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Effects of Mycophenolic Acid on High Glucose-induced Expression of TGF-β and CTGF in mesangial cells

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2006年第3期26卷 292-293,304页

作者：吕永曼陈俊英邵菊芳 Department of Nephrology Tongji Hospital Huazhong University of Science and Technology Wuhan 430030 China

The effects of mycophenolic acid （MPA） on high glucose-induced expression of transforming growth factor-β （TGF-β） and connective tissue growth factor （CTGF） in mesangial cells （MC） were investigated. Rat MC were cultured in the presence of different concentrations of MPA （1.0 and 10.0 μmol/L） or MPA plus high glucose for 72 h. The expression of TGF-β and CTGF was detected by Western blot. The results showed that high glucose could induce the expression of TGF-β and CTGF in MC, but MPA could inhibit this effects. MPA did not influence the expression of TGF-β and CTGF in normal glucose. It was concluded that MPA might prevent the progression of diabetic nephropathy by inhibiting the expression of TGF-β and CTGF in MC.

关键词： mycophenolic acid transforming growth factor-β connective tissue growth factor mesangial cell diabetic nephropathy

Effect of Astragalin on matrix secretion and β1 integrin mRNA expression in human mesangial cells

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Chinese Medical Journal 1999年第12期 7-11页

作者：倪兆慧张庆怡钱家麒王利民

Objective To investigate the effect of Astragalin on human renal mesangial cells Methods Cultured human mesangial cells were treated with Astragalin and Astragalin serum in different concentrations in the presence or absence of PDGF BB, the proliferation and type Ⅳ collagen secretion of mesangial cells were measured by MTT assay and ELISA, and expression of β1 integrin gene was estimated by reverse transcription polymerase chain reaction (RT PCR) method, sespectively Results After 72 hours Astragalin or Astragalin serum treatment, the proliferation of mesangial cells induced by PDGF BB was inhibited significantly in a dose dependent manner compared with untreated controls ( P cell proliferation and matrix over synthesis which might be mediated, at least, partly by decrease of β1 integrin gene over expression The study suggested that Astragalin might play a role in preventing the progression of chronic renal diseases

关键词： Astragalin mesangial cell β1 integrin type Ⅳ collagen

Regulation of the expression and function of glucose transporter-1 by TGF- β1 and high glucose in mesangial cells

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Chinese Medical Journal 2000年第6期113卷 28-33页

作者：章精刘志红刘浩李颖健黎磊石 Research Institute of Nephrology Jinling Hospital Nanjing University School of Medicine Nanjing 210002 China.

Objective To study the effects of high glucose and transforming growth factor β1 (TGF β1) on the expression and function of glucose transporter 1 (GLUT1) in mouse mesangial cells Methods Cultured mouse mesangial cells were used The expression of GLUT1 mRNA was detected by Northern Blot; glucose uptake and its kinetics were determined with a 2 Deoxy [ 3H] D glucose uptake assay Results mesangial cells exposed to enriched glucose medium (20?mmol/L) for 72 hours demonstrated a decrease in both GLUT1 mRNA and V max for uptake of the glucose analog, 2 deoxy D glucose (2DOG), as compared to mesangial cells cultured in physiologic glucose concentrations(5 5?mmol/L) In contrast, hypertonic mannitol had no effect on GLUT1 mRNA levels TGF β1 treatment for 10 hours stimulated 2DOG uptake, both in 5 5?mmol/L and 20?mmol/L glucose medium, by approximately 4 28 fold in a dose dependent manner (2?ng/ml maximum) Kinetic analysis of 2DOG uptake revealed an increase in V max and a decrease in K m in the presence of TGF β1 TGF β1 also up regulated the expression of GLUT1 mRNA in mesangial cells The addition of anti TGF β neutralizing antibody (30?μg/ml) in mesangial cells cultured in enriched glucose medium (20?mmol/L) led to a 40% decrease in 2DOG uptake Conclusions The expression of GLUT1 can be suppressed by exposure of mesangial cells to high glucose medium, which may serve as a protective mechanism against possible adverse effects of excessive glucose flux into cells TGF β1 stimulates glucose uptake by enhancing the expression and function of GLUT1 in mesangial cells This effect is independent of the glucose milieu in the cultured medium

关键词： glucose transporter-1 hyperglycemia TGF- β1 mesangial cell diabetic nephropathy

Effects of Xiaoke granule on transforming growth factor-beta_1 expression and proliferation in rat mesangial cells

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Chinese Medical Journal 2006年第21期119卷 1839-1842页

作者： JI Xiao-mei WANG Qian GONG Mu-xin DU Yu-qiong JIA De-xian School of Chinese Medicine Capital University of Medical Sciences Beijing 100013 China School of Preclinical Medicine Beijing University of Chinese Medicine Beijing 100029 China

Diabetic nephropathy, one of the major causes of death of diabetes patients, is diagnosed as the thickening of glomerular basement membrane and progressive expansion of the glomerular mesangium and tubulointerstitium. Intensive studies have shown that hyperglycemia is the key factor for renal sclerosis which can lead to end-stage renal disease for diabetic patients.^1,2 Our previous studies demonstrated that Xiaoke granule can inhibit the progression of diabetic nephropathy. However, its mechanisms remain unknown.^3,4 In this study, we found that Xiaoke granule coincidently depresses transforming growth factor-beta1 （TGF-β1） expression and inhibits the effect of high glucose on mesangial cell proliferation. This might suggest that the effect of Xiaoke granule on inhibiting progression of diabetic nephropathy through down-regulating TGF-β1 expression.

关键词： Chinese medicine rat mesangial cell transforming growth factor-beta1

Effects of Nephritis No.3 Recipe on Nitric Oxide,Nitric Oxide Synthase Secreted by Cultured mesangial cells in Rats and the Gene Expression of Inducible Nitric Oxide Synthase

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Chinese Journal of Integrated Traditional and Western Medicine 2003年第3期9卷 210-214页

作者：陈志强黄怀鹏黄文政朱小棣林清棋 Institute of Integrative Chinese and Western Medicine Hebei Medical UniversityShijiazhuang 050017 Nanjing University of TCM The Affiliated Hospital of Tianjin College of TCM The Affilated Hospital of Tianjin College of TCM

Objective:To explore the effect of the Nephritis No. 3 (N-3) recipe on nitric oxide (NO), nitric oxide synthase (NOS) secreted by cultured mesangial cells (MC) and its gene expression of the in-ducible nitric oxide synthase (iNOS). Methods:The drug (nephritis No. 3)-containing serum was prepared with serum pharmacological technique, and then was applied to react on mesangial cells cultured in fetal calf serum (FCS) and cells cultured in FCS plus lipopolysaccharide. To observe the secretion of NO and NOS and the gene expression of iNOS by means of RT-PCR. Results:Under the two kinds of culture conditions, the content of NO and NOS in the groups with drug-containing serum were higher than those without drug-containing serum (P<0. 05, P<0. 01), and the expression of iNOS mRNA was up-regulated too. Conclusion: The N-3 could significantly promote the secretion of NO and NOS and the mRNA expression of iNOS in rats.

关键词： Nephritis No. 3 recipe mesangial cell nitric oxide nitric oxide synthase

Jak_1/STAT_3 pathway mediates the inhibition of lipoxin A_4 on TNF-α-induced DNA synthesis of glomerular mesangial cells in rats

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Journal of Nanjing Medical University 2005年第5期19卷 223-226页

作者： Shenghua Wu Chao LU Ling Dong Ziqing Chen Department of Pediatrics the First Affiliated Hospital of Nanjing Medical University Nanjing 210029 China Central Laboratory the First Affiliated Hospital of Nanjing Medical University Nanjing 210029 China

Objective： To examine whether lipoxin A4 （LXA4） has an inhibitory effect on tumor necrosis factor-α（TNF-α-induced DNA synthesis of glomerular mesangial cells of rat, and explore the molecular mechanisms of LXA4 action. Methods： Glomerular mesangial cells of rat were cultured and preincubated with LXA4 at different concentrations, and then treated with TNF-α（ 10 ng/ml）. DNA synthesis was assessed by the incorporation of [^3H]-thymidine in mesangial cells. Expression of cyclin E protein was determined by Western blotting analysis. Activities of signal transducers and activators of transcription-3 （STAT3） were analyzed by electrophoretic mobility shift assay （EMSA）. Results： TNF-α-stimulated DNA synthesis of mesangial cells, upregulafion of cyclin E protein and STAT3 activities were inhibited by LXA4 in a dose-dependent manner. Conclusion： TNF-α-induced DNA synthesis of mesangial cells can be inhibited by TXA4 probably through the mechanism of Jak1/STAT3 pathway-dependent signal transduction.

关键词： lipoxin tumor necrosis factor DNA synthesis cyclin STAT mesangial cell

SIGNIFICANCE OF MMP-2 AND TIMP-2 mRNA EXPRESSIONS ON GLOMERULAR cellS IN THE DEVELOPMENT OF GLOMERULOSCLEROSIS

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Chinese Medical Sciences Journal 2004年第2期19卷 84-88页

作者： Zhi-gangZhang Xue-guangLiu Guang-pingChen Xiu-rongZhang Mu-yiGuo DepartmentofPathology ShanghaiMedicalCollegeofFudanUniversityShanghai200032

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.

关键词： matrix metalloproteinase tissue inhibitor of metalloproteinase mesangial cell human lupus nephritis

Shp-2／NF-κB Pathway Mediates the Inhibition of Lipoxin A4 onIL-1β-induced Synthesis of IL-6 in Glomerular mesangial cells

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Journal of Nanjing Medical University 2004年第4期18卷 167-171页

作者： WUSheng-hua LUChao DONGLing CHENZi-qing DepartmentofPediatrics CentralLaboratorytheFirstAffiliatedHospitalofNanjingMedicalUniversityNanjing210029P.R.China

Objective: To examine whether lipoxin A 4 (LXA 4) has an antagonistic effect on IL-1β-induced synthesis of IL-6 in glomerular mesangial cells, and to explore the molecular mechanisms of signal pathway in LXA 4 actions. Methods: The glomerular mesangial cells of rat were cultured and treated with IL-1β, with or without preincubation with LXA 4 at different concentrations. The amount of IL-6 in the supernatant of cells was analyzed by enzyme-linked immunosorbent assay(ELISA). The expressions of mRNA of IL-6 were determined by RT-PCR. The expressions of Src homology 2(SH 2) containing protein-tyrosine phosphatase 2(Shp-2) were assessed by immunoprecipitation and immunoblotting. Activities of DNA-binding of nuclear factor-kappa B(NF-κB) were measured by electrophoretic mobility shift assay(EMSA). Results: IL-1β-stimulated secretion of protein and expression of mRNA of IL-6 in mesangial cells were inhibited by LXA 4 in a dose-dependent manner. LXA 4 antagonizes the phosphorylation of Shp-2 and activities of NF-κB induced by IL-1β. Conclusion: LXA 4 antagonists IL-1β-induced synthesis of IL-6 in glomerular mesangial cells through the mechanism of Shp-2/NF-κB pathway-dependent signal transduction.

关键词： lipoxin interleukin nuclear factor-kappa B Shp-2 mesangial cell

Effects of Shikonin on Proliferation, Apoptosis, and Extracellular Matrix of Human mesangial cells

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Journal of Nanjing Medical University 2004年第6期8卷 304-307页

作者：李海涛李晓冬朱荃南京中医药大学药学院

Objective:To investigate the effect of shikonin on the proliferation, apoptosis and extracellular matrix (ECM) of human mesangial cells (MC). Methods: MC was cultured in vitro with different concentrations of glucose (30, 50, 80 mmol/L). The cell growth was observed by using MTT method and apoptosis by using an aunexin-V-Fluos. Immunohistochemical studies for Laminin (LN), Fibronectin (FN) and type Ⅳ Collagens (Col Ⅳ) were measured. Results: Shikonin inhibited their growth (Pcells. Shikonin 0.05 mmol/L significantly reduced the secretion of LN, FN and Col Ⅳ from MC (P<0.05) cultured in 30, 50 and 80 mmol/L glucose. Conclusion: Shikonin could prevent or treat diabetic nephropathy (DN) and glomerulosclerosis (GS).

关键词： shikonin mesangial cell proliferation extracellular matrix apoptosis glomerulosclerosis