Dissertation > Excellent graduate degree dissertation topics show

Triptolide Protects Podocytes from Interleukin-13 Induced Injury in Vitro

Author: HongYiMei
Tutor: LiuZhiHong；LiLeiShi
School: Second Military Medical University
Course: Internal Medicine
Keywords: Interleukin- 13 Proteinuria Podocytes Signal transducer - activator of transcription 6 Triptolide
CLC: R285.5
Type: Master's thesis
Year: 2009
Downloads: 92
Quote: 0
Read: Download Dissertation

Abstract


Podocytes minimal change nephropathy (MCD) and focal segmental glomerulosclerosis (FSGS) in renal tissue lesion formation damage target cells. Many studies have shown that T cell dysfunction, Th2 cells produce cytokines (IL-4, IL-13, etc.) plays an important role in the development of FSGS and MCD lesions. Another study found that serum interleukin-13 (IL-13) levels than normal rise with FSGS patients. Therefore, the direct effect of IL-13 foot cell may be one of the pathogenesis of FSGS. This study examined in vitro podocyte expression of IL-13 receptor, observe the direct damage effects of IL-13 foot cell, and to explore its mechanism of action. FSGS treatment drug of choice is still glucocorticoids, but the complete remission rate was less than 30%. Proteinuria strive to make treatment to achieve mitigation can significantly improve the prognosis of patients in a short time. Triptolide therapy for glomerular diseases for more than 30 years of history, has a positive meaning in the treatment of FSGS. Previous studies also found that the podocyte injury induced by triptolide puromycin (puromycin aminonucleoside, PA) any protective effect. Since IL-13-induced podocyte injury is one of the possible pathogenesis of FSGS, therefore, we continue to explore the protective effect of Triptolide in vitro IL-13-induced podocyte injury. The first part of the study consists of two parts: IL-13-induced podocyte injury and its mechanism of action studies Objective: podocyte MCD and FSGS kidney tissue lesions formed major damage to the target cells. Many studies have shown that T cell dysfunction, Th2 cells produce cytokines (IL-4, IL-13, etc.) plays an important role in the development of FSGS and MCD lesions. To explore the Th2 cytokine IL-13 is directly mediated podocyte injury, we use a temperature-sensitive mouse podocytes Department (heat sensitive mouse podocyte, the HSMP) to detect the IL-13 receptor expression on podocytes effect of IL- 13 pairs of direct podocyte injury, and to explore its mechanism of action. Methods: RT-PCR and immunohistochemical methods to detect the HSMP on IL-13 receptor mRNA and protein expression. Using different concentrations (10ng/ml, 50ng/ml, 100ng/ml), IL-13 intervention the podocytes time (8h, 12h, 24h), immunofluorescence analysis of the connection between the foot cytoskeleton protein F-actin and cell proteins ZO-1 changes. STAT6 phosphorylation levels of intracellular signaling pathways and further application of the Western blot, to explore the role of IL-13 foot cell injury mechanisms. Results: HSMP presence of IL-13 I receptor mRNA and protein expression. Cytoskeletal proteins F-actin and ZO-1 staining showed: IL-13 showed a dose-and time-dependent podocyte damage. 50ng/ml IL-13 for 24 hours to cause obvious damage podocytes. F-actin thinning disorder, gathered to the cell edge; ZO-1 were significantly weaken and fracture. Further study, IL-13 can induce enough intracellular cytokines STAT6 signaling pathway. STAT6 signaling pathway specific inhibitor leflunomide blocking affect connexin foot cytoskeletal protein structure and cell IL-13. Conclusion: foot cell surface IL-13 receptor, IL-13 by binding to its receptor Connexin podocyte injury of the skeletal structure and between cells. Podocyte injury in IL-13 and STAT6 signaling pathway activation. Part II: Triptolide intervention role of IL-13-induced podocyte injury research purposes: treatment of FSGS drug of choice is still glucocorticoids, but the complete remission rate was less than 30%. . Triptolide therapy for glomerular diseases for more than 30 years of history, has a positive meaning in the treatment of FSGS, low-dose hormone combined triptolide can significantly improve the complete remission rate and shorten the time to complete remission. Previous studies also found that the podocyte injury induced by triptolide puromycin (puromycin aminonucleoside, PA) any protective effect. Study suggest that IL-13-mediated podocyte injury may be one of the mechanisms of the pathogenesis of FSGS. Used in this study HSMPs, with IL-13 induced podocyte injury model, Triptolide protection of podocytes, and to explore its mechanism of action. Method: Triptolide pretreatment of cells with different concentrations (1ng/ml, 3ng/ml, 10ng/ml) for 30 minutes, adding IL-13 (50ng/ml)-induced podocyte injury, immunofluorescence analysis podocyte skeleton related proteins F-actin and cell junction protein ZO-1 changes. STAT6 phosphorylation levels of intracellular signaling pathways and further detected by Western blotting method to explore triptolide factors interfere with IL-13 induced podocyte mechanism. Selection of dexamethasone (1μM) as a drug control. Results: Triptolide significant protective effect of IL-13-induced podocyte injury, similar to its role with dexamethasone. Cytoskeletal proteins F-actin staining results: Triptolide 3ng/ml and 10ng/ml can significantly block the IL-13-induced damage of the F-actin and F-actin expression was partially restored, cytoskeletal rearrangement. ; 1 μM land dexamethasone enable the increased expression of the cytoskeletal protein, cytoskeleton rearrangement. ZO-1 staining results showed that: Triptolide 3ng/ml and 10ng/ml significantly blocked IL-13-induced damage ZO-1, ZO-1 expression increased continuity is restored; the 1 μM ground plug betamethasone can block the IL-13-induced injury, ZO-1 expression and arrangement basically returned to normal ZO-1. Further research also showed that Triptolide and Dexamethasone can inhibit the cytokines IL-13-induced podocyte STAT6 signaling pathway, in which the effect of dexamethasone stronger. Conclusion: Triptolide in vitro significant protective effect of IL-13-induced podocyte injury, this protective effect may be related to the inhibition of IL-13-induced STAT6 signaling pathway activation.

Related Dissertations

  1. Based on the tutor clinical experience in medical treatment of chronic kidney disease proteinuria,R277.5
  2. Study of Triptolide Induced Hepato-toxicology by MS-based Metabonomics Approaches,R285
  3. Clinical and Pathological Characteristics and Risk Factors Associated with Renal Lesions of 91 Patients with Mild Asymptomatic Proteinuria or/and Hematuria,R692
  4. KMSHT active ingredient extraction method optimization and HPLC Fingerprints,R284.1
  5. Prognosis in patients with chronic heart failure multivariate analysis,R541.6
  6. The Th17/treg Regulation of Triptolide on Trinitrobenzene Sulfonic Acid-induced Colitis in Mouse,R285.5
  7. Expression of TrkB in Malignant Melanoma and the Effects of Triptolide on Its Expression,R739.5
  8. Knock-down of Cullin-5 Expression on Alteration of Morphology and Proliferation Ability in Glomerular Podocytes,R726.9
  9. Xinjiang Uygur interleukin- 13 gene polymorphisms and susceptibility to chronic obstructive pulmonary disease association studies,R563.9
  10. Professor Zhang Wenkai Chinese medicine treatment of chronic glomerulonephritis and proteinuria experience,R277.5
  11. QiLingGuBenYiQiTang in Favor of Yiqi Decoction "in the Treatment of Chronic Glomerulonephritis Proteinuria (Spleen and Kidney Yang) Clinical Research,R277.5
  12. The Experimental Research of Thetriptolide and Hydrocortisone Anti-inflammatory Effects,R96
  13. The Effects and Mechanisms of Triptolide on Cell Cycle and Apoptosis in Human Multiple Myeloma Rpmi-8226 Cells,R733.3
  14. Children with nephrotic proteinuria and urinary podocyte nephrin related molecules detection,R726.9
  15. TP on the AIA rat VEGF, FLT1 and CCR5 expression,R684
  16. Triptolide Induces Apoptosis of Hut102 Cells Through P38 MAPK Pathway,R285.5
  17. Granules Compound Gushen senile diabetic nephropathy Kidney Blood type proteinuria clinical observation,R277.5
  18. Triptolide on the experimental study of the role of endometrial cancer cell lines,R737.33
  19. Study in the Anti-Tumor Effect of Triptolide on Human Endometrial Carcinoma in Nude Mice,R737.33
  20. Effect of HGF on the Expression of ILK in Rat Model of Minimal Change Nephrology,R-332
  21. Hepatitis B Virus X Protein Upregulate the Expression of CD59 and Crry in Mouse Podocytes,R692

CLC: > Medicine, health > Chinese Medicine > Of Pharmacy > Pharmacology > Chinese medicine Experimental Pharmacology
© 2012 www.DissertationTopic.Net  Mobile