Dissertation > Excellent graduate degree dissertation topics show

Effects of PTKs and Genistein on Human Renal Tubular Epithelial-Myofibroblast Transdifferention in Vitro

Author: MeiZuoMing
Tutor: Zhang
School: Third Military Medical University
Course: Internal Medicine
Keywords: epithelial-myofibroblast transdifferentiation (EMT) acid fibroblast growth factor (aFGF) α-smooth muscle actin (α-SMA) human renal tubular epithelial cell(HKC) genistein protien tyrosine kinases(PTKs) transforming growth factor β1(TGFβ1)
CLC: R692.9
Type: Master's thesis
Year: 2002
Downloads: 51
Quote: 1
Read: Download Dissertation

Abstract


Objective:Interstitial fibrosis has shown a strong correlation with the progression of kidney disease to renal insufficiency.This proccess is associated at least in some forms of interstitial fibrosis with a differentiation of tubular epithelial cells into myofibroblasts.However,the mechanism underlying this differentiation of tubular epithelial cells is poorly understood.This study investigated the possible roles of protein tyrosine kinases(PTKs) in the phenotypic transformation of cultured human renal tubular epithelial cell line(HKC).Methods: The HKC cells were divided into three groups:(1)serum-free (negative control);(2)aFGF(positve control);(3)aFGF-inhibtion(treated with aFGF + genistein).Cells were cultured for 72h on collagen typeⅠ-coated plate.HKC cells were analyzed by immunohitochemistry with monoantibodies(mAbs) to α-smooth muscle actin(α-SMA),vimentin and cytokeratin, respectively.Morphlogy was determined using phase contrast microscopy and electron microscope.The number of HKC cells stained with the anti-α-SMA mAb was determined by counting the number of positive-stained cells and data are expressed as the mean percentage.Cell motility was estimated by the ability of cells located at the edge of an artificial "Scarification ".The expression of FN in the supernatant of HKC cells was assessed with ELISA. Flow cytometry was adopted to measure the activity of PTKs. TGFβ1 mRNA expression was examined by revers transcription- polymerase chain reaction(RT-PCR) after cells were allowed to grow for 24h<WP=5>in the presence or absence of various factors and reagents.Results: HKC cells in serum control showed a classic epithelial morphology.In contrast,culture in 80ng/ml aFGF caused profound changes in morphology.These involved hypertrophy,a loss of apical-basal polarity, identifiable junctional complexes and microvilli,with cells becoming elongated and invasive, and formation of a new front-end back-end polarity, and rich in ribosome, rough endoplasmic reticulum and Golgi body.These morphological changes were companied by phenotypic changes.Immunohistochemistry staining showed that the addition of aFGF to subconfluent cell culture caused a loss of the epithelial marker (cytokeratin)and increased expression of mesenchymal marker (vimentin and α-SMA).However,medium wih aFGF +genistein,these expressions were lower than those in positive controls.The percentage of α-SMA +HKC cells in the positive controls was significantly higher than that in negative controls(60.60±2.70% vs 3.35±1.45%,P<0.01); While the percentage of α-SMA +HKC cells in aFGF +genistein group at genistein dose of 48,96μg/ml(3.90±2.09%,3.98±1.75%) was dramtically lower than that in positive controls(P<0.01).The content of FN in the supernatant in aFGF +genistein group at genistein dose of 24,48μg/ml was lower than that in positive controls(0.571±0.016, 0.499±0.002 vs 0.701±0.047,P<0.05) and FN in positive control was markedly higher than that in negative controls(0.701±0.047 vs 0.498±0.004,P<0.01).There was a dose-dependent increase in the level of cells expressing TGFβ1 mRNA with increasing concentrations of aFGF.The level of TGFβ1 mRNA in cells by the addition of aFGF(40,80 ng/ml) was much higher than negative control(0.583±0.015,0.947±0.015 vs 0.383±0.015,P<0.01);However, the level of TGFβ1 mRNA in aFGF +genistein group at genistein dose of 24,48μg/ml was markedly lower than that in positive controls(0.617±0.025, 0.400±0.026 vs 0.947±0.015,P<0.01).In the course of epithelial cell transdifferent to<WP=6>myofibroblast,PTKs was activated in a aFGF dose-denpendent manner..This represented 1.16,1.17,3.46times increase in the activity of PTKs (aFGF20,40,80ng/ml vs negative controls).On the contrary,inhibition of the activity of PTKs was achieved in aFGF +genistein group at genistein dose of 6,12,24,48μg/ml.(a 45.6%,53.5%,66%,81.9% decrease of PTKs of positive group respectively.)Conclusion:1. The transdifferentiaton of HKC cells can be induced by aFGF .The present model may serve as a useful tool for

Related Dissertations

  1. The Research of Red Clover Isoflavones Transforming into Genistein and Daidzein,TQ464
  2. The Investigation of the Protective Effects of 7-difluoromethyl-5,4’-dimethoxygenistein on Atherosclerosis in the Rabbit Model,R285.5
  3. The Experimental Study of Soybean Extraction on Benign Prostatic Hyperplasia,R285.5
  4. Studies on Tablet of Semen Sojae Preparatrm Isoflavone Preparation,TQ461
  5. Experimental Study on the Effects of GDNF on Perineural Invasion of Pancreatic Carcinoma,R735.9
  6. Effect of Genistein on Growth and Apoptosis of Colon Cancer LOVO Cells and Its Mechanisms,R735.35
  7. Study of the Combined Effect and Its Mechanism of Genistein and EGF on Human Breast Cancer Cell MCF-7,R737.9
  8. The Oxidative Stress on RPE Cells and Mechanism of Genistein in Hypoxia,R774.1
  9. Isoflavones Regulate Target Genes Transcription Through Estrogen Receptors,R285
  10. The Study of the Effect of Genistein on the Proliferation and Mechanism of Hepatoma Cell Line SMMC-7721 Cells,R735.7
  11. Genistein Suppresses in Vitro Invasion and Metastasis of Human Hepatoma Cells SMMC-7721,R735.7
  12. Inhibitory Effect of Genistein on Human Ovarian Cancer Cell Line HO-8910PM in Nude Mice,R737.31
  13. Effects of Genistein and Diethylstilbestrol on Bone Metabolism in Orchidectomized Rats,R580
  14. The Effect of Genistein on Tumor Recurrence and Metastasis after Partial Hepatectomy in Liver Cancer,R735.7
  15. Synthesis and Recognition Mechanism of Molecularly Imprinted Polymers for Phenylurea Herbicides and Genistein and Their Application Study,TQ450.1
  16. Study on Effects of Genistein on Diabetic Rats and Its Effects on CD30 and other Genes’ Expression in Wnt Signal Pathway in hESCs,R285
  17. Studies on Chemical Constituents in Soybean,R284
  18. The Expression and Significance of Integrin-linked Kinase in the Renal Interstitial of Primary Glomerulonephritis,R692.3
  19. Mechanisms of Inhibiting Proliferation and Inducing Apoptosis of Human Gastric Cancer Cell Line BGC-823 by Genistein,R735.2
  20. Effect on the Proliferation of Rabbit Conjunctival Fibroblasts in Vitro by Genistein,R775
  21. Ⅰ.Electrophysiological Effects of Phytoestrogen Genistein on Spontaneous Activity of Rabbit Atrioventricular Node Cells Ⅱ.Effects of Capsaicin on Electrophysiology of Cardiomyocytes and Underlying Mechanism,Q24

CLC: > Medicine, health > Surgery > Urology ( urinary and reproductive system diseases) > Kidney disease > Other diseases
© 2012 www.DissertationTopic.Net  Mobile