肝星形细胞CD14表达变化在内毒素肝损伤中的作用

目的 观察内毒素肝损伤过程中肝星形细胞 (枯否细胞 ,KC) CD14表达及肿瘤坏死因子、白细胞介素 - 6的动态变化 ,进而探讨内毒素肝损伤的机理。方法 经尾静脉内注射不同剂量 (1mg/ kg、10 mg/ kg)大肠埃希菌内毒素 (L PS) ,复制内毒素肝损伤模型 ;采用免疫组化方法观察小鼠肝脏 CD14表达变化 ;酶联免疫吸附法 (EL ISA)测定肝组织 TNFα、IL - 6的水平 ,光镜观察肝组织学变化。结果 内毒素肝损伤过程中 ,KC表面 CD14表达呈进行性上调 ,且与内毒素呈明显的量效关系 ;CD14表达的平均光密度值 (OD值 )与肝组织 TNFα、IL - 6及血浆丙氨酸氨基转移酶 (AL T)、总胆红素 (TBIL )呈显著的正相关。结论 内毒素肝损伤过程中 ,随着 CD14表达上调 ,内毒素对库普弗细胞的激活作用则相应增强 ,释放促炎因子 TNFα、IL - 6等进一步增加 ,加重肝组织的损伤 ;肝星形细胞 CD14表达上调可能是内毒素肝损伤的重要机理之一。

创伤弧菌脓毒症大鼠肺组织内毒素受体CD14的基因表达及抗菌药物的干预效应

目的探讨创伤弧菌脓毒症大鼠肺组织内毒素受体CD14的基因表达以及头孢哌酮钠和乳酸左旋氧氟沙星联合抗炎治疗对其的干预效应。方法清洁级雄性SD大鼠随机分为11组。第1组：正常对照组（NC组）；第2～6组：创伤弧菌脓毒症组（VV组），于注射创伤弧菌后不同时点活杀；第7～11组：创伤弧菌脓毒症联合抗炎治疗干预纽（AA组），注射创伤弧菌后6h予腹腔注射头孢哌酮钠和乳酸左旋氧氟沙星，在注射创伤弧菌后不同时点活杀。尤菌取部分肺组织，采用RT—PCR法测定各组CD14mRNA的表达量。比较各组CD14mRNA的表达量。结果VV组各组大鼠肺组织CD14mRNA表达量均明显高于NC组（均P〈0．05）；AA组大鼠染菌后肺组织CD14mRNA表达量呈逐渐降低趋势，与同时点的VV组相比，AA组大鼠肺组织CD14mRNA表达量均明显降低（均P〈0．05）。结论创伤弧菌脓毒症大鼠肺组织CD14mRNA表达量明显增高，及早使用头孢哌酮钠和乳酸左旋氧氟沙星可明显减低创伤弧菌脓毒症大鼠肺组织CD14mRNA的表达水平，有利于抑制炎症反应。

脑缺血模型并发多器官功能障碍综合征时CD14mRNA的表达

目的 探讨脑缺血模型内毒素受体CD1 4mRNA在肺、肝、肠和肾组织的表达变化规律及并发多器官功能障碍综合征 (MODS)的机制。方法 通过阻断双侧颈总动脉 30min建立急性前脑缺血模型 ,随机将 54只大鼠分为正常对照组、假手术组及缺血后 5个亚组 ( 1 2h、2 4h、36h、4 8h、72h组 ) ,依据全身炎症反应综合征 (SIRS)、MODS的诊断标准判断SIRS、MODS的发生率 ,采用原位杂交技术检测缺血后肺、肝、肠和肾脏器CD1 4的基因表达。结果 大鼠急性前脑缺血后SIRS发生率为 1 0 0 % ,MODS发生率为 53 1 % ;缺血后 1 2h、2 4h、36h、4 8h、72h时相点动物肺、肝、肠和肾组织均有不同程度的损害 ,以肺脏和小肠改变为著 ;缺血后 1 2h肺、肝、肠和肾组织CD1 4mRNA表达升高 ,2 4～ 36h达高峰 ,4 8h后下降 ,以肺脏变化最显著 (P <0 0 0 1 ) ;正常对照组和假手术组中肺、肝、肠和肾组织CD1 4mRNA均有不同程度的表达 ,其中两组肺脏CD1 4mRNA的表达有显著性差异 (P <0 0 0 1 )。结论 脑缺血后肺、肠、肝和肾组织CD1 4mRNA的异常表达和病理改变为肠道内毒素易位和内毒素血症的发生提供条件 。

Association of promoter polymorphism of the CD14 C (-159) T endotoxin receptor gene with chronic hepatitis B

AIM: To investigate whether single-nucleotide polymor- phisms in the promoter regions of endotoxin-responsive genes CD14 C (-159) T is associated with chronic hepatitis B. METHODS: We obtained genomic DNA from 80 patients with established diagnosis of chronic hepatitis B and 126 healthy subjects served as a control population. The CD 14 C (-159) T polymorphism was investigated using an allele specific PCR method. RESULTS: Twenty seven percent of chronic hepatitis B patients and 75% of controls were heterozygous for CT genotype. The difference between the chronic hepatitis B and control groups was statistically significant [P < 0.0001; Odds ratio (OR) = 2.887; 95% CI: 1.609-5.178]. Twenty four point six percent of chronic hepatitis B and patients 12.3% of the control group were heterozygous for TT genotype. The difference between groups was not statistically significant (P = 0.256; OR = 0.658; 95% CI: 0.319-1.358). Forty eight point four percent of chronic hepatitis B patients and 12.7% of control were homozy- gote for CC genotype (P < 0.004; OR = 0.416; 95% CI: 0.229-0.755). The frequency of allele C was 61.9% and allele T was 38.1% in hepatitis B patients group. The frequency of allele C was 55.2% and allele T was 44.8% for the control group (P = 0.179; OR = 1.319; 95% CI: 0.881-1.977). CONCLUSION: The TT heterozygous genotype was not a risk factor for chronic hepatitis B. CC homozygote genotype is protective for hepatitis B. Lack of heterozy- gosis of genotype CT is a risk factor for chronic hepatitis B. Alleles C or T were not risk factors for chronic hepatitis B. These findings show the role of a single-nucleotide polymorphism at CD14/-159 on the development ofchronic hepatitis B. Endotoxin susceptibility may play a role in the pathogenesis of chronic hepatitis B.

创伤弧菌脓毒症大鼠肝组织CD14和促／抗炎细胞因子基因表达及抗生素干预研究

目的探讨创伤弧菌脓毒症大鼠肝组织内毒素受体CD14和促／抗炎细胞因子（TNF-α/IL-10）的基因表达及抗菌药物头孢哌酮钠联用乳酸左旋氧氟沙星的干预效应。方法注射创伤弧菌构建创伤弧菌脓毒症模型及药物干预模型，RT—PCR检测大鼠肝组织CD14、TNF-α和IL-10的基因表达水平。结果与正常对照组（NC组）相比，创伤弧菌脓毒症组（VV组）感染细菌2、6、9、12、16h的CD14 mRNA和TNF-α mRNA表达量均明显升高（P〈0．05），IL-10 mRNA表达量在感染后9、12、16h也明显升高（P〈0．05）。创伤弧菌脓毒症联用抗生素干预组（AA组）CD14 mRNA表达在感染后9h，TNF-αmRNA表达在感染后9、12h，IL-10 mRNA表达在感染后9、12、16h仍明显高于NC组（P〈0．05）。与相同时间点的VV组相比，AA组感染后9、12、16h的CD14 mRNA表达量，16h的TNF-α mRNA和IL-10 mRNA表达量均明显降低（P〈0．05）。结论创伤弧菌脓毒症大鼠肝组织CD14 mRNA表达在脓毒症早期即达高峰，脓毒症过程中表现为持续增高，脓毒症早期肝组织TNF-α mRNA明显升高，脓毒症中后期肝组织IL-10 mRNA才逐渐增多。及早使用头孢哌酮钠和乳酸左氧氟沙星可明显减低创伤弧菌脓毒症大鼠肝组织CD14 mRNA、TNF-α mRNA和IL-10 mRNA表达水平。头孢哌酮钠联用乳酸左氧氟沙星可显著减少创伤弧菌脓毒症肝组织的内毒素受体的表达，有利于机体致炎／抗炎平衡恢复。

Innate immune reactivity of the liver in rats fed a choline-deficient L-amino-acid-defined diet

AIM： To investigate the innate immune reactivity of tumor necrosis factor-alpha （TNF-α）, Toll-like receptor 4 （TLR4）, and CD14 in the liver of non-alcoholic steatohepatitis （NASH） model rats. METHODS： Male F344 rats were fed a cholinedeficient L-amino-acid-defined （CDAA） diet. The rats were killed after 4 or 8 wk of the diet, and their livers were removed for immunohistochemical investigation and RNA extraction. The liver specimens were immunostained for TNF-α, TLR4, and CD14. The gene expressions of TNF-α, TLR4, and CD14 were determined by reverse-transcriptase polymerase chain reaction （RT-PCR）. Kupffer cells were isolated from the liver by Percoll gradient centrifugation, and were then cultured to measure TNF-α production. RESULTS： The serum and liver levels of TNF-~ in the CDAA-fed rats increased significantly as compared with the control group, as did the immunohistochemical values and gene expressions of TNF-α, TLR4, and CD14 with the progression of steatohepatitis. TNF-α production from the isolated Kupffer cells of the CDAAfed rats was elevated by lipopolysaccharide stimulation. CONCLUSION： The expressions of TNF-α, TLR4, and CD14 increased in the NASH model, suggesting that