Analysis of TLR4 and TLR2 polymorphisms in inflammatory bowel disease in a Guangxi Zhuang population

AIM: To study the polymorphisms of toll-like receptor 4 (TLR4) gene Asp299Gly, Thr399Ile and TLR2 gene Arg753Gln, Arg677Trp and susceptibility to inflammatory bowel disease (IBD) in the Zhuang population from Guangxi, China.METHODS: A case-control study was performed from February 2007 to October 2011 which included 146 Zhuang patients with IBD in the experimental group and 164 healthy Zhuang subjects who acted as the control group. All patients and healthy subjects were from the Guangxi Zhuang Autonomous Region of China. Genomic DNA was extracted from intestinal tissue by the phenol chloroform method. TLR4 gene Asp299Gly, Thr399Ile and TLR2 gene Arg753Gln, Arg677Trp were amplified by polymerase chain reaction (PCR), and then detected by PCR-restriction fragment length polymorphism (RFLP).RESULTS: The TLR4 gene Asp299Gly was digested using Nco I restriction enzyme, and a single band of 249 bp was observed which showed that it was a wild type (AA). The TLR4 gene Thr399Ile was digested using Hinf Irestriction enzyme and only the wild type (CC) was detected. In addition, the TLR2 gene Arg677Trp was digested using Aci I restriction enzyme and only the wild type (CC) was detected. The TLR2 gene Arg753Gln was digested using Pst I restriction enzyme. Only the wild type (GG) as a single band of 254 bp was observed during RFLP. Overall, no heterozygous or homozygous single nucleotide polymorphism mutations were found in patients with Crohn’s disease and ulcerative colitis both in the TLR4 gene Asp299Gly, Thr399Ile and the TLR2 gene Arg677Trp, Arg753Gln in the Zhuang population from the Guangxi Zhuang Autonomous Region of China.CONCLUSION: The TLR4 gene Asp299Gly, Thr399Ile and TLR2 gene Arg753Gln, Arg677Trp polymorphisms may not be associated with IBD in the Zhuang population from the Guangxi Zhuang Autonomous Region of China.     

mRNA levels of TLR4 and TLR5 are independent of H pylori

AIM：To determine if the presence H pylori or its virulence affect toll-like receptor 4 （TLR4） and TLR5 mRNA expression levels. METHODS：For the in vivo assays, gastric biopsies were obtained from 40 patients and H pylori status was determined. For the in vitro assays, human gastric adenocarcinoma mucosal cells （AGS） were cultured in the presence or absence of twelve selected H pylori strains. H pylori strains isolated from culture-positive patients and selected strains were genotyped for cagA and vacA. The cDNA was obtained from mRNA extracted from biopsies and from infected AGS cells. TLR4 and TLR5 mRNA levels were examined by real-time PCR. RESULTS： The presence of H pylori did not affect the mRNA levels of TLR4 or TLR5 in gastric biopsies. The mRNA levels of both receptors were not influenced by the vacA status （P 〉 0.05 for both receptors） andthere were no differences in TLR4 or TLR5 mRNA levels among the different clinical presentations/histological fi ndings （P 〉 0.05）. In the in vitro assay, the mRNA levels of TLR4 or TLR5 in AGS cells were not influenced by the vacAs1 status or the clinical condition associated with the strains （P 〉 0.05 for both TLR4 and TLR5）. CONCLUSION： The results of this study show that the mRNA levels of TLR4 and TLR5 in gastric cells, both in vivo and in vitro, are independent of H pylori colonization and suggest that vacA may not be a significant player in the first step of innate immune recognition mediated by TLR4 or TLR5.     

TLR4 polymorphisms, infectious diseases, and evolutionary pressure during migration of modern humans

Infectious diseases exert a constant evolutionary pressure on the genetic makeup of our innate immune system. Polymorphisms in Toll-like receptor 4 (TLR4) have been related to susceptibility to Gram-negative infections and septic shock. Here we show that two polymorphisms of TLR4, Asp299Gly and Thr399Ile, have unique distributions in populations from Africa, Asia, and Europe. Genetic and functional studies are compatible with a model in which the nonsynonymous polymorphism Asp299Gly has evolved as a protective allele against malaria, explaining its high prevalence in subSaharan Africa. However, the same allele could have been disadvantageous after migration of modern humans into Eurasia, putatively because of increased susceptibility to severe bacterial infections. In contrast, the Asp299Gly allele, when present in cosegregation with Thr399Ile to form the Asp299Gly/Thr399Ile haplotype, shows selective neutrality. Polymorphisms in TLR4 exemplify how the interaction between our innate immune system and the infectious pressures in particular environments may have shaped the genetic variations and function of our immune system during the out-of-Africa migration of modern humans.     

Adipocytes-derived exosomal miR-122 promotes non-alcoholic fat liver disease progression via targeting Sirt1

AimsNonalcoholic fatty liver disease (NAFLD) is a chronic liver disease that affects adipose function. This study aimed to explore the function of adipocytes-derived exosomal (ADEs) miR-122 in NAFLD.MethodsA high-fat and high-fructose diet-induced rat model and a palmitic acid (PA)-induced in vitro model were established. The RNA level of miR-122 and Sirt1 was measured using qRT-PCR. The protein levels of exosome biomarkers, and lipogenesis, inflammation and fibrosis biomarkers were determined by western blotting. Cell viability and apoptosis were assessed using cell counting kit-8 and flow cytometry, respectively. Serum alanine aminotransferase, aspartate aminotransferase, total cholesterol, triglyceride levels were measured. Liver tissue damage was assessed using haematoxylin and eosin staining. The interaction between miR-122 and Sirt1 3′UTR was assessed using a luciferase reporter gene assay.ResultsADEs exhibited abundant level of miR-122 and promoted lipogenesis, impaired hepatocyte survival, enhanced liver damage and increased serum lipid levels in vivo and in vitro. Inhibition of miR-122 in ADEs alleviated NAFLD progression, lipid and glucose metabolism, liver inflammation and fibrosis both in vivo and in vitro. miR-122 binds directly to the 3′UTR of Sirt1 to suppress its expression. Moreover, Sirt1 overexpression reversed the increase in cell apoptosis, glucose and lipid metabolism, liver inflammation and fibrosis induced by ADEs in vivo and in vitro.ConclusionsThe ADEs miR-122 promotes the progression of NAFLD via modulating Sirt1 signalling in vivo and in vitro. The ADEs miR-122 may be a promising diagnostic biomarker and therapeutic target for NAFLD.     

Overexpression of HMGB1 A-box reduced lipopolysaccharide-induced intestinal inflammation via HMGB1/TLR4 signaling in vitro

AIM: To investigate the inhibitory effects and mechanism of high mobility group box(HMGB)1 A-box in lipopolysaccharide(LPS)-induced intestinal inflammation.METHODS: Overexpression of HMGB1 A-box in human intestinal epithelial cell lines(SW480 cells) was achieved using the plasmid p EGFP-N1. HMGB1 A-box-overexpressing SW480 cells were stimulated with LPS and co-culturing with human monocyte-like cell lines(THP-1 cells) using a Transwell system, compared with another HMGB1 inhibitor ethyl pyruvate(EP). The m RNA and protein levels of HMGB1/toll-like receptor(TLR) 4 signaling pathways [including HMGB1, TLR4, myeloid differentiation factor88(MYD88), Phosphorylated Nuclear Factor κB(p NF-κB) p65] in the stimulated cells were determined by realtime polymerase chain reaction and Western blotting. The levels of the proinflammatory mediators [including HMGB1, interleukin(IL)-1β, IL-6 and tumor necrosis factor(TNF)-α] in the supernatants of the stimulated cells were determined by ELISA.RESULTS: EP downregulated the m RNA and protein levels of HMGB1, inhibited the TLR4 signaling pathways(TLR4, MYD88 and p NF-κB p65) and reduced the secretion of proinflammatory mediators(HMGB1, IL-1β, IL-6 and TNF-α) in the SW480 and THP-1 cells activated by LPS but not in the unstimulated cells. Activated by LPS, the overexpression of HMGB1 A-box in the SW480 cells also inhibited the HMGB1/TLR4 signaling pathways and reduced the secretion of these proinflammatory mediators in the THP-1 cells but not in the transfected and unstimulated cells. CONCLUSION: HMGB1 A-box, not only EP, can reduce LPS-induced intestinal inflammation through inhibition of the HMGB1/TLR4 signaling pathways.     

Polymorphisms of the TLR2 and TLR4 genes are associated with risk of gastric cancer in a Brazilian population

AIM: To investigate toll-like receptor 2 (TLR2) -196 to -174 del, and TLR4 (+896A/G rs4986790 and +1196C/T rs4986791) polymorphisms at risk of chronic gastritis and gastric cancer in a Brazilian population and association of gastric lesions with risk factors such as smoking, alcohol intake and Helicobacter pylori infection.METHODS: In this case-control study, polymorphism at TLR2 -196 to -174 del was investigated by using the allele-specific polymerase chain reaction (PCR) method, while the PCR-restriction fragment length polymorphism technique was carried out to identify the TLR4 (rs4986790 and rs4986791) genotypes in 607 Brazilian individuals (208 with chronic gastritis-CG, 174 with gastric cancer-GC and 225 controls -C).RESULTS: The single nucleotide polymorphisms TLR4+1196C/T was not associated with risk of chronic gastritis or gastric cancer and the homozygous genotypes TLR4+896GG and TLR4+1196TT were absent in the studied population. However, the frequency of TLR2 -196 to -174 ins/del + del/del and TLR4+896AG genotypes was significantly higher (P < 0.01 and P = 0.01, respectively) in the cancer group (33.4% and 11.5%, respectively) than in the control group (16.9% and 4.5%, respectively). It was also observed that the G-C haplotype of the TLR4+896A/G+1196C/T (P = 0.02) and the combination of variant alleles of the TLR2/TLR4+896G (P = 0.02) are associated with susceptibility to gastric cancer. In addition, the multiple logistic regression showed that male gender [odds ratio (OR) = 2.70; 95% CI: 1.66-4.41; P < 0.01], alcohol intake (OR = 2.93; 95% CI: 1.76-4.87; P < 0.01), TLR2 -196 to -174 del (OR = 2.64; 95% CI: 1.56-4.44; P < 0.01) and TLR4+896G (OR = 3.19; 95% CI: 1.34- 7.61; P < 0.01) polymorphisms were associated with a higher susceptibility to developing this neoplasm.CONCLUSION: Our data indicate that TLR2 -196 to -174 del and TLR4+896G may increase the risk of gastric cancer in a Brazilian population.     

TLR4基因Asp299Gly及TLR2基因Arg753Glu、Arg677Trp多态性与中国湖北汉族炎症性肠病无相关性

目的:研究TLR4基因Asp299Gly及LTLR2基因 Arg753Glu及Arg677Trp多态性在中国汉族人群中的分布,探讨其与炎症性肠病的相关性．方法:采用聚合酶链反应-限制性片段长度多态性方法,检测120例中国湖北汉族炎症性肠病患者与110例正常对照者TLR4 基因ASp299Gly及TLR2基因Arg753Glu及 Arg677Trp基因型,分析该基因多态性与炎症性肠病以及临床亚型的相关性．结果:炎症性肠病患者和健康对照者均未检测出TLR4基因ASp299Gly及TLR2基因 Arg753Glu及Arg677Trp突变型．结论:TLR4基因Asp299Gly及TLR2基因 Arg753Glu及Arg677Trp基因多态性与中国湖北汉族人群炎症性肠病的易感性无相关性．     

TLR2 and TLR4 polymorphisms influence mRNA and protein expression in colorectal cancer

AIM: To evaluate the effect of promoter region polymorphisms of toll-like receptor(TLR)2-196 to-174 del and TLR4-1607T/C(rs10759932) on m RNA and protein expression in tumor tissue and of TLR4+896A/G(rs4986790) on colorectal cancer(CRC) risk.METHODS: The TLR2-196 to-174 del polymorphism was investigated using allele-specific polymerase chain reaction(PCR) and the TLR4-1607T/C and TLR4+896A/G by PCR-restriction fragment length p o l y m o r p h i s m( R F L P). W e g e n o t y p e d 4 3 4 D N A samples from 194 CRC patients and 240 healthy individuals. The m RNA relative quantification(RQ) was performed in 40 tumor tissue samples by quantitative PCR Taq Man assay, using specific probes for TLR2 and TLR4 genes, and ACTB and GAPDH reference geneswere used as endogenous controls. Protein expression was analyzed by immunohistochemistry with specific primary antibodies.RESULTS: No association was found for TLR4-1607T/C and TLR4+896A/G by three statistical models(logadditive, dominant and recessive). However, based on dominant and log-additive models, the polymorphic variant TLR2-196 to-174 del was associated with increased CRC risk [dominant: odds ratio(OR) = 1.72, 95%CI: 1.03-2.89; P = 0.038 and log-additive: OR =1.59, 95%CI: 1.02-2.48; P = 0.039]. TLR2 m RNA expression was increased in tumor tissue(RQ = 2.36) when compared to adjacent normal tissue(RQ = 1; P 0.0001), whereas the TLR4 m RNA showed a basal expression(RQ = 0.74 vs RQ = 1, P = 0.452). Immunohistochemistry analysis of TLR2 and TLR4 protein expression was concordant with the findings of m RNA expression. In addition, the TLR2-196 to-174 del variant carriers showed m RNA relative expression 2.19 times higher than wild-genotype carriers. The TLR2 protein expression was also higher for the TLR2-196 to-174 del variant carriers [117 ± 10 arbitrary unit(a.u.) vs 95 ± 4 a.u., P = 0.03]. However, for the TLR4-1607T/C polymorphism no significant difference was found for both m RNA(P = 0.56) and protein expression(P = 0.26).CONCLUSION: Our findings suggest that TLR2-196 to-174 del polymorphism increases TLR2 m RNA expression and is associated with higher CRC risk, indicating an important role in CRC genetic susceptibility.     

Different Doses of Lipopolysaccharides Regulate the Lung Inflammation of Asthmatic Mice via TLR4 Pathway in Alveolar Macrophages

Allergic asthma is a complicated genetic disorder caused by interaction of the acquired and innate immune responses. Acquired immune responses to protein antigens could induce type 2 T lymphocyte-driven responses and result in atopic asthma. Recent studies demonstrated that endotoxin, LPS and air pollution-induced innate immunity induce asthma through Toll-like receptors (TLR). However, the definite mechanism of LPS-induced asthma is still not known. Here, we investigated the effects of different doses of LPS in a mouse model of allergic asthma to define the molecular mechanism of LPS-induced asthma. We found that low doses of LPS in OVA induced significant inflammatory infiltration in lung tissue of asthmatic mice. Histologic studies demonstrated that lungs of these asthmatic mice were characterized by the recruitment of both eosinophils and neutrophils, increased airway mucus secretion and the elevated levels of Th2 cytokines. A high dose of LPS in OVA can induce a Th1 associated response, histologically characterized by neutrophil recruitment, the absence of airway mucus secretion and an increase of IFN-γ production. Regardless of high or low dose of LPS, TLR4 in alveolar macrophages (AM) was up-regulated in lungs of asthmatic mice. Our data demonstrated that the dose of LPS exposure determines the type of inflammatory response and a low dose of LPS together with OVA augments the antigen-induced lung inflammation in asthma. This study demonstrates that the TLR4 signaling pathway plays a vital role in the development of asthma and indicates the tight connection between endotoxin exposure and asthma prevalence in the clinic.     

慢性乙型肝炎患者外周血单核细胞Toll样受体2和4的变化

目的探讨慢性乙型肝炎和慢性重型乙型肝炎患者外周血单核细胞Toll样受体2 （TLR2）和Toll样受体4（TLR4）的变化情况及其意义。方法用流式细胞仪检测正常对照、慢性乙型肝炎患者和慢性重型乙型肝炎患者外周血单核细胞表面TLR2和TLR4的表达,ELISA法检测上述患者血清TNFα的水平。Student-t检验比较3组间TLR2、TLR4和TNFα表达的差异;3组病例TLR2、TLR4的表达水平间及它们与血清TNFα水平间的相关性分析采用线性相关分析。结果正常对照组（30例）、慢性乙型肝炎组（31例）和慢性重型乙型肝炎组（30例）外周血单核细胞TLR2和TLR4的平均免疫荧光强度（MFI）分别为21．5±2．7、39．0±4．1．47．7±21．4和2．3±1．1、3．7±2．3、6．9±4．1;外周血清TNFα（ng／L）水平分别为53．8±38．1、164．3±89．9、359．8±1 40．0,自正常对照组到慢性乙型肝炎组及慢性重型乙型肝炎组外周血单核细胞TLR2、TLR4表达强度和外周血清TNFα水平均依次显著升高;经线性相关分析研究发现,外周血单核细胞TLR2、TLR4表达水平和血清TNFα表达水平间均呈现显著的正相关性。结论TLR2和TLR4可能与慢性乙型肝炎及慢性重型乙型肝炎的发病有关。     

Toll样受体4与非酒精性脂肪性肝病的关系

非酒精性脂肪性肝病(NAFLD)常发生于具有代谢综合征特点的患者,主要临床病理表现为肝实质细胞脂肪变性和脂肪贮积,可进展为肝硬化,最终发生肝衰竭。Toll样受体(TLR)4信号途径参与NAFLD的发生发展。目前,TLR4在NAFLD发病中所起到的作用引起临床医师高度重视。简述了TLR4的结构、TLR4信号途径的构成、表达调节方式、功能及其与NAFLD的关系,认为未来TLR4信号途径必然成为治疗NAFLD的重要研究领域。     

右美托咪定联合Tol样受体4抑制剂对缺氧复氧心肌细胞凋亡和炎症反应的影响及其机制

目的探讨右美托咪定(DEX)联合Toll样受体4(TLR4)抑制剂TAK-242对缺氧复氧(H/R)心肌细胞凋亡和炎症反应的影响及其机制。方法心肌细胞H9C2分为对照(Con)组、H/R组(H/R损伤)、DEX组(1.0μmol/L DEX,再行H/R损伤)、TAK-242组(30μmol/L TAK-242,再行H/R损伤)和DEX+TAK-242组(30μmol/L TAK-242及1.0μmol/L DEX,再行H/R损伤处理)。各组细胞复氧培养6 h后,采用MTT法、流式细胞仪、试剂盒、酶联免疫吸附法检测细胞增殖、凋亡、乳酸脱氢酶(LDH)释放率、白介素1β(IL-1β)和肿瘤坏死因子-α(TNF-α)含量,Western blot检测B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、裂解的caspase-3(cleaved caspase-3)、TLR4和核因子B p65(NF-κB p65)蛋白表达。结果五组细胞凋亡率、LDH释放率、Bax、Bcl-2、cleaved caspase-3、TLR4、NF-κB p65蛋白表达水平、IL-1β、TNF-α含量比较差异均有统计学意义(F=316.938、330.004、839.933、169.750、378.365、476.535、298.527、99.219、293.498,P<0.05)。与Con组相比,H/R组细胞的凋亡率、LDH释放率、Bax、cleaved caspase-3、TLR4和NF-κB p65蛋白表达水平、细胞上清液中IL-1β和TNF-α含量均明显升高(均P<0.05),Bcl-2蛋白表达水平明显降低(均P<0.05)。与H/R组相比,DEX组、TAK-242组和DEX+TAK-242组的细胞凋亡率、LDH释放率、Bax蛋白、cleaved caspase-3、TLR4和NF-κB p65蛋白、IL-1β、TNF-α的表达水平均明显降低,Bcl-2蛋白表达水平明显升高(均P<0.05)。与DEX组、TAK-242组相比,DEX+TAK-242组的细胞凋亡率、LDH释放率、Bax、cleaved caspase-3、TLR4和NF-κB p65蛋白表达、IL-1β、TNF-α表达水平更低,Bcl-2蛋白的表达更高(均P<0.05)。结论DEX和TAK-242联合可协同抑制H/R引起的心肌细胞凋亡和炎症反应,其作用机制可能与协同抑制TLR4/NF-κB通路有关。     

蓝莓下调Toll样受体4表达减轻肝损伤的作用机制

目的研究蓝莓对Toll样受体4(TLR4)表达的调节,进一步探讨其在急性肝损伤中的作用。方法 48只SD大鼠随机分为空白对照组(对照组)、CCl4急性肝损伤组(模型组)、蓝莓低剂量组、蓝莓高剂量组和胸腺肽组。蓝莓低剂量组、蓝莓高剂量组造模开始前先分别给予蓝莓果汁(0.5 ml/100 g、1.0 ml/100 g)灌胃7 d。实验结束时测定肝脏指数、血清ALT、AST含量,WesternBlot检测肝组织TLR4蛋白表达量。结果模型组大鼠的肝脏指数、血清ALT、AST含量(0.047 30±0.002 85、4536.00±535.39,7959.33±108.89)高于对照组(0.031 90±0.002 87,48.00±5.57,137.00±12.53)(P<0.05);蓝莓低、高剂量组上述指标(0.040 80±0.003 77,0.040 80±0.003 81;2138.00±980.32,4049.67±466.01;1247.00±609.00,2063.33±1357.32)均低于模型组,模型组肝组织TLR4蛋白表达量(1.392±0.204)显著高于对照组(0.781±0.132)(P<0.05)。蓝莓低剂量组、高剂量组和胸腺肽组大鼠肝组织TLR4蛋白表达量(1.273±0.203,1.110±0.168,0.943±0.161)低于模型组(1.392±0.204)(P<0.05)。结论蓝莓可能通过下调肝组织中TLR4的表达,起到减轻急性肝损伤的作用。     

丙型肝炎病毒NS5A在肝癌细胞对TLR4表达的影响

目的探讨丙型肝炎病毒非结构蛋白质5A（HCV NS5A）对Toll样受体4（TLR4）表达的影响。方法用含HCV NS5A基因的表达质粒pcNS5A瞬时转染QSG7701细胞,采用免疫细胞化学技术检测HCV NS5A及TLR4蛋白质的表达;采用逆转录聚合酶链反应观察HCV NS5A和TLR4的mRNA表达。结果转染pcNS5A质粒细胞内有HCV NS5A蛋白的表达。转染pcNS5A组TLR4 mRNA及蛋白质表达均较转染pRc/CMV和未转染质粒组明显增强,而后两组细胞TLR4 mRNA和蛋白质表达相似。结论 HCV NS5A可上调TLR4 mRNA和蛋白的表达。     

Obstructive jaundice increases sensitivity to lipopolysaccharide via TLR4 upregulation: possible involvement in gut-derived hepatocyte growth factor-protection of hepatocytes

BACKGROUND: Patients with obstructive jaundice are prone to sepsis after biliary tract surgery. The present study was designed to determine the effect of biliary obstruction on cytokine responses to lipopolysaccharide (LPS). METHODS: Wister rats were allocated into two groups; the BDL group underwent bile duct ligation, followed 2 weeks later by administration of LPS into the duodenum. The control group underwent sham operation, and similarly received enteral LPS. Specimens were collected serially, and applied for the assays. RESULTS: Serum aspartate aminotransferase and alanine aminotransferase levels were significantly increased in BDL rats. High tumor necrosis factor alpha (TNF-alpha) and interleukin (IL)-6 levels in peripheral blood were observed 2 h after LPS administration in BDL rats. In contrast, no increases in both cytokines were noted in peripheral and portal blood in control rats. Baseline HGF levels in portal and peripheral blood in BDL rats were significantly higher than in control rats. LPS significantly increased hepatocyte growth factor (HGF) levels in portal blood and decreased in peripheral blood in BDL rats, but not in control rats. Immunohistochemical analysis revealed that BDL increased expressions of Toll-like receptor (TLR)4, CD14 and CD68 both in the small intestine and liver. Both TLR4 and CD14 mRNAs were upregulated in the small intestine and liver after LPS administration in BDL rats. CONCLUSION: Obstructive jaundice and LPS stimulation induced TLR4 upregulation both in the liver and small intestine, which led to increased TNF-alpha and IL-6 production in liver and HGF production in the small intestine. The upregulation of TLR4 may lead to pathological and host defense reactions in obstructive jaundice complicated with Gram-negative bacterial infection.     

Toll样受体4在胰腺癌组织中的表达及其与肿瘤血管生成的关系

目的 探讨Toll样受体4(TLR4)在胰腺癌组织中的表达及其与肿瘤血管生成的关系.方法 采用免疫组化SP法检测62例经病理证实的胰腺癌手术切除标本及35例癌旁正常胰腺组织中TLR4蛋白表达,采用CD31抗体标记微血管内皮细胞,计算微血管密度(MVD).分析TLR4蛋白表达与胰腺癌临床病理特征以及MVD的相关性.结果 胰腺癌组织TLR4蛋白阳性表达率和MVD分别为74.2％ (46/62)和47.3±13.5,均显著高于正常胰腺组织的17.1％ (6/35)和12.6±4.8(P值均＜0.01).有淋巴结转移的胰腺癌组织中TLR4蛋白阳性表达率为83.8％,显著高于无淋巴结转移的60.0％(P=0.036);TNM分期为Ⅲ+Ⅳ期的胰腺癌组织中TLR4蛋白阳性表达率为85.3％,显著高于Ⅰ+Ⅱ期的60.7％(P =0.028).MVD与肿瘤的大小、淋巴结转移及TNM分期密切相关(P值分别为0.008、0.036、0.010).胰腺癌TLR4蛋白表达与MVD呈显著正相关(Υ=0.534,P＜0.01).结论 TLR4参与胰腺癌的发生、发展,其机制可能与促进肿瘤血管生成有关.     

蓝莓对免疫性肝纤维化大鼠肝组织TLR4,TLR9表达的影响

目的:探讨蓝莓抗氧化作用对猪血清免疫性肝纤维化大鼠肝组织Toll样受体4(toll-like receptor 4,TLR4)和Toll样受体9(TLR9)蛋白表达的影响.方法:60只♂Wistar大鼠随机分为正常对照组(A组)、模型组(B组)、蓝莓低、中、高剂量防治组(C、D、E组)、复方鳖甲软肝片防治组(F组).除A组外,其余各组均腹腔注射猪血清制备大鼠肝纤维化模型.C、D、E、F组在造模同时分别给予蓝莓原浆或复方鳖甲软肝片灌胃,1次/d,共12wk.第12周末处死大鼠,计算大鼠肝脏指数,行肝脏病理组织学检查,测定各组大鼠血清ALT和AST水平、肝组织匀浆超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)及羟脯氨酸(hydroxyproline,Hyp)含量,免疫印迹法(western blot,WB)和免疫组织化学法检测TLR4和TLR9蛋白质在肝脏组织中的表达.结果:各组大鼠血清ALT、AST水平无明显差异(F=4.274,0.917,P>0.05);与B组比较,D,E组肝组织匀浆SOD活性、GSH含量明显升高(F=6.662,61.472,P<0.05);肝脏指数、MDA及Hyp含量明显降低(F=3.972,42.755和41.265,P<0.05);肝纤维化程度明显减轻(F=63.339,P<0.05),胶原表达减少;E组大鼠肝组织TLR4和TLR9蛋白质表达明显减少(F=6.932和7.534,P<0.05).结论:TLR4和TLR9蛋白质表达增加可能与免疫性肝纤维化的发生发展有关.蓝莓对大鼠免疫性肝纤维化有一定的预防作用.