Effects of ω-3 fatty acids on toll-like receptor 4 and nuclear factor-κB p56 in lungs of rats with severe acute pancreatitis

AIM To determine the effects of ω-3 fatty acids(ω-3FA) on the toll-like receptor 4(TLR4)/nuclear factor κB p56(NF-κBp56) signal pathway in the lungs of rats with severe acute pancreatitis(SAP).METHODS A total of 56 Sprague-Dawley rats were randomly divided into 4 groups: control group, SAP-saline group, SAP-soybean oil group and SAP-ω-3FA group. SAP was induced by the retrograde infusion of sodium taurocholate into the pancreatic duct. The expression of TLR4 and NF-κBp56 in the lungs was evaluated by immunohistochemistry and Western blot analysis. The levels of inflammatory cytokines interleukin-6 and tumor necrosis factor-alpha in the lungs were measured by enzyme-linked immunosorbent assay. RESULTS The expression of TLR4 and NF-κBp56 in lungs and of inflammatory cytokines in serum significantly increased in the SAP group compared with the control group(P < 0.05), but was significantly decreased in the ω-3FA group compared with the soybean oil group at 12 and 24 h(P < 0.05).CONCLUSION During the initial stage of SAP, ω-3FA can efficiently lower the inflammatory response and reduce lung injury by triggering the TLR4/NF-κBp56 signal pathway.

5-HT₄Receptor Agonists for the Treatment of Alzheimer’s Dsease

Alzheimer’s disease (AD) is a progressive neurological disorder primarily affecting new memory formation as well as retrieval of previously acquired memories. According to World Health Organization, current global population suffering from cognitive impairment is estimated to 37 million. The number is projected to double in next one and half decade. Half of the population afflicted with dementia is represented by AD patients. Current therapies, which provide marginal symptomatic relief to AD patients, are effective only in half of the patient population. In depth understanding of the molecular mechanism of the disease is urgently required to develop more effective therapies. Therapies in clinical development may either offer symptomatic relief to patients or provide pure disease modifications, thus limiting benefit to patients. 5-HT4 receptor agonists offer an attractive option for the treatment of AD patients. Activation of 5- HT4 receptor under preclinical conditions is demonstrated to improve neurotransmission and enhance the release of acetylcholine resulting in the memory formation. In various cell based and animal models, partial 5-HT4 receptor agonists are demonstrated to promote the release of soluble amyloid precursor protein alpha and block the release of amyloid beta peptide offering suitable candidates as disease modification agents. Remarkably, 5-HT4 receptor agonists are also reported to induce neurogenesis in hippocampus as well as enteric system through the activation of cyclic AMP response element binding protein in rodents. Taken together, 5-HT4 agonists address all major facets of Alzheimer’s disease and may provide therapeutic potential for other neurological disorders.

脂多糖对2型糖尿病肾病患者单核细胞TLR4水平及NF-κB,Notch1表达的影响

目的:探讨2型糖尿病肾病(diabetic nephropathy,DN)患者外周血单核细胞Toll样受体4(toll-like receptor,TLR4)的表达及脂多糖(lipopolysaccharide,LPS)对其的影响,以了解炎症性免疫反应在DN中的可能作用机制。方法:收集30例2型DN尿毒症患者,10例早期2型DN患者和20例健康志愿者。采用外周血流式细胞术检测单核细胞TLR4的表达,并分离其外周血单个核细胞(PBMC),用LPS干预24 h,收集PBMC和上清液。分别采用Western印迹检测PBMC内TLR4,NF-κB p65和Notch1蛋白表达;免疫荧光检测NF-κB p65蛋白表达。ELISA法检测外周血血清及LPS干预后细胞上清液IL-6浓度。免疫比浊法检测血清CRP水平。结果:2型DN患者外周血单核细胞的TLR4荧光强度值、IL-6和CRP水平均高于正常对照组[TLR4荧光强度值:2型DN尿毒症患者2.8±0.9、早期2型DN患者3.4±0.7、正常对照组1.6±0.7;IL-6:2型DN尿毒症患者(84.8±20.7)pg/mL、早期2型DN患者(63.20±14.4)pg/mL、正常对照组(11.0±2.0)pg/mL;CRP:2型DN尿毒症患者(5.4±2.8)mg/L;早期2型DN患者(3.7±1.7)mg/L、正常对照组(1.7±0.7)mg/L,均P<0.01],LPS干预后2型DN患者PBMC的TLR4,NF-κB P65蛋白表达和IL-6水平明显高于正常对照组(P<0.05),早期2型DN患者PBMC以上指标水平均高于DN尿毒症患者,且2型DN患者NF-κB p65的表达存在核转移现象,但与正常对照组比较Notch1蛋白表达水平差异无统计学意义(P>0.05)。结论:DN患者体内可能存在单核细胞功能紊乱,这种功能紊乱可能与TLR4/NF-κB信号通路的激活有关,与Notch1信号通路无关。

血清TLR4、HMGB1、IL-6与非酒精性脂肪性肝病患者动脉粥样硬化的相关性

目的:探讨血清Toll样受体4(TLR4)、高迁移率族蛋白B1(HMGB1)、白细胞介素-6(IL-6)与非酒精性脂肪性肝病(NAFLD)患者动脉粥样硬化(AS)的相关性。方法:2017年5月至2019年1月收治NAFLD患者116例,根据是否合并AS分为AS组(n=61)和NAS组(n=55)。比较两组患者的基本资料、血清TLR4、HMGB1、IL-6水平,并进行Pearson相关性分析和Logistic回归分析。结果:两组患者性别比例、年龄、吸烟史、BMI、DBP、FPG、HDL-C均差异无统计学意义(P>0.05)。AS组患者的病程、SBP、TC、TG、LDL-C、颈动脉内膜中层厚度(cIMT)、TLR4、HMGB1、IL-6均显著高于NAS组(P<0.05)。经过Pearson相关性分析,SBP、TC、TG、LDL-C、TLR4、HMGB1、IL-6与AS组患者cIMT均呈正相关(P<0.05)。经Logistic回归分析,SBP、LDL-C、TLR4、HMGB1、IL-6是AS组患者cIMT增加的危险因素。结论:TLR4、HMGB1、IL-6与NAFLD患者发生AS有关,因此应重视对NAFLD患者血压、血脂及相关炎性因子的监测和干预。

5-氮杂胞苷对胃癌裸鼠移植瘤死亡受体DR4和DR5表达的影响

【目的】研究5-氮杂胞苷(5-Aza-CdR)对胃癌裸鼠移植瘤中死亡受体4(DR4)和死亡受体5(DR5)表达的影响。【方法】利用RT-PCR方法检测5-Aza-CdR处理后的胃癌裸鼠移植瘤内DR4和DR5的RNA表达水平、用MS-PCR检测5-Aza-CdR处理后的胃癌裸鼠抑制瘤内DR4和DR5启动子区甲基化水平。【结果】5-Aza-CdR能够抑制胃癌裸鼠移植瘤的生长。5-Aza-CdR能够逆转胃癌裸鼠移植瘤中DR4和DR5的启动子甲基化水平,并上调DR4和DR5的表达水平。【结论】去甲基化药物5-Aza-CdR能够抑制裸鼠胃癌移植瘤生长,提示该药物有治疗作用且能够诱导DR4和DR5的启动子区去甲基化从而使DR4和DR5的表达水平升高。

基质细胞衍生因子-1及其受体4与动脉粥样硬化的研究进展

动脉粥样硬化是一种由遗传及环境等多种综合因素所致的疾病,最近研究发现,基质细胞衍生因子-1（stromal cell-derived factor-1,SDF-1）与其受体4（CXCR chemokine receptor 4,CXCR4）结合,通过活化和介导炎症细胞进入动脉粥样斑块内, 促进动脉粥样硬化和斑块不稳定.因而,以SDF-1/CXCR4 轴为靶点治疗动脉粥样硬化相关疾病日益受到人们的重视.现就SDF-1/CXCR4生物轴与动脉粥样硬化方面的研究综述如下.

Toll样受体4在慢性鼻-鼻窦炎伴鼻息肉组织中的研究进展

慢性鼻-鼻窦炎伴鼻息肉（chronic rhinosinusitis with nasal polyps）为耳鼻咽喉-头颈外科常见疾病,是以上皮细胞损害和组织水肿为特征的炎症性病变,其具体的发病机制尚存在许多不明之处.有学者认为鼻息肉的病因复杂,可能是由多个基因及多种环境因素共同作用的结果[1].病原体感染也是重要的致病因素,Toll样受体（toll-like receptor,TLR）属于模式识别受体能识别病原微生物细胞壁成分,其中TLR-4作为Toll样受体的重要组成部分,已有研究显示在鼻息肉组织中的表达明显高于健康对照组[2],说明其对鼻息肉的病因及发展有重要意义,为此本文对TLR-4在慢性鼻-鼻窦炎伴鼻息肉的相关研究进行综述.

Metformin regulates inflammation and fibrosis in diabetic kidney disease through TNC/TLR4/NF-κB/miR-155-5p inflammatory loop

BACKGROUND Type 2 diabetes mellitus(T2DM)is significantly increasing worldwide,and the incidence of its complications is also on the rise.One of the main complications of T2DM is diabetic kidney disease(DKD).The glomerular filtration rate(GFR)and urinary albumin creatinine ratio(UACR)increase in the early stage.As the disease progresses,UACR continue to rise and GFR begins to decline until endstage renal disease appears.At the same time,DKD will also increase the incidence and mortality of cardiovascular and cerebrovascular diseases.At present,the pathogenesis of DKD is not very clear.Therefore,exploration of the pathogenesis of DKD to find a treatment approach,so as to delay the development of DKD,is essential to improve the prognosis of DKD.AIM To detect the expression of tenascin-C(TNC)in the serum of T2DM patients,observe the content of TNC in the glomerulus of DKD rats,and detect the expression of TNC on inflammatory and fibrotic factors in rat mesangial cells(RMCs)cultured under high glucose condition,in order to explore the specific molecular mechanism of TNC in DKD and bring a new direction for the treatment of DKD.METHODS The expression level of TNC in the serum of diabetic patients was detected by enzyme-linked immunosorbent assay(ELISA),the protein expression level of TNC in the glomerular area of DKD rats was detected by immunohistochemistry,and the expression level of TNC in the rat serum was detected by ELISA.Rat glomerular mesangial cells were cultured.Following high glucose stimulation,the expression levels of related proteins and mRNA were detected by Western blot and polymerase chain reaction,respectively.RESULTS ELISA results revealed an increase in the serum TNC level in patients with T2DM.Increasing UACR and hypertension significantly increased the expression of TNC(P<0.05).TNC expression was positively correlated with glycosylated haemoglobin(HbA1c)level,body mass index,systolic blood pressure,and UACR(P<0.05).Immunohistochemical staining showed that TNC expression in the glomeruli of rats with streptozotocin-induced diabetes was significantly increased compared with normal controls(P<0.05).Compared with normal rats,serum level of TNC in diabetic rats was significantly increased(P<0.05),which was positively correlated with urea nitrogen and urinary creatinine(P<0.05).The levels of TNC,Toll-like receptor-4(TLR4),phosphorylated nuclear factor-κB p65 protein(Ser536)(p-NF-κB p65),and miR-155-5p were increased in RMCs treated with high glucose(P<0.05).The level of TNC protein peaked 24 h after high glucose stimulation(P<0.05).After TNC knockdown,the levels of TLR4,p-NF-κB p65,miR-155-5p,connective tissue growth factor(CTGF),and fibronectin(FN)were decreased,revealing that TNC regulated miR-155-5p expression through the TLR4/NF-κB p65 pathway,thereby regulating inflammation(NF-κB p65)and fibrosis(CTGF and FN)in individuals with DKD.In addition,metformin treatment may relive the processes of inflammation and fibrosis in individuals with DKD by reducing the levels of the TNC,p-NF-κB p65,CTGF,and FN proteins.CONCLUSION TNC can promote the occurrence and development of DKD.Interfering with the TNC/TLR4/NF-κB p65/miR-155-5p pathway may become a new target for DKD treatment.

Effect of ω-3 Polyunsaturated Fatty Acid on Toll-like Receptors in Patients with Severe Multiple Trauma

This study examined the effects of ω-3 polyunsaturated fatty acid（ω-3PUFA） on the expression of toll-like receptor 2（TLR2）,toll-like receptor 4（TLR4） and some related inflammatory factors in peripheral blood mononuclear cells（PBMCs） of patients with early-stage severe multiple trauma.Thirty-two patients who were admitted to the Department of Traumatic Surgery,Tongji Hospital（Wuhan,China） between May 2010 and November 2010,and diagnosed as having severe multiple trauma with a injury severity score（ISS） no less than 16,were enrolled in the study and divided into two groups at random（n=16 in each）：ω-3PUFA group and control group in which routine parenteral nutrition supplemented with ω-3PUFA or not was administered to the patients in two groups for consecutive 7 days.Peripheral blood from these patients was collected within 2 h of admission（day 0）,and 1,3,5 and 7 days after the nutritional support.PBMCs were isolated and used for detection of the mRNA and protein expression of TLR2 and TLR4 by using real-time PCR and flow cytometry respectively,the levels of NF-κB by quantum dots-based immunofluorescence assay,the levels of TNF-α,IL-2,IL-6 and COX-2 by ELISA,respectively.The results showed that the mRNA and protein expression of TLR2 and TLR4 in PBMCs was significantly lower in ω-3PUFA group than in control group 5 and 7 days after nutrition support（both P0.05）.The levels of TNF-α,IL-2,IL-6 and COX-2 were found to be substantially decreased in PBMCs in ω-3PUFA group as compared with control group at 5th and 7th day（P0.05 for all）.It was concluded that ω-3PUFA can remarkably decrease the expression of TLR2,TLR4 and some related inflammatory factors in NF-κB signaling pathway in PBMCs of patients with severe multiple trauma,which suggests that ω-3PUFA may suppress the excessive inflammatory response meditated by the TLRs/NF-κB signaling pathway.

Endozepine-4 levels are increased in hepatic coma

AIM:To evaluate the serum levels of endozepine-4,their relation with ammonia serum levels,the grading of coma and the severity of cirrhosis,in patients with hepatic coma. METHODS:In this study we included 20 subjects with Hepatic coma,20 subjects with minimal hepatic encephalopathy(MHE) and 20 subjects control. All subjects underwent blood analysis,Child Pugh and Model for End- stage liver disease(MELD) assessment,endozepine-4 analysis. RESULTS:Subjects with hepatic coma showed significant difference in endozepine-4(P < 0.001) and NH3 levels(P < 0.001) compared both to MHE and controls patients. Between NH3 and endozepine-4 we observed a significant correlation(P = 0.009; Pearson correlation 0.570). There was a significant correlation between endozepine-4 and MELD(P = 0.017; Pearsoncorrelation = 0.529). In our study blood ammonia concentration was noted to be raised in patients with hepatic coma,with the highest ammonia levels being found in those who were comatose. We also found a high correlation between endozepine-4 and ammonia(P < 0.001). In patients with grade Ⅳ hepatic coma,endozepine levels were significantly higher compared to other groups. CONCLUSION:This study suggests that an increased level of endozepine in subjects with higher levels of MELD was observed. In conclusion,data concerning involvement of the GABA-ergic system in HE coma could be explained by stage-specific alterations.

EP4 agonist alleviates indomethacin-induced gastric lesions and promotes chronic gastric ulcer healing

AIM： To investigate EP4-selective agonist effect on indomethacin-induced gastric lesions and on the spontaneous healing of chronic gastric ulcers. METHODS： In a mouse model of gastric bleeding with high dose of indomethacin （20 mg/kg）, an EP4-selective agonist was administered orally. Stomach lesions and gastric mucous regeneration were monitored. In a mouse model of chronic gastric ulcer induced by acetic acid, EP4 agonist effect on the healing of chronic gastric ulcer was evaluated in the presence or absence of low dose indomethadn （3 mg/kg）. In cultured human gastric mucous cells, EP4 agonist effect on indomethacin- induced apoptosis was assessed by flow cytometry. RESULTS： The EP4-selective agonist reduced high dose indomethacin-induced acute hemorrhagic damage and promoted mucous epithelial regeneration. Low-dose indomethacin aggravated ulcer bleeding and inflammation, and delayed the healing of the established chronic gastric ulcer. The EP4 agonist, when applied locally, not only offset indomethacin-induced gastric bleeding and inflammation, but also accelerated ulcer healing. In the absence of indomethacin, the EP4 agonist even accelerated chronic gastric ulcer healing and suppressed inflammatory cell infiltration in the granulation tissue. In vitro, the EP4 agonist protected human gastric mucous cells from indomethacin-induced apoptosis.CONCLUSION： EP4-selective agonist may prevent indomethacin-induced gastric lesions and promote healing of existing and i ulcers, via promoting mucous epithelial cells. proliferation and survival of mucous epithelial cells.

青蒿琥酯对小鼠巨噬细胞RAW264.7中TLR4介导的炎症通路的抑制作用

目的观察青蒿琥酯(artesunate,AS)对脂多糖(lipopolysaccharide,LPS)刺激小鼠RAW264.7细胞Toll样受体4(Toll-like receptor 4,TLR4)介导炎症通路活化的影响,以探讨AS的抗炎作用机制。方法采用免疫荧光观察胞内TLR4表达和分布;免疫印迹检测TLR4及下游炎症通路关键分子的表达和活化;酶联免疫吸附法检测细胞培养上清中TNF-α、IL-6浓度。结果 AS对LPS诱导的TLR4表达及其在细胞中的聚集均有抑制作用;AS同时抑制TLR4衔接蛋白髓分化因子88和含TIR结构域干扰素诱导衔接蛋白表达,也抑制依赖于二者活化的肿瘤坏死因子受体相关因子6表达;对下游MAPK通路,AS抑制p38表达和磷酸化、JNK磷酸化,但对ERK1/2无显著影响;对下游NF-κB通路,AS下调抑制性-κBα(inhibitory-κBα,IκBα)的磷酸化,进而减少NF-κB亚基p50和p65活化入核的数量;最后,AS能够抑制LPS诱导的TNF-α和IL-6释放。结论 AS通过抑制LPS诱导的TLR4通路活化,减少致炎细胞因子的释放,从而发挥其抗炎作用。

Toll样受体2(TLR2)和TLR4在沙眼衣原体生殖道感染介导的免疫应答

目的 探讨Toll样受体2（TLR2）和TLR4在小鼠沙眼衣原体生殖道感染中的作用。方法用1×104包涵体形成单位的MoPn沙眼衣原体小鼠肺炎株经生殖道感染野生型（WT）小鼠（n=11）、 TLR2基因缺陷 （TLR2-/-）小鼠（n=14）和TLR4-/-小鼠（n=11）, 复制生殖道沙眼衣原体感染模型。在感染后第3、7、10、17、24、31、38、45天取阴道棉拭子, 免疫荧光法检测衣原体包涵体数量, ELISA检测白细胞介素1α（IL-1α）、IL-6和巨噬细胞炎性蛋白2（MIP-2）水平。感染后第70天, 采用免疫荧光法分析血清抗体类型和效价; 用MoPn沙眼衣原体感染无菌分离的腹腔巨噬细胞, 培养24 h, ELISA测定上清液中IL-1α、IL-6和MIP-2含量; 体外用紫外线灭活的衣原体抗原刺激无菌分离的脾细胞, 培养72 h, ELISA测定上清液中γ干扰素（IFN-γ）、IL-17、IL-4和IL-5水平。结果 与WT小鼠相比, TLR2-/-或TLR4-/-小鼠在各时间点的生殖道带菌量无差异、且带菌持续时间相同, 感染后第38天, 所有小鼠均清除了下生殖道感染的衣原体。TLR2-/-小鼠巨噬细胞产生的IL-1α、IL-6和MIP-2水平均明显低于WT小鼠,而TLR4-/-小鼠巨噬细胞产生的3种炎症细胞因子水平均与WT小鼠无显著性差异; TLR2-/-小鼠棉拭子标本具有较低水平的炎症细胞因子。所有小鼠脾细胞均产生高水平的IFN-γ和IL-17、 较低水平的IL-4和IL-5、血清IgG2a/ IgG1比值均大于1, 但各组间无显著性差异。结论 TLR2而不是TLR4介导了沙眼衣原体生殖道感染的早期免疫应答, 但沙眼衣原体诱导的适应性免疫应答可能既不依赖于TLR4, 也不依赖于TLR2。

内毒素诱导犬急性肺损伤TOLL样受体4的变化

目的探讨TOLL样受体4(TLR4)mRNA基因表达在内毒素休克犬急性肺损伤变化的规律。方法选择健康成年杂种雄性犬16条,随机分为对照组和实验组(n=8),实验组经心静脉30 min注入内毒素(LPS)650μg/kg,诱导犬急性肺损伤模型。用RT-PCR分别检测各组肺组织TLR4的表达。结果实验组与对照组相比,成模后4 h TLR4mRNA在实验组表达上调,2组比较差异有显著意义(P<0.01)。结论内毒素休克犬急性肺损伤时TLR4mRNA的表达上调。

Synthesis and biological evaluation of ^(99m)Tc-HEDTA/HYNIC-MPP4 complex for 5-HT_(1A) receptor imaging

5-HT_(1A)receptor is associated with a variety of pathophysiology of neuropsychiatric disorders.Accordingly,we have synthesized a new 5-HT_(1A) receptor ligand(HYNIC-MPP4)and labeled it with^(99m)Tc using N-(2-hydroxyethyl)ethylenediaminetriacetic acid(HEDTA)as coligand.^(99m)Tc-HEDTA/HYNIC-MPP4 was prepared under pH 6 at room temperature.Biodistribution of^(99m)Tc-HEDTA/HYNIC-MPP4 in normal mice showed that this complex had moderate brain uptake(0.60%ID·g^(-1)at 2 min p.i.)and good retention.The hippocampus had the highest radioactivity uptake at 2 min p.i.(1.84%ID·g^(-1)).The ratio of Hipp/CB was 3.1 at 2 min p.i.and increased to 4.4 at 60 min p.i.After blocking with 8-hydroxy-2-(dipropylamino)tetralin,the uptake of hippocampus was decreased significantly from 1.84%ID·g^(-1) to 0.53%ID·g^(-1) at 2 min p.i.,while the cerebellum had no significant decrease.This^(99m)Tc complex could be a potent agent for 5-HT_(1A) receptor imaging.

TLR途径负性调节因子A20、IRF-4和TRAF4在川崎病免疫发病中的变化

目的 探讨Toll样受体（TLR）信号途径负性调控因子在川崎病（KD）免疫发病机制中的作用。方法 急性期KD患儿48例，正常同年龄对照组16例，感染性疾病对照组（ID）16例。采用逆转录．聚合酶链反应（RT．PcR）及荧光定量PCR检测外周血单核，巨噬细胞（MC）TLR4信号途径传导分子、负性调节因子及前炎症细胞因子mRNA的表达。结果 （1）急性期KD患儿TLR4、MD-2、MyD88、IRAK-4、TRAF6、TAK1、TAB1及TAB2mRNA表达明显高于正常同年龄对照组（P〈0．01）；（2）KD及ID患儿A20、IRF-4、TRAF4基因表达上调（P〈0．01）；KD患儿除．A20表达水平高于ID对照组外，IRF-4、TRAF4表达水平明显低于ID对照组（P〈0．01）；KD患儿前炎症细胞因子表达明显高于ID组（P〈0．01）；（3）经细菌脂多糖（LPS）刺激后，KD患儿及正常对照组A20、IRF-4及TRAF4表达明显增高（P〈0．01），但KD患儿3种负性调节因子基因表达显著低于正常对照组（P〈0．01）；（4）KD合并冠状动脉损伤组（KD-CAL^＋）MC负性调节因子IRF-4及TRAF4明显低于无冠状动脉损伤组（KD-CAL^-），A20表达则显著高于后者；KD-CAL^＋组前炎症细胞因子IL-1β、IL-6、TNF-α均高于KD-CAL^-组（P〈0．01）。结论 急性期KD患儿TLR信号途径负性调节因子相对表达不足可能与KD的免疫发病机制有关。

益生菌对恶唑酮诱导小鼠结肠炎结肠上皮Toll样受体4和β-防御素3表达的影响

近年来对炎症性肠病（inflamrnatory bowel disease，IBD）动物模型的研究显示，非致病常驻菌是启动肠道慢性炎的必要条件，而益生菌能有效减轻各种实验性结肠炎。本研究通过建立与人类溃疡性结肠炎（ulcerative colitis，UC）类似的恶唑酮诱导小鼠结肠炎模型，继以益生菌VSL#3治疗，研究结肠上皮Toll样受体4（Toll like receptor 4，TLR4）及其附属蛋白髓样分化蛋白2（myeloid differential protein 2,MD2）、

溶血磷脂酸对THP-1细胞Toll样受体4/核因子-κB信号通路的影响

目的观察溶血磷脂酸（lysophosphatidieacid，LPA）对人单核细胞株THP-1细胞Toll-样受体4（tolllikereceptor4，TLR4）／核因子-κB（nuclearfactorkappaB，NF-κB）信号通路的影响，探讨LPA致动脉粥样硬化的机制。方法以不同浓度水平LPA（0～10μM）刺激人THP-1细胞4h，或以LPA1μM处理THP-1细胞不同时间（0-8h），荧光定量RT—PCR法测定TLR4mRNA表达，Westernblot检测TLR4蛋白、核蛋白NF-κBp65表达变化。结果IJPA以剂量依赖和时间依赖的方式上调THP-1细胞TLR4基因和蛋白的表达，并同步诱导THP-1细胞NF-κBp65活化。结论LPA可显著上调THP-1细胞TLR4表达及促进NF-κB的活化，LPA致粥样硬化作用可能部分是由TLR4／NF-κB信号途径介导的。

坎地沙坦抑制内毒素诱导的VSMCs炎症因子释放的作用研究

目的：研究坎地沙坦对内毒素（LPS）诱导血管平滑肌细胞（VSMCs）炎症因子释放的影响，探讨 Toll 样受体4（TLR4）介导的信号通路在这一过程中的作用。方法原代培养大鼠 VSMCs ，四甲基偶氮唑蓝（M TT ）法测定不同浓度坎地沙坦对 VSMCs 活性的影响。将细胞分为5组：A 组（对照组）、B 组（LPS 干预组）、C 组（LPS ＋10－7 mol／L 坎地沙坦）、D 组（LPS ＋10－6 mol／L 坎地沙坦）和 E 组（LPS ＋10－5 mol／L 坎地沙坦）。实时定量 PCR 和 Western blot 测定各组细胞 TLR4、髓样分化因子88（Myd88） mRNA 和蛋白的表达、NF‐κB（p65）的核转位水平；ELISA 测定各组细胞上清液白细胞介素‐1β（IL‐1β）和肿瘤坏死因子‐α（TNF‐α）分泌水平；DCFH‐DA 氧化法测定细胞内活性氧（iROS）含量。使用 TLR4阻滞剂 TLR4抗体、NADPH 氧化酶阻滞剂二联苯基碘（DPI）、NF‐κB 阻滞剂 PDTC 或坎地沙坦与阻滞剂联用预处理细胞，ELISA 法测定 IL‐1β和 TNF‐α分泌水平。结果坎地沙坦在10－8～10－3 mol／L 浓度范围内对 VSMCs 活性没有显著影响。同对照组相比，坎地沙坦可以有效抑制 LPS 诱导VSMCs IL‐1β和 TNF‐α的释放，减少 TLR4、Myd88 mRNA 和蛋白的表达以及 iROS 的生成，抑制 NF‐κB（p65）的核转位，并具有剂量依赖性。 TLR4抗体、DPI 、PDTC 均能有效地抑制 LPS 诱导的炎症因子释放，坎地沙坦与阻滞剂联用显示出更强的抗炎作用。结论坎地沙坦能够减少 LPS 诱导的 VSMCs 炎症因子 IL‐1β和 TNF‐α的分泌，这一作用可能是通过抑制 TLR4／Myd88‐iROS‐NF‐κB 信号通路而实现的。

TLR4和NF-κB p65在子宫内膜样癌中的表达及临床意义

目的:观察和分析子宫内膜样癌组织中Toll样受体4(TLR4)和核转录因子-κB p65(NF-κB p65)的表达水平及临床意义。方法:选取子宫内膜组织标本155例作为研究材料,其中正常增生期子宫内膜组织标本42例、不典型增生内膜组织标本37例、子宫内膜样癌组织标本76例。应用常规免疫组化SP法对TLR4和NF-κB p65的表达水平进行检测和比较。结果:正常子宫内膜组织标本中的TLR4和NF-κB p65表达水平显著低于子宫内膜非典型增生组织标本(P<0.05),子宫内膜非典型增生组织标本中的TLR4和NF-κB p65表达水平显著低于子宫内膜样癌组织标本(P<0.05);组织学分级为Ⅰ级的子宫内膜样癌组织标本中的TLR4和NF-κB p65表达水平显著低于组织学分级为Ⅱ级的标本(P<0.05),组织学分级为Ⅱ级的子宫内膜样癌组织标本中的TLR4和NF-κB p65表达水平显著低于组织学分级为Ⅲ级的标本(P<0.05),肌层浸润深度<1/2或无浸润的子宫内膜样癌组织标本中的TLR4和NF-κB p65表达水平显著低于肌层浸润深度≥1/2或有淋巴转移的标本(P<0.05)。结论:子宫内膜样癌的发生和发展可能与TLR4和NF-κB p65的高水平表达具有相关性,子宫内膜样癌患者子宫内膜组织标本中的TLR4和NF-κB p65表达水平可能与肿瘤的组织学分级和扩散程度具有相关性,可用于预测患者的病情严重程度和预后,为制定术后治疗方案提供参考依据。