Dissertation > Excellent graduate degree dissertation topics show

Expression of TIPE2, TLR2, TLR4mRNA in PBMCs of Patients with Chronic Hepatitis C Virus

Author: LiuKun
Tutor: KongLi
School: Hebei Medical University
Course: Traditional Chinese Medicine
Keywords: TNFAIP8L2(TIPE2) TLR2 TLR4 T-lymphocyte Subsets chronic hepatitis C(CHC) peripheral blood mononuclear cells (PBMCs) immune homeostasis
CLC: R512.63
Type: Master's thesis
Year: 2012
Downloads: 75
Quote: 0
Read: Download Dissertation

Abstract


Objective:Hepatitis C virus (HCV), a single-stranded positive-sense RNA virus, has a length of~9.6kb. And it is estimated that3%of the world’s population are infected with HCV, The majority of those (50-85%) may leads to chronic liver inflammation, eventually progress to liver cirrhosis or hepatocellular carcinoma. Until recently, it has been believed that host immune responses at the cellular level and adaptive immune responses are prerequisite for limiting HCV infection and causing cell death in the pathological process. But, the immune mechanism understanding the pathogenesis of hepatic inflammation induced by HCV is still enigmatic. TIPE2, the tumor necrosis factor-α-induced protein8-like2(TNFAIP8L2), is a newly identified negative regulator of innate and adaptive immunity. Similarly to other TNFAIP8family, TIPE2plays a crucial role in maintaining immune homeostasis. Initial studies demonstrated that by negatively regulating T cell receptor (TCR) and Toll-like receptor (TLR), TIPE2governing both the innate and adaptive immune systems. As a negative inhibitor, TIPE2was detected in peripheral blood and tissue from patients with many diseases, such as chronic hepatitis B, systemic lupus erythematosus, rheumatoid arthritis, colon cancer, etc. Toll-like receptors, nucleotide-binding oligo-dimerisation domain (NOD)-like receptors, recognize pathogenassoci-ated molecular patterns found in bacteria, viruses, fungi and protozoa and damage-associated molecular patterns to facilitate innate and adaptive immunity for maintaining immune homeostasis. In this study, we examined the mRNA expression levels of TIPE2, TLR2and TLR4in peripheral blood mononuclear cells (PBMCs) from chronic hepatitis C patients, and aimed to elucidate whether these genes participate in the pathogenesis of HCV-induced hepatitis and the interaction between TIPE2and TLRs in hepatic inflamma- tionMethods:1Human subjectsThis study consisted of60chronic hepatitis C (CHC) patients (Male/female,22/38) with an average age of43.51±14.90years, who were admitted to Department of Traditional and Western Medical Hepatology of The Third Hospital of He Bei Medical University, from December2009to October2011. There was no serologic evidence of co-infection with other hepatotropic viruses. The study was conducted according to the criteria from the Prevention and Therapy Regimen of Viral Hepatitis revised at Xian Conference in2000and the Prevention and Therapy Regimen of Viral Hepatitis revised diagnostic criteria in2004.30healthy controls (Male/female,13/17) were also recruited with an average of39.23±11.53years.7ml of anticoagulation blood from all procedures was used to separate peripheral blood mononuclear cells, and5ml of vein blood was obtained to separate serum.2Treatment methodsInterferon plus ribavirin were used to antiviral therapy. According to decreased degree of leukocyte and Neutrophil, common drugs of promoting leukocyte were administered.3Detection of the mRNA expression levels of TIPE2, TLR2and TLR4in PBMCs from CHC patientsBlood was added to acid citrate dextrose anticoagulant and separated by Ficoll density gradient centrifugation and then stored in Trnzol reagent to stabilize the RNA. Total RNA was subjected to RNase free DNase to remove genomic DNA contamination. We used ultraviolet spectrophotometer to detect of the purity and concentration of RNA, and elected the absorbance (A) on260/280nm is between1.8-2.0. cDNA was synthesized using5units of Reverse Transcription System. Primer Premier5.0software was used to design specific primers for the TIPE2fragments. The mRNA expression levels of TIPE2, TLR2and TLR4were evaluated by RT-PCR.4Serum levels of clinical parameters of CHC We collected serums from CHC patients and used ELISA to test anti-HCV and automatic biochemistry analyzer to determine serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (Tbil) levels. Detection of HCV RNA was performed with qualitative RT-PCR.5Detection of peripheral blood T lymphocyte subsetsWe collected2ml fresh anticoagulant blood from CHC patients, and used flow cytometry to determine the levels of CD3+, CD4+and CD8+T lympho-cyte.Results:1The mRNA expression levels of TIPE2in PBMCs from CHC patients and the correlation with antiviral therapyWe examined the mRNA expression levels of TIPE2in peripheral blood mononuclear cells of the HCV infected patients and healthy controls. The TIPE2mRNA expression was dramatically reduced in HCV patients compared with healthy controls (0.49±0.17vs1.06±0.30, P<0.05). In42patients with interferon plus ribavirin antiviral therapy, the TIPE2mRNA expression in PBMCs was evidently increased than pre-treatments (0.41±0.11vs.0.66±0.25,P<0.01).2The mRNA expression levels of TLR2in PBMCs from CHC patients and the correlation with antiviral therapyThe mRNA levels of TLR2was significantly higher in peripheral blood mononuclear cells from chronic hepatitis C patients than in healthy controls (1.86±0.35vs1.24±0.30, P<0.05);42patients after interferon plus ribavirin antiviral therapy, the TLR2mRNA in PBMCs evidently decreased than pre-treatments (TLR2,1.94±0.33vs1.37±0.19, P<0.01).3The mRNA expression levels of TLR4in PBMCs from CHC patients and connection with antiviral therapyRT-PCR analysis showed that the mRNA levels of TLR4was significantly higher in peripheral blood mononuclear cells from chronic hepatitis C patients than in healthy controls (1.86±0.35vs1.24±0.30, P<0.05);42patients after interferon plus ribavirin antiviral therapy, the TLR4mRNA in PBMCs evidently decreased than pre-treatments (1.58±0.56vs1.16±0.15, P<0.01).4Correlation between mRNA expression levels of TIPE2and clinical parame-ters of CHCWe analyzed clinical parameters of these HCV patients, according to the serum levels of HCV RNA, patients received no immunomodulatory treatment were subdivided into HCV RNA low group (HCV RNA≤1×104copies/mL) and HCV RNA high group (HCV RNA>1×104copies/mL). Similarly, based on the serum levels of ALT, AST and Tbil, HCV patients were also subdivided into normal and high level groups. Statistical analysis results showed that the mRNA expression levels of TIPE2in peripheral blood mononuclear cells in the low level groups was much higher than that in the high level groups (HCV RNA,0.66±0.16vs0.44±0.11, P<0.05; ALT,0.64±0.13vs0.38±0.09, P<0.05; AST,0.64±0.15vs0.37±0.09, P<0.05; Tbil,0.58±0.14vs0.34±0.07, P<0.01), and the mRNA expression levels of TIPE2was negatively correlated with clinical parameters of HCV (HCV RNA, r=-0.658, P<0.01; ALT, r=-0.912, P<0.01; AST, r=-0.839, P<0.01), however, there was no direct correlation between TIPE2mRNA levels with Tbil activities.5Correlation between mRNA expression levels of TIPE2, TLR2and TLR4Furthermore, SPSS13.0analysis showed there was a negative correlation between the mRNA expression levels of TIPE2and TLR2, TLR4(TLR2, r=-0.669, P<0.01; TLR4, r=-0.649, P<0.01).6Variation of T lymphocyte subsets from CHC patients’peripheral blood and the relationship between TIPE2and T subsetsIt was demonstrated that significantly lower percentage CD3+and CD4+T lymphocyte, and higher of CD8+T lymphocyte, with lower CD4+/CD8+T lymphocyte ratio in HCV patients than that in healthy controls(CD3+,64.28±10.39vs70.97±3.80, P<0.05;CD4+,33.60±5.26vs44.66±3.62, P<0.05; CD8+,29.59±6.47vs25.63±4.03, P<0.05; CD4+/CD8+,1.19±0.35vs1.78 ±0.34, P<0.05). There was a positive correlation between the mRNA expression levels of TIPE2and CD4+T lymphocyte and CD4+/CD8+(CD4+r=0.735, P<0.01; CD4+/CD8+, r=0.804, P<0.01), and negative correlation between the mRNA expression levels of TIPE2and CD8+T lymphocyte (r=-0.625, P<0.01), there was no correlation between the levels of TIPE2and CD3+Conclusion:1Patients with chronic hepatitis C virus had evidently reduced levels of TIPE2expression in peripheral blood mononuclear cells compared to healthy individuals, and after antiviral and anti-flammation therapy this levels was significantly increased.2The mRNA levels of TLR2, TLR4was much higher on Peripheral blood mononuclear cell in patients with chronic hepatitis C than those in healthy individuals, and after antiviral and anti-flammation therapy these levels was significantly reduced.3Down-regulation of TIPE2mRNA expression may result in elevated hepatic inflammation and fibrosis, and TIPE2was negatively correlated with HCV RNA.4TIPE2may involve in the pathogenesis of HCV-induced hepatitis and regulate the cellular immunity, determine the progression of chronic hepatitis C.

Related Dissertations

  1. The Polymorphisms of TLR4 Gene and Function Analysis of C1027A in Suzhong Pig,S828
  2. The Relationship between TLR4 Protein Expression of Peripheral Blood Monocytes and the Pathogenetic Condition and Prognosis in Patients with Acute Coronary Syndrome,R541.4
  3. The Effect of Mild Hypothermia Combined with TP-5 to T Lymphocyte Subsets and Prognosis of Patients with Severe Traumatic Brain Injur,R651.15
  4. Study for Triptolide in Protection Against Lps-induced Acute Lung Injury in Rats,R285.5
  5. The Role of Toll-like Receptors 2 and 4 in Cardiac Allograft Rejection,R654.2
  6. Experimental Study the Neuroprotective Role of Progesterone on Early Brain Injury in Male Rats after Subarachnoid Hemorrhage,R743.35
  7. The Expression of TLR2 and Effect on Invasion Ability in Gastric Cancer,R735.2
  8. Probiotics on Experimental Colitis Toll-like Receptor 4 Expression in Rats,R574.62
  9. The Protective Effects of Glutamine in Human Bronchial Epithelial Cells Stimulated by Lipopolysaccharide Administration,R96
  10. Effects of TLR2 on IL-8 and γ-IFN Secretion of Keratinocytes in Dermatophytic Infections,R756
  11. LPS, tumor necrosis release substances on B16 mouse melanoma cell invasion and metastasis ability of,R739.5
  12. Establishment of Immature Rat MTLE Model and the Study of TLR4, IL-1β Expression Changes in MTLE,R742.1
  13. The Expression of Toll-like Receptor 4 in the Endometrium of Patients with IUAs,R363
  14. Alleviative Effect of Arginine on Liver Injury of Piglets After Lipopolysaccharide Challenge and Its Mechanism,S828.5
  15. Experimental Study of Cell Immune and Intestinal Barrier After Severe Trauma - Hemorrhagic Shock in Rats,R641
  16. Integrative Medicine Immunity During Early Pregnancy Recurrent Abortion Retrospective Analysis,R714.21
  17. The Effect of PPA on the Expression of TLR4、Act1、A20 and Proliferation in Human Bronchial Epithelial Cells,R965
  18. The Immune Inflammatory Response Early after Cerebral Ischemia Reperfusion in Mice and the Intervention of Albumin or Zinc Chloride,R743.3
  19. Preliminary Exploration of the Effect of Caveolin-1 in the TLR4-mediated Inflammatory Signaling Pathways Stimulated by LPS in Human Breast Epithelial Cell,R655.8
  20. Expression and Regulation of a Novel Identified TNFAIP8 Family is Associted with Diabetic Nephropathy,R587.2
  21. The Effects and Mechanisms of Microgravity on Expression of TF in Monocytes,R85

CLC: > Medicine, health > Internal Medicine > Infectious disease > Viral infections > Viral Hepatitis > Hepatitis C (non- A non-B )
© 2012 www.DissertationTopic.Net  Mobile