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Peripheral Blood Mononuclear Cell Expression of Toll-like Receptors and Relation to Endotoxin Levels in HBV-associated Cirrhosis

Author: PanQinCong
Tutor: ZhangShunCai
School: Fudan University
Course: Internal Medicine
Keywords: Endotoxin Endotoxemia Toll-like receptor Mononuclear cells Cirrhosis Tumor Necrosis Factor
CLC: R575.2
Type: Master's thesis
Year: 2008
Downloads: 121
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Abstract


Background and Purpose: LPS (endotoxin) present in the cell wall of Gram-negative bacteria, lipopolysaccharide complexes (lipopolysaccharides, LPS), a component of the main pathogenic Gram-negative bacteria, bacterial endotoxins can cause internal endotoxemia. Patients with cirrhosis complicated by endotoxemia, and endotoxin levels in the blood and the degree of liver damage. Bacterial infection or endotoxin-induced inflammation in patients with liver cirrhosis severity of natural immunity, the body's natural immunity against external pathogens first line of defense. Recent data show that mammalian Toll-like receptors (toll-like receptors, TLR) participation by endotoxin or other pathogen-induced intracellular signal transduction, the most critical is the TLR2 and TLR4 molecules. In congenital immunity, TLR is seen as anti-microbial reactions triggered a key ingredient. Therefore, this study aims to: 1) detection of hepatitis B patients with cirrhosis peripheral blood mononuclear cells (Peripheral blood mononuclear cell, PMBC) TLR2 and TLR4 mRNA expression; 2) to explore TLR2/TLR4 mRNA expression and serum endotoxin and tumor necrosis factor-α (tumor necrosisfactor-α, TNF-α) in relation to clarify TLR2/TLR4 endotoxemia in liver cirrhosis in rats. Methods: 29 patients enrolled patients with liver cirrhosis in 2001 are in line with the diagnostic criteria for viral hepatitis and cirrhosis. Except for the following: (a) nearly six weeks used immunomodulatory drugs; (2) renal insufficiency (creatinine> 120 umol / L): (3) combined with tumor patients; (4) no history of drinking (daily alcohol less than 20g); (5) 6 weeks, no clear infection or antibiotics. The extent of damage to liver function was graded Child-Pugh criteria. Select by sex and age-matched healthy volunteers without a history of hepatitis 10 cases of normal control, requiring liver function tests normal, no history of drinking. Patients with peripheral blood extract 7 ml, 5 ml indwelling wherein the enzyme treatment in the de anticoagulant tube, using lymphocyte separation medium (Ficoll) isolated mononuclear cells; using semi-quantitative RT-PCR of TLR2 and TLR4 mRNA levels; another 2ml indwelling in the absence of endotoxin in vitro separation of serum, separated serum stored at -70 ℃ in the refrigerator, with the quantitative determination of serum LAL endotoxin levels and the use of radioimmunoassay of serum TNF-α levels. Results: (1) the level of serum endotoxin and serum TNF-α level changes: compared with normal controls, patients with cirrhosis serum endotoxin levels (respectively 0.050EU/ml and 0.107EU/ml P <0.05) and serum TNF- α levels (respectively: 1.282pg/ml and 2.076pg/ml, P <0.05) were significantly higher; (2) in peripheral blood mononuclear cells receptors TLR2 and TLR4 mRNA expression: compared with normal controls, cirrhosis peripheral blood mononuclear cells was significantly increased TLR2 (respectively: 0.651 and 0.923, P <0.05), and with serum endotoxin and TNF-α was positively correlated with elevated levels (r = 0.573), while the expression of TLR4 down, but the difference was not statistically significant. Conclusions: In patients with liver cirrhosis (Child-Pugh B is mainly level, Child-Pugh C grade) PMBCs of TLR2 mRNA expression levels increased, and with TNF-α levels were positively correlated, but no change in the level of TLR4 mRNA, indicating endotoxin present in patients with cirrhosis resistance phenomenon, there may be both Gram-positive bacteria by TLR2-mediated displacement cytokine release.

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CLC: > Medicine, health > Internal Medicine > Digestive and abdominal diseases > Liver and gall bladder disease > Cirrhosis
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