carcinoma:correlation with prognosis. Hum Pathol,2002.33(1):p.60-7.
9. 蔡华芳,草酸钙肾结石模型及其应用中国中西医结合肾病杂志.11(8).
10. 赵宗江,et al.,筛选乙二醇法复制大鼠肾结石模型最佳剂量的比较研究.中国中西医结合肾病杂志,2007.8(11)：p.3.
11.厉建.李文峰.and 张士青.尿草酸对肾草钙结石形成和肾组织自由基影响的研究.中国实验诊断学,2008.12(7)：p.3.
12.钟玲.et.al.乙二醇法复制肾结石模型给药途径和方法的比较.中国病理生理杂志,2000.16(4)：p.2.
13. Hamamoto, S., K. Taguchi, and Y. Fujii, [Molecular mechanism of renal stone formation]. Clin Calcium.21(10):p.1481-7.
14. Liang, L., et al., Expression profiling of crystal-induced injury in human kidney epithelial cells. Nephron Physiol,2006.103(1):p. p53-62.
15. Khan, S.R., Role of renal epithelial cells in the initiation of calcium oxalate stones. Nephron Exp Nephrol,2004.98(2):p. e55-60.
16. Khand, F.D., et al., Mitochondrial superoxide production during oxalate-mediated oxidative stress in renal epithelial cells. Free Radic Biol Med,2002.32(12):p.1339-50.
17. Gokhale, J.A., M.D. McKee, and S.R. Khan, Immunocytochemical localization of Tamm-Horsfall protein in the kidneys of normal and nephrolithic rats. Urol Res,1996. 24(4):p.201-9.
18. Sejdiu, I. and O. Torffvit, Decreased urinary concentration of Tamm-Horsfall protein is associated with development of renal failure and cardiovascular death within 20 years in type 1 but not in type 2 diabetic patients. Scand J Urol Nephrol,2008.42(2):p.168-74.
19. Chakraborty, J., A.A. Below, and D. Solaiman, Tamm-Horsfall protein in patients with kidney damage and diabetes. Urol Res,2004.32(2):p.79-83.
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34.刘永旺,乙二醇诱导的草酸钙结石犬血液中抗氧化指标的变化研究,in 中国畜牧兽医学会小动物医学分会第六次学术研讨会暨中国畜牧兽医学会兽医外科学分会第十八次学术研讨会论文集.
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