OBJECTIVE: To observe the localization of adrenomedullin (AM) in rat kidney tissue and its inhibitory effect on the growth of cultured rat mesangial cells (MsC). METHODS: A monoclonal antibody against AM developed by ...OBJECTIVE: To observe the localization of adrenomedullin (AM) in rat kidney tissue and its inhibitory effect on the growth of cultured rat mesangial cells (MsC). METHODS: A monoclonal antibody against AM developed by our laboratory was used to detect the localization of AM protein in rat kidney tissue by avidin-biotin complex immunohistochemistry. The expressions of AM and its receptor CRLR mRNA on cultured glomerular epithelial cells (GEC) and MsC were investigated by Northern blot assay, and the possible effect of AM secreted by GEC on MsC proliferation was observed using [3H]thymidine incorporation as an index. RESULTS: A specific monoclonal antibody against AM was succesfully developed. AM was immunohistochemically localized mainly in glomeruli (GEC and endothelial cells), some cortical proximal tubules, medullary collecting duct cells, interstitial cells, vascular smooth muscle cells and endothelial cells. Northern blot assay showed that AM mRNA was expressed only on cultured GEC, but not on MsC, however, AM receptor CRLR mRNA was only expressed on MsC. GEC conditioned medium containing AM can inhibit MsC growth and AM receptor blocker CGRP8-37 may partially decreased this inhibitory effect. CONCLUSION: AM produced by GEC inhibits the proliferation of MsC, which suggests that AM as an important regulator is involved in glomerular normal physiological functions and pathologic processes.展开更多
Objective To study the expressions of MMP-2 and TIMP-2 mRNA on cultured rat mesangial cells (MsC) and in human diseased glomeruli, and to explore their significance in the development of glomerulosclerosis. Methods Th...Objective To study the expressions of MMP-2 and TIMP-2 mRNA on cultured rat mesangial cells (MsC) and in human diseased glomeruli, and to explore their significance in the development of glomerulosclerosis. Methods The expressions of MMP-2, TIMP-2, and Col ⅣmRNA on cultured rat MsC stimulated by IL-1 or/and TGF-β1were investigated through Northern blot analysis. The levels of MMP-2 and TIMP-2 mRNA expressions and immunoreacti-vity of PCNA and Col Ⅳin human diseased glomeruli from renal biopsies of lupus nephritis (LN) patients were examined by insituhybridization and immunohistochemistry, respectively. Results The levels of MMP-2, TIMP-2, and Col ⅣmRNA expressions were markedly increased on cultured rat MsC stimulated by IL-1 or/and TGF-β1. Meanwhile, upregulation of MMP-2 and TIMP-2 mRNA expressions was confirmed in diseased glomeruli from patients with various subtypes of LN, and was closely related to the positive cell number of PCNA presentation and deposition of Col Ⅳin glomeruli. Conclusion The results suggest that the over-expressions of MMP-2 and TIMP-2 mRNA on glomerular cells might play a critical role in the development of glomerulosclerosis.展开更多
Immunoglobulin A （IgA） nephropathy is one of the most common glomerulonephritis and its frequency is probably underestimated because in most patients the disease has an indolent course and the kidney biopsy is essen...Immunoglobulin A （IgA） nephropathy is one of the most common glomerulonephritis and its frequency is probably underestimated because in most patients the disease has an indolent course and the kidney biopsy is essential for the diagnosis. In the last years its pathogenesis has been better identifed even if still now several questions remain to be answered. The genetic wide association studies have allowed to identifying the relevance of genetics and several putative genes have been identified. The genetics has also allowed explaining why some ancestral groups are affected with higher frequency. To date is clear that IgA nephropathy is related to auto antibodies against immunoglobulin A1 （IgA1） with poor O-glycosylation. The role of mucosal infections is confirmed, but which are the pathogens involved and which is the role of Toll-like receptor polymorphism is less clear. Similarly to date whether the disease is due to the circulating immunocomplexes deposition on the mesangium or whether the antigen is already present on the mesangial cell as a “lanthanic” deposition remains to be clarifed. Finally also the link between the mesangial and the podocyte injury and the tubulointerstitial scarring, as well as the mechanisms involved need to be better clarifed.展开更多
Objective: To investigate the role of laminins in the pathogensis of mesangial pro-liferative glomeruonephritis (MsPGN) in children. Methods: Eighteen renal biopsy specimens of MsPGN and 6 normal kidneys were studied ...Objective: To investigate the role of laminins in the pathogensis of mesangial pro-liferative glomeruonephritis (MsPGN) in children. Methods: Eighteen renal biopsy specimens of MsPGN and 6 normal kidneys were studied by means of immunohistochemistry and in situ hybridization. Results: ① Protein of α1 chain and γ1 chain of laminin increased around the segments of prolifera-tive mesangium. Increased expression of α2 and β1 proteins was found in the segments with mesangial proliferation whereas the β2 chain expression decreased in these areas. ② The mRNA expression of α1, α2, β1 and γ1 increased to different degrees in glomeruli with mesangial proliferation. But no difference was detected among Mild, Moderate, and Severe MsPGN. Conclusion:① The quantitative and qualitative alterations of laminin chains' distribution were found in the measngial proliferative glomeruli. The proliferative mesangial cells were the origins of abnormal accumulation and expression of laminins. ② These changes may be the basis of the progresses of MsPGN.展开更多
To examine the effect of multi-glycoside of Tripterygium wilfordii Hook. f. （GTW） on proteinuria and acute glomerular immune lesion in experimental mesangial proliferative glomerulonephritis （MsPGN） induced by ant...To examine the effect of multi-glycoside of Tripterygium wilfordii Hook. f. （GTW） on proteinuria and acute glomerular immune lesion in experimental mesangial proliferative glomerulonephritis （MsPGN） induced by anti-Thyl. 1 monoelonal antibody （mAb） 1-22-3. The reversible model of MsPGN with anti-Thyl. 1 mAb 1-22-3 was established. After 7 days of oral treatment with GTW （ 100 mg/kg per day） and vehicle （distilled water, 5 ml/kg per day）, its effects on proteinuria, renal functions, mesangial morphological change, glomerular macrophage accumulation, and mRNA expressions of cytokines （PDGF-BB and MCP-1 ） were evaluated by light microscope （LM）, immunofluorescence （IF）, and reverse transcription polymerase chain reaction （RT-PCR）. It was found that GTW ameliorated proteinuria （on day 3 and day 7）, mesangial proliferation （total cell number, matrix expansion, a-smooth muscle actin expression, and collagen type Ⅰ expression） and macrophage accumulation （ED3^＋ ） in experimental MsPGN. In addition, GTW significantly suppressed the increased mRNA expressions for MCP-1 （67.6% to eontrol group, P 〈 0.01） together with the tendency to reduce the expression of PDGF （24.44% to control group） on day 7. It is concluded that GTW can not only decrease proteinuria, but also ameliorate acute mesangial alterations and glomerular activated macrophage accumulation probably by reduction of cytokines. These data indicate that GTW is an effective agent for early MsPGN.展开更多
Objective To investigate the effect of immune cell from idiopathic nephrotic children on extracellular matrix (ECM) synthesis by cultured rat glomerular epithelial cell (GEC) and on the proliferation of mesangial cel...Objective To investigate the effect of immune cell from idiopathic nephrotic children on extracellular matrix (ECM) synthesis by cultured rat glomerular epithelial cell (GEC) and on the proliferation of mesangial cell (GMC). Methods Twenty-eight children with idiopathic nephrotic syndrome and 15 age-matched healthy children were randomly selected and divided into 4 groups: Group 1, untreated nephrotic children; Group 2, glucocorticoid treated nephrotic children; Group 3, children undergoing glucocorticoid treatment with negative proteinuria; and Group 4, normal control. The peripheral blood mononuclear cells (PBMC) were collected from these children and PBMC conditioned medium (PBMC-CM) were prepared. The PBMC-CM was co-cultured with GEC and GMC respectively. The concentrations of collagen, laminin, collagen Ⅲ and collagen Ⅳ in the GEC and PBMC-CM co-culture medium were investigated. The GMC proliferation was measured by the 3 H-thymidin incorporation method. Results The 3 H-proline incorporation coefficients of the GEC treated with the PBMC-CM of the 4 groups were 0.93, 1.24, 1.23, and 1.11, respectively. The laminin inhibitory coefficients of the 4 groups were 0.95, 1.02, 1.01, and 1.04, respectively. The inhibitory coefficients of collagen Ⅲ were 0.97, 1.00, 0.99, and 1.01, respectively, for the 4 groups. All these parameters showed a significant difference between Group 1 and the other 3 groups (P<0.05). However, there was no significant difference in the inhibitory coefficient of collagen Ⅳ between each two of the 4 groups (1.04, 1.05, 1.04, 1.08, P>0.05). The 3 H-thymidine incorporation coefficients of GMC responsive to PBMC-CM were 1.21, 1.53, 1.50, and 1.10, respectively, and no significant difference was found between the 4 groups (P>0.05). Conclusion The results suggested that the circulating immune cells from idiopathic nephrotic children have a direct effect on some ECM component synthesis in cultured rat GEC; the bio-activity of immune cells could be neutralized by administering glucocorticoid; and the circulating immune cells of nephrotic children have no direct effect on GMC proliferation.展开更多
文摘双硫仑作为一种治疗慢性酒精中毒的药物在临床中广泛使用。近年来,研究者提出了双硫仑治疗癌症的具体机制,如抑制乙醛脱氢酶(acetaldehyde dehydrogenase,ALDH)的活性、提高细胞内活性氧(reactive oxygen species,ROS)的浓度、抑制核因子kappa-B(nuclear factor kappa-B,NF-κB)的活性,促进与核蛋白定位蛋白4(nuclear protein localization protein 4,NPL4)的结合、抑制FROUNT蛋白等,并在多种癌症模型中证明了双硫仑的抗癌活性。抗肾小球基底膜型肾小球肾炎是急进性肾小球肾炎中的一种类型,该病一旦被确诊,就需要第一时间给予治疗,尽量帮助患者缓解症状、改善预后。研究表明,双硫仑可通过抑制C-C趋化因子受体2型/C-C趋化因子受体5型(C-C chemokine receptor type 2/C-C chemokine receptor type 5,CCR-2/CCR-5)和FROUNT蛋白之间的相互作用来抑制巨噬细胞的迁移、聚集、活化来缓解抗肾小球基底膜型肾小球肾炎,这表明双硫仑对该类患者具有潜在的治疗价值。本文简要回顾了双硫仑最新研究中阐明的相关作用分子机制,展望了未来双硫仑作为新的药物治疗抗肾小球基底膜型肾小球肾炎的前景。
文摘OBJECTIVE: To observe the localization of adrenomedullin (AM) in rat kidney tissue and its inhibitory effect on the growth of cultured rat mesangial cells (MsC). METHODS: A monoclonal antibody against AM developed by our laboratory was used to detect the localization of AM protein in rat kidney tissue by avidin-biotin complex immunohistochemistry. The expressions of AM and its receptor CRLR mRNA on cultured glomerular epithelial cells (GEC) and MsC were investigated by Northern blot assay, and the possible effect of AM secreted by GEC on MsC proliferation was observed using [3H]thymidine incorporation as an index. RESULTS: A specific monoclonal antibody against AM was succesfully developed. AM was immunohistochemically localized mainly in glomeruli (GEC and endothelial cells), some cortical proximal tubules, medullary collecting duct cells, interstitial cells, vascular smooth muscle cells and endothelial cells. Northern blot assay showed that AM mRNA was expressed only on cultured GEC, but not on MsC, however, AM receptor CRLR mRNA was only expressed on MsC. GEC conditioned medium containing AM can inhibit MsC growth and AM receptor blocker CGRP8-37 may partially decreased this inhibitory effect. CONCLUSION: AM produced by GEC inhibits the proliferation of MsC, which suggests that AM as an important regulator is involved in glomerular normal physiological functions and pathologic processes.
基金Supported by Shanghai Municipal Science and Technology Develop-ment Funds (01JC14018).
文摘Objective To study the expressions of MMP-2 and TIMP-2 mRNA on cultured rat mesangial cells (MsC) and in human diseased glomeruli, and to explore their significance in the development of glomerulosclerosis. Methods The expressions of MMP-2, TIMP-2, and Col ⅣmRNA on cultured rat MsC stimulated by IL-1 or/and TGF-β1were investigated through Northern blot analysis. The levels of MMP-2 and TIMP-2 mRNA expressions and immunoreacti-vity of PCNA and Col Ⅳin human diseased glomeruli from renal biopsies of lupus nephritis (LN) patients were examined by insituhybridization and immunohistochemistry, respectively. Results The levels of MMP-2, TIMP-2, and Col ⅣmRNA expressions were markedly increased on cultured rat MsC stimulated by IL-1 or/and TGF-β1. Meanwhile, upregulation of MMP-2 and TIMP-2 mRNA expressions was confirmed in diseased glomeruli from patients with various subtypes of LN, and was closely related to the positive cell number of PCNA presentation and deposition of Col Ⅳin glomeruli. Conclusion The results suggest that the over-expressions of MMP-2 and TIMP-2 mRNA on glomerular cells might play a critical role in the development of glomerulosclerosis.
文摘Immunoglobulin A （IgA） nephropathy is one of the most common glomerulonephritis and its frequency is probably underestimated because in most patients the disease has an indolent course and the kidney biopsy is essential for the diagnosis. In the last years its pathogenesis has been better identifed even if still now several questions remain to be answered. The genetic wide association studies have allowed to identifying the relevance of genetics and several putative genes have been identified. The genetics has also allowed explaining why some ancestral groups are affected with higher frequency. To date is clear that IgA nephropathy is related to auto antibodies against immunoglobulin A1 （IgA1） with poor O-glycosylation. The role of mucosal infections is confirmed, but which are the pathogens involved and which is the role of Toll-like receptor polymorphism is less clear. Similarly to date whether the disease is due to the circulating immunocomplexes deposition on the mesangium or whether the antigen is already present on the mesangial cell as a “lanthanic” deposition remains to be clarifed. Finally also the link between the mesangial and the podocyte injury and the tubulointerstitial scarring, as well as the mechanisms involved need to be better clarifed.
基金Supported by Grant from the Natural Science Foundation of Education Committee of Jiangsu Province(96047)
文摘Objective: To investigate the role of laminins in the pathogensis of mesangial pro-liferative glomeruonephritis (MsPGN) in children. Methods: Eighteen renal biopsy specimens of MsPGN and 6 normal kidneys were studied by means of immunohistochemistry and in situ hybridization. Results: ① Protein of α1 chain and γ1 chain of laminin increased around the segments of prolifera-tive mesangium. Increased expression of α2 and β1 proteins was found in the segments with mesangial proliferation whereas the β2 chain expression decreased in these areas. ② The mRNA expression of α1, α2, β1 and γ1 increased to different degrees in glomeruli with mesangial proliferation. But no difference was detected among Mild, Moderate, and Severe MsPGN. Conclusion:① The quantitative and qualitative alterations of laminin chains' distribution were found in the measngial proliferative glomeruli. The proliferative mesangial cells were the origins of abnormal accumulation and expression of laminins. ② These changes may be the basis of the progresses of MsPGN.
文摘To examine the effect of multi-glycoside of Tripterygium wilfordii Hook. f. （GTW） on proteinuria and acute glomerular immune lesion in experimental mesangial proliferative glomerulonephritis （MsPGN） induced by anti-Thyl. 1 monoelonal antibody （mAb） 1-22-3. The reversible model of MsPGN with anti-Thyl. 1 mAb 1-22-3 was established. After 7 days of oral treatment with GTW （ 100 mg/kg per day） and vehicle （distilled water, 5 ml/kg per day）, its effects on proteinuria, renal functions, mesangial morphological change, glomerular macrophage accumulation, and mRNA expressions of cytokines （PDGF-BB and MCP-1 ） were evaluated by light microscope （LM）, immunofluorescence （IF）, and reverse transcription polymerase chain reaction （RT-PCR）. It was found that GTW ameliorated proteinuria （on day 3 and day 7）, mesangial proliferation （total cell number, matrix expansion, a-smooth muscle actin expression, and collagen type Ⅰ expression） and macrophage accumulation （ED3^＋ ） in experimental MsPGN. In addition, GTW significantly suppressed the increased mRNA expressions for MCP-1 （67.6% to eontrol group, P 〈 0.01） together with the tendency to reduce the expression of PDGF （24.44% to control group） on day 7. It is concluded that GTW can not only decrease proteinuria, but also ameliorate acute mesangial alterations and glomerular activated macrophage accumulation probably by reduction of cytokines. These data indicate that GTW is an effective agent for early MsPGN.
基金ThisworkwasgrantedfromtheMedicalScienceandTechnicalFoundationofthePLA (No 96D0 36) .
文摘Objective To investigate the effect of immune cell from idiopathic nephrotic children on extracellular matrix (ECM) synthesis by cultured rat glomerular epithelial cell (GEC) and on the proliferation of mesangial cell (GMC). Methods Twenty-eight children with idiopathic nephrotic syndrome and 15 age-matched healthy children were randomly selected and divided into 4 groups: Group 1, untreated nephrotic children; Group 2, glucocorticoid treated nephrotic children; Group 3, children undergoing glucocorticoid treatment with negative proteinuria; and Group 4, normal control. The peripheral blood mononuclear cells (PBMC) were collected from these children and PBMC conditioned medium (PBMC-CM) were prepared. The PBMC-CM was co-cultured with GEC and GMC respectively. The concentrations of collagen, laminin, collagen Ⅲ and collagen Ⅳ in the GEC and PBMC-CM co-culture medium were investigated. The GMC proliferation was measured by the 3 H-thymidin incorporation method. Results The 3 H-proline incorporation coefficients of the GEC treated with the PBMC-CM of the 4 groups were 0.93, 1.24, 1.23, and 1.11, respectively. The laminin inhibitory coefficients of the 4 groups were 0.95, 1.02, 1.01, and 1.04, respectively. The inhibitory coefficients of collagen Ⅲ were 0.97, 1.00, 0.99, and 1.01, respectively, for the 4 groups. All these parameters showed a significant difference between Group 1 and the other 3 groups (P<0.05). However, there was no significant difference in the inhibitory coefficient of collagen Ⅳ between each two of the 4 groups (1.04, 1.05, 1.04, 1.08, P>0.05). The 3 H-thymidine incorporation coefficients of GMC responsive to PBMC-CM were 1.21, 1.53, 1.50, and 1.10, respectively, and no significant difference was found between the 4 groups (P>0.05). Conclusion The results suggested that the circulating immune cells from idiopathic nephrotic children have a direct effect on some ECM component synthesis in cultured rat GEC; the bio-activity of immune cells could be neutralized by administering glucocorticoid; and the circulating immune cells of nephrotic children have no direct effect on GMC proliferation.
