中华传染病杂志
中華傳染病雜誌
중화전염병잡지
CHINESE JOURNAL OF INFECTIOUS DISEASES
2009年
3期
133-137
,共5页
庄焱%谢青%晏春根%王晖%蔡伟%林兰意%安宝燕%刘芸野%周霞秋%俞红%郭清
莊焱%謝青%晏春根%王暉%蔡偉%林蘭意%安寶燕%劉蕓野%週霞鞦%俞紅%郭清
장염%사청%안춘근%왕휘%채위%림란의%안보연%류예야%주하추%유홍%곽청
受体,细胞表面%肝硬化%肿瘤坏死因子α
受體,細胞錶麵%肝硬化%腫瘤壞死因子α
수체,세포표면%간경화%종류배사인자α
Receptors,cell surface%Liver cirrhosis%Tumor necrosis factor-alpha
目的 探讨乙型肝炎(乙肝)肝硬化患者中Toll样受体(TLR)2、4表达与细胞因子的相关性.方法 采用随机数字表法随机收集35例乙肝肝硬化患者与35例健康对照者的静脉抗凝血.ELISA检测血TNF-α含量.分离两组外周血单个核细胞(PBMC),流式细胞仪检测PBMC表面TLR2、4表达.实时荧光定量PCR检测每份PBMC中TLR2、4 mRNA的表达水平.正态分布均数比较采用t检验和单因素方差分析,非正态分布数据采用Mann-Whitney U检验、Kruskal-Wallis H检验和Spearman相关分析.结果 肝硬化组的血TNF-α水平明显高于对照组(33.52 ng/L比6.07 ng/L,Z=6.584,P<0.01),且随着Child-Pugh评分的增加而升高.肝硬化组PBMC中TLR2阳性细胞率明显高于对照组(20.65%比12.04%,Z=-4.458,P<0.01),与血TNF-α水平存在正相关(r=0.448 3,P<0.05),且随着Child-Pugh评分的增加而升高.两组PBMC中TLR4阳性率差异无统计学意义.肝硬化组PBMC中TLR2/还原型磷酸甘油醛脱氢酶(GAPDH)mRNA表达水平明显高于对照组(0.234 2比0.043 1,Z=-6.83,P<0.01),与血TNF-α水平存在正相关(r=0.411 1,P<0.05),且随着Child-Pugh评分的增加而升高.两组PBMC中TLR4 mRNA表达水平差异无统计学意义.结论 肝硬化患者PBMC中TLR2表达水平明显增高,与血TNF-α水平及肝硬化的严重程度呈正比,但TLR4表达水平无明显变化,提示在乙肝肝硬化患者中,是TLR2而非TLR4在肝硬化的进展过程中具有重要作用.
目的 探討乙型肝炎(乙肝)肝硬化患者中Toll樣受體(TLR)2、4錶達與細胞因子的相關性.方法 採用隨機數字錶法隨機收集35例乙肝肝硬化患者與35例健康對照者的靜脈抗凝血.ELISA檢測血TNF-α含量.分離兩組外週血單箇覈細胞(PBMC),流式細胞儀檢測PBMC錶麵TLR2、4錶達.實時熒光定量PCR檢測每份PBMC中TLR2、4 mRNA的錶達水平.正態分佈均數比較採用t檢驗和單因素方差分析,非正態分佈數據採用Mann-Whitney U檢驗、Kruskal-Wallis H檢驗和Spearman相關分析.結果 肝硬化組的血TNF-α水平明顯高于對照組(33.52 ng/L比6.07 ng/L,Z=6.584,P<0.01),且隨著Child-Pugh評分的增加而升高.肝硬化組PBMC中TLR2暘性細胞率明顯高于對照組(20.65%比12.04%,Z=-4.458,P<0.01),與血TNF-α水平存在正相關(r=0.448 3,P<0.05),且隨著Child-Pugh評分的增加而升高.兩組PBMC中TLR4暘性率差異無統計學意義.肝硬化組PBMC中TLR2/還原型燐痠甘油醛脫氫酶(GAPDH)mRNA錶達水平明顯高于對照組(0.234 2比0.043 1,Z=-6.83,P<0.01),與血TNF-α水平存在正相關(r=0.411 1,P<0.05),且隨著Child-Pugh評分的增加而升高.兩組PBMC中TLR4 mRNA錶達水平差異無統計學意義.結論 肝硬化患者PBMC中TLR2錶達水平明顯增高,與血TNF-α水平及肝硬化的嚴重程度呈正比,但TLR4錶達水平無明顯變化,提示在乙肝肝硬化患者中,是TLR2而非TLR4在肝硬化的進展過程中具有重要作用.
목적 탐토을형간염(을간)간경화환자중Toll양수체(TLR)2、4표체여세포인자적상관성.방법 채용수궤수자표법수궤수집35례을간간경화환자여35례건강대조자적정맥항응혈.ELISA검측혈TNF-α함량.분리량조외주혈단개핵세포(PBMC),류식세포의검측PBMC표면TLR2、4표체.실시형광정량PCR검측매빈PBMC중TLR2、4 mRNA적표체수평.정태분포균수비교채용t검험화단인소방차분석,비정태분포수거채용Mann-Whitney U검험、Kruskal-Wallis H검험화Spearman상관분석.결과 간경화조적혈TNF-α수평명현고우대조조(33.52 ng/L비6.07 ng/L,Z=6.584,P<0.01),차수착Child-Pugh평분적증가이승고.간경화조PBMC중TLR2양성세포솔명현고우대조조(20.65%비12.04%,Z=-4.458,P<0.01),여혈TNF-α수평존재정상관(r=0.448 3,P<0.05),차수착Child-Pugh평분적증가이승고.량조PBMC중TLR4양성솔차이무통계학의의.간경화조PBMC중TLR2/환원형린산감유철탈경매(GAPDH)mRNA표체수평명현고우대조조(0.234 2비0.043 1,Z=-6.83,P<0.01),여혈TNF-α수평존재정상관(r=0.411 1,P<0.05),차수착Child-Pugh평분적증가이승고.량조PBMC중TLR4 mRNA표체수평차이무통계학의의.결론 간경화환자PBMC중TLR2표체수평명현증고,여혈TNF-α수평급간경화적엄중정도정정비,단TLR4표체수평무명현변화,제시재을간간경화환자중,시TLR2이비TLR4재간경화적진전과정중구유중요작용.
Objective To explore the relationship among cytokine levels and Toll-like receptor (TLR) 2,4 in hepatitis B virus (HBV) related cirrhosis. Methods Heparin anticoagulated venous blood of 35 randomly selected HBV related cirrhosis and 35 healthy volunteers were collected aseptically. Plasma tumor necrosis factor (TNF)-α concentration was determined using enzyme-linked immunosorbent assay (ELISA). Peripheral blood mononuclear cells (PBMC) were separated and stained with anti-TLR2,4 monoclonal antibodies, then analysed by flow cytometry. Total RNA was extracted from PBMC and TLR2,4 mRNA expression levels were evaluated by real-time quantitative polymerase chain reaction (PCR) using SYBR Green detection. Means of normal distribution were compared by t test and one factor analysis of variance. The data of abnormal distribution were analyzed using Mann-Whithey U test, Kruskal-Wallis H test and Spearman correlation analysis. Results The plasma concentration of TNF-α in the cirrhotic patients was significantly higher than that in the healthy controls (33.52 ng/L vs 6.07 ng/L, Z=-6.584, P<0. 01), which was parellel with Child-Pugh score. TLR2 positive rate in PBMC from the cirrhotic patients was significantly higher than that from the healthy controls (20.65% vs 12.04%, Z=-4.458, P<0.01), which was positively correlated with plasma TNF-α level (r= 0.448 3, P<0.05), and parellel with Child-Pugh score. Between the cirrhotic and healthy groups, there was no significant difference of TLR4 positive rate in PBMC. The mRNA expression level of TLR2/GAPDH in PBMC from the cirrhotic patients was significantly higher than the controls (0.234 2 vs 0.043 1, Z=-6.83, P<0.01), which was positively correlated with plasma TNF-α level (r=0.411 1, P<0.05), and parellel with Child-Pugh score. Between the two groups, there was no significant difference of TLR4 mRNA expression level in PBMC. Conclusions The expression of TLR2 in PBMC from cirrhotic patients is significantly elevated, which is positively correlated with plasma TNF-α level and the severity of liver disease. The expression of TLR4 in PBMC from cirrhotic patients is unchanged. It suggests that TLR2 but not TLR4, plays an important role in the progression of liver cirrhosis.
