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.

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.

Oxygen-glucose deprivation of neurons transfected with toll-like receptor 3-siRNA Determination of an optimal transfection sequence

Toll-like receptor 3 protein expression has been shown to be upregulated during cerebral ische- mia/reperfusion injury in rats. In this study, rat primary cortical neurons were subjected to oxy- gen-glucose deprivation to simulate cerebral ischemia/reperfusion injury. Chemically synthesized small interfedng RNA （siRNA）-1280, -1724 and -418 specific to toll-like receptor 3 were transfected into oxygen-glucose deprived cortical neurons to suppress the upregulation of toll-like receptor 3 protein expression. Western blotting demonstrated that after transfection with siRNA, toll-like re- ceptor 3 protein expression reduced, especially in the toll-like receptor 3-1724 group. These results suggested that siRNA-1724 is an optimal sequence for inhibiting toll-like receptor 3 expression in cortical neurons following oxygen-glucose deprivation.

Correlation analysis of Toll-like receptor 3 content in peripheral blood with electroencephalogram parameters and neurotransmitter content in children with intractable epilepsy

Objective:To detect the content of Toll-like receptor 3 (TLR3) in peripheral blood of children with intractable epilepsy, explore the correlation between TLR3 content and EEG parameters, neurotransmitter contents.Methods:37 cases of Intractable epilepsy children in our hospital during September 2016to June 2018 were chosen as Intractable epilepsy group, 30 cases of healthy volunteers who underwent physical examination in our hospital were treated as Normal control group. The levels of TLR3, neurotransmitter [5-hydroxytryptamine (5-HT), dopamine (DA), epinephrine (E), norepinephrine (NE)] and electroencephalogram parameters [alpha, beta, delta, theta] in peripheral blood of two groups were compared. Pearson test was used to evaluate the correlation of TLR3 content in peripheral blood with EEG parameters and neurotransmitter content in children with intractable epilepsy.Results: Content of TLR3 in peripheral blood of Intractable epilepsy group was significantly higher than that of Normal control group;the alpha power and theta power of EEG parameters were lower than those of Normal control group;contents of neurotransmitters such as 5-HT, DA, E and NE were significantly lower than those of Normal control group (P<0.05). The correlation analysis showed that content of TLR3 in peripheral blood of children with intractable epilepsy was negatively correlated with levels of alpha and theta power of EEG, positively correlated with content of neurotransmitters such as 5-HT, DA, E and NE (P<0.05), but had no significant correlation was found with level of beta and delta power (P>0.05).Conclusion: The abnormal increase of TLR3 in peripheral blood of children with intractable epilepsy may be one of the direct causes of neurological impairment in children with intractable epilepsy.

Jianpi Gushen Huayu decoction ameliorated diabetic nephropathy through modulating metabolites in kidney,and inhibiting TLR4/NF-κB/NLRP3 and JNK/P38 pathways

BACKGROUND Jianpi Gushen Huayu Decoction(JPGS)has been used to clinically treat diabetic nephropathy(DN)for many years.However,the protective mechanism of JPGS in treating DN remains unclear.AIM To evaluate the therapeutic effects and the possible mechanism of JPGS on DN.METHODS We first evaluated the therapeutic potential of JPGS on a DN mouse model.We then investigated the effect of JPGS on the renal metabolite levels of DN mice using non-targeted metabolomics.Furthermore,we examined the effects of JPGS on c-Jun N-terminal kinase(JNK)/P38-mediated apoptosis and the inflammatory responses mediated by toll-like receptor 4(TLR4)/nuclear factor-kappa B(NF-κB)/NOD-like receptor family pyrin domain containing 3(NLRP3).RESULTS The ameliorative effects of JPGS on DN mice included the alleviation of renal injury and the control of inflammation and oxidative stress.Untargeted metabolomic analysis revealed that JPGS altered the metabolites of the kidneys in DN mice.A total of 51 differential metabolites were screened.Pathway analysis results indicated that nine pathways significantly changed between the control and model groups,while six pathways significantly altered between the model and JPGS groups.Pathways related to cysteine and methionine metabolism;alanine,tryptophan metabolism;aspartate and glutamate metabolism;and riboflavin metabolism were identified as the key pathways through which JPGS affects DN.Further experimental validation showed that JPGS treatment reduced the expression of TLR4/NF-κB/NLRP3 pathways and JNK/P38 pathway-mediated apoptosis related factors.CONCLUSION JPGS could markedly treat mice with streptozotocin(STZ)-induced DN,which is possibly related to the regulation of several metabolic pathways found in kidneys.Furthermore,JPGS could improve kidney inflammatory responses and ameliorate kidney injuries in DN mice via the TLR4/NF-κB/NLRP3 pathway and inhibit JNK/P38 pathwaymediated apoptosis in DN mice.

泥蚶TLR3基因的生物学特性及表达分析

为探究泥蚶Toll样受体3(TLR3)基因的生物学特性及其对缺氧和硫化物胁迫的响应,克隆了泥蚶TgTLR3核心片段序列,并运用实时荧光定量PCR技术检测了TgTLR3基因在血淋巴、鳃、肝胰腺、外套膜、闭壳肌、斧足等组织的分布情况以及在缺氧和硫化物胁迫下血淋巴中的表达模式。结果显示,泥蚶TgTLR3开放阅读框2634 bp,编码877个氨基酸。其中,49~593位为胞外富含亮氨酸重复序列,跨膜域为658~680位,720~865位为白细胞介素受体1结构域,蛋白质分子量为99.4 kD,等电点为7.76。氨基酸同源性比对发现,泥蚶TgTLR3氨基酸序列与毛蚶相似度高达80.05%。系统进化树显示,TgTLR3与毛蚶首先聚为一支,表明与毛蚶亲缘关系最近。TgTLR3在泥蚶各组织中均有表达,其中血淋巴表达量最高。缺氧胁迫48~96 h后,血淋巴中TgTLR3显著升高;添加硫化物后,TgTLR3在48 h显著低于缺氧组但在96 h仍显著高于对照。上述研究结果表明,TgTLR3基因在泥蚶应对缺氧和硫化物胁迫的免疫调控中可能发挥着重要作用。研究结果为探究软体动物TLR3的功能及其响应缺氧及硫化物胁迫的免疫机制提供了基础数据。

TLR3和IRF3在华支睾吸虫感染C3H／HeN小鼠中表达的初步分析

目的 建立华支睾吸虫感染小鼠模型,初步分析TLR3、IRF3基因在小鼠肝脏中的表达情况.方法 随机选用健康雌性C3H/HeN小鼠感染华支睾吸虫建立感染模型,分别在感染第0、1、7、14、28、56、84天颈椎脱臼处死小鼠,取肝脏组织进行病理学观察,收集粪便检查华支睾吸虫虫卵.同时,采用逆转录聚合酶链反应（RT-PCR）检测肝脏中Toll样受体（TLRs）中的TLR3和IRF3的mRNA的表达情况.结果 自感染后第20~30天检查出华支睾吸虫虫卵;肝脏病理学结果显示,与正常组小鼠相比,随着时间的增加,感染组小鼠肝脏汇管区由少量炎症细胞浸润、胆管上皮反应性增生发展到在该区域出现大量纤维组织增生伴有炎症细胞浸润,表明华支睾吸虫感染C3H/HeN小鼠模型成功建立.与正常组相比,感染组自感染后第1天起TLR3和IRF3的mRNA表达均升高（P〈0.01）,均在第84天开始下降.结论 华支睾吸虫感染C3H/HeN小鼠后可激活TLR3-TRIF途径,提示TLR3-TRIF途径可能在华支睾吸虫致病过程中起了一定的作用.

肝细胞肝癌间质微环境与TLR3表达的相关性及其对预后的影响

目的分析肝细胞肝癌(hepatocellular carcinoma,HCC)间质微环境与Toll样受体3(Toll-like receptor 3,TLR3)表达的相关性及其对预后的影响。方法利用已有专利技术制备人体HCC组织芯片,通过免疫组织化学检测HCC间质中各项指标及HCC细胞中TLR3表达,各因素相关分析和Kaplanmeier生存分析上述指标与TLR3的关系,及其与HCC预后的关系。结果 HCC间质微环境中浸润的T细胞(P=0.002)、Kupffer细胞(P=0.049)、自然杀伤(natural killer,NK)细胞(P=0.000)和树突状细胞(dendritic cells,DCs)(P=0.027)与TLR3表达呈正相关,而肥大细胞(P=0.000)、癌相关肌纤维母细胞(carcinoma-associated fibroblasts,CAFs)(P=0.000)和微血管密度(microvessel density,MVD)(P=0.000)与TLR3表达呈负相关;Kupffer细胞(P=0.013)和NK细胞(P=0.001)与预后呈正相关,而肥大细胞(P=0.008)、CAFs(P=0.000)和MVD(P=0.089)与预后呈负相关。结论间质微环境在HCC的演进过程中起着重要作用。

TLR3在poly(I:C)-LMW预处理后缺糖缺氧诱导的原代皮质神经元的表达意义

目的观察TRL3激动剂poly(I:C)-LMW预处理对原代皮质神经元Toll样受体3(TLR3)表达的影响,探讨TLR3在缺糖缺氧诱导的原代皮质神经元损伤中的作用。方法取体外培养7d的大鼠原代皮质神经元细胞,对细胞分别予以正常条件下培养(空白对照组);缺糖缺氧2h,复糖复氧24h(OGD组);TRL3激动剂预处理12h(激动剂组);TRL3激动剂预处理12h后缺糖缺氧2h,复糖复氧24h(激动剂+OGD组)。免疫荧光法观察各组细胞生长状态,分别以Western blot法、RT-PCR法测定TLR3蛋白及TLR3mRNA的表达情况。结果 与空白对照组相比,激动剂及缺氧复氧处理方法均诱导原代皮质神经元TLR3、TLR3mRNA表达增强(P<0.05);激动剂预处理后,与OGD组相比,激动剂+OGD组神经元细胞状态较好,数目较多(P<0.05)。结论 TLR3参与缺糖缺氧诱导的原代皮质神经元的损伤过程,激动剂可减轻神经元缺糖缺氧后损伤。

日本鳗鲡TLR3基因的克隆及其免疫功能分析

Toll样受体3(TLR3)是用于识别双链RNA的一种重要模式识别受体。利用RT-PCR和RACE技术克隆了日本鳗鲡TLR3(Aj TLR3)基因c DNA全长序列,并采用实时荧光定量PCR(q RT-PCR)检测分析该基因在日本鳗鲡各组织器官以及体外肝脏细胞的表达水平变化,以期对日本鳗鲡TLR3基因的序列特征及其功能进行研究。结果表明:Aj TLR3基因全长3 383 bp,开放阅读框为2 766 bp,编码921个氨基酸。该蛋白具有16个LRR的胞外结构域、跨膜结构域以及TIR结构域,其中在TIR结构域含有一个高度保守的氨基酸残基Tyr^(778)。Aj TLR3基因在日本鳗鲡各组织器官中均有表达,其中肝脏表达量最高;经poly I∶C刺激后在血、肠、肝脏、脾脏、皮肤、心脏及肌肉组织中均显著提高;而LPS刺激后,仅在肝脏和肠中有显著提高。日本鳗鲡肝脏细胞体外实验结果显示:poly I∶C处理后12 h和24 h表达水平显著增高,Cp G-DNA和肽聚糖处理后24 h表达量均有显著增加;而细菌浓度达到107 CFU/m L和108 CFU/m L后,Aj TLR3基因表达水平分别在24 h、12 h显著增高并达到峰值。以上研究表明,Aj TLR3在日本鳗鲡抵御病毒及细菌的免疫应答过程中具有重要作用。

TLR3与年龄相关性黄斑变性疾病的相关性

年龄相关性黄斑变性(age-related macular degeneration,ARMD)是一种多发于50岁以上人群的慢性进展性疾病,是发达国家老年人群的主要致盲性眼病,也是发展中国家老年人群不可逆视力损害的主要原因,ARMD包括地图状萎缩及脉络膜新生血管两类。由于其具体病因和发病机制尚不明确,大多数学者认为这是一种多因素疾病。近年来Toll样受体3(Toll-like receptor 3,TLR3)与ARMD关系的研究成为热点,本文就近年来相关研究进展做一综述。

微小RNA-186-5p(miR-186-5p)下调TLR3表达抑制高糖诱导的心肌细胞凋亡

目的研究高糖处理AC16心肌细胞微小RNA-186-5p(miR-186-5p)、Toll样受体3(TLR3)表达水平的变化及其与心肌细胞凋亡的关系。方法靶基因预测数据库Target Scan7.1预测miR-186-5p可直接作用于TLR3,采用高糖诱导培养的人AC16心肌细胞,建立糖尿病心肌病模型。采用实时定量PCR检测心肌细胞miR-186-5p水平,Western blot法检测心肌细胞TLR3表达水平。采用细胞转染过表达或降低miR-186-5p或TLR3水平,通过流式细胞术检测心肌细胞凋亡情况,Western blot法检测心肌细胞裂解型胱天蛋白酶3(c-caspase-3)的蛋白表达,荧光素酶报告基因检测miR-186-5p与TLR3的相互作用。结果生物信息学分析和荧光素酶活性测定证实TLR3为miR-186-5p直接作用的靶基因,高糖处理的心肌细胞miR-186-5p表达下调,过表达miR-186-5p可逆转高糖诱导的细胞凋亡,并降低c-caspase-3蛋白水平。下调TLR3水平抑制高糖诱导的细胞凋亡并降低c-caspase-3蛋白水平。过表达TLR3可以上调miR-186-5p抑制高糖诱导的心肌细胞凋亡水平。TLR3表达水平升高可逆转miR-186-5p抑制HG诱导心肌细胞凋亡的作用。结论miR-186-5p通过下调TLR3表达抑制高糖诱导的心肌细胞凋亡。

TLR3对细胞凋亡相关信号通路的调控

Toll样受体3(TLR3)作为特异性模式识别受体(PRR)能特异性识别模式相关分子双链RNA(dsRNA)从而发挥免疫作用。最近研究表明TLR3还可以诱导多种细胞发生凋亡,特别是在肿瘤中TLR3信号通路更倾向诱导凋亡,这为进一步研究TLR3激动剂干预肿瘤提供了依据。

帕金森患者外周血Toll样受体3表达水平与周围神经病变的相关性研究

目的研究帕金森患者外周血Toll样受体3(Toll-like receptor 3,TLR3)表达与周围神经病变的相关性。方法选取2023年1月-12月在黑河市第一人民医院神经内科就诊并治疗的112例帕金森患者为帕金森组,纳入同期在本院进行健康体检的120例老年体检者为健康对照组。采用实时荧光定量聚合酶链反应检测所有研究对象外周血TLR3的表达水平。对帕金森患者进行肌电图检查,判断是否存在周围神经病变。采用修订的Hoehn-Yahr(H-Y)分级评估帕金森患者的病情严重程度。结果与健康对照组比较,帕金森组外周血TLR3表达水平显著升高,差异有统计学意义(t=7.341,P<0.001)。112例帕金森患者中,发生周围神经病变50例,未发生周围神经病变62例。与无周围神经病变组比较,周围神经病变组外周血TLR3表达水平显著升高,差异有统计学意义(t=4.632,P<0.001)。二元多因素Logistic回归分析结果表明,较高的H-Y分级、左旋多巴日剂量、同型半胱氨酸和外周血TLR3水平及较低的维生素B 12水平是帕金森合并周围神经病变的独立危险因素(P<0.05)。帕金森患者外周血TLR3表达水平与感觉神经神经传导速度(Nerve conductive velocity,NCV)、运动神经NCV呈负相关关系(r分别为-0.398和-0.442,P均<0.001)。ROC曲线分析表明,外周血TLR3表达水平评估帕金森病合并周围神经病变的曲线下面积为0.879(0.817~0.942),灵敏度为92.0%,特异度为71.0%。结论外周血TLR3表达水平在帕金森合并周围神经病变组中显著升高,与神经NCV显著负相关,对帕金森合并周围神经病变的诊断具有一定价值。

TLR7/8 signalling affects X-sperm motility via the GSK3α/β-hexokinase pathway for the efficient production of sexed dairy goat embryos

Background:Goat milk is very similar to human milk in terms of its abundant nutrients and ease of digestion.To derive greater economic benefit,farmers require more female offspring(does);however,the buck-to-doe offspring sex ratio is approximately 50%.At present,artificial insemination after the separation of X/Y sperm using flow cytometry is the primary means of controlling the sex of livestock offspring.However,flow cytometry has not been successfully utilised for the separation of X/Y sperm aimed at sexing control in dairy goats.Results:In this study,a novel,simple goat sperm sexing technology that activates the toll-like receptor 7/8(TLR7/8),thereby inhibiting X-sperm motility,was investigated.Our results showed that the TLR7/8 coding goat Xchromosome was expressed in approximately 50%of round spermatids in the testis and sperm,as measured from cross-sections of the epididymis and ejaculate,respectively.Importantly,TLR7/8 was located at the tail of the Xsperm.Upon TLR7/8 activation,phosphorylated forms of glycogen synthase kinaseα/β(GSK3α/β)and nuclear factor kappa-B(NF-κB)were detected in the X-sperm,causing reduced mitochondrial activity,ATP levels,and sperm motility.High-motility Y-sperm segregated to the upper layer and the low-motility X-sperm,to the lower layer.Following in vitro fertilisation using the TLR7/8-activated sperm from the lower layer,80.52±6.75%of the embryos were XX females.The TLR7/8-activated sperm were subsequently used for in vivo embryo production via the superovulatory response;nine embryos were collected from the uterus of two does that conceived.Eight of these were XX embryos,and one was an XY embryo.Conclusions:Our study reveals a novel TLR7/8 signalling mechanism that affects X-sperm motility via the GSK3α/β-hexokinase pathway;this technique could be used to facilitate the efficient production of sexed dairy goat embryos.

基于TLR3/NF-κB信号通路探讨RSV诱导COPD急性加重的分子机制

目的基于Toll样受体(TLR)3/核转录因子(NF)-κB信号通路探讨呼吸道合胞病毒(RSV)诱导慢性阻塞性肺疾病(COPD)急性加重的分子机制。方法将SD大鼠分为3组,每组6只。正常组和COPD组用烟熏法建立COPD大鼠模型后,用10%水合氯醛按0.3 ml/100 g体质量经腹腔注射麻醉,每一个鼻孔内滴入0.4μl/g的无病毒培养基;RSV+COPD组用10%水合氯醛按0.3 ml/100 g体质量经腹腔注射麻醉,在每一个鼻孔内滴入0.4μl/g滴度为5×104TCID50/0.1 ml的RSV悬液。以上操作每周重复1次。第8周取肺组织苏木素-伊红(HE)染色观察病理改变;免疫组织化染色检测TLR3在肺组织中的表达,Western印迹检测TLR3、NF-κB p65在肺组织中的蛋白表达,酶联免疫吸附试验(ELISA)检测白细胞介素(IL)-6和IL-32蛋白水平。结果RSV+COPD组体质量增长比COPD组明显减慢(P<0.05),肺部病理学切片提示炎症明显加重。RSV+COPD组肺组织TLR3的免疫阳性信号较COPD组显著增高,RSV+COPD组肺组织中TLR3、NF-κB p65、IL-6及IL-32蛋白表达较COPD组显著增高(P<0.05)。结论RSV诱发COPD急性加重的机制可能是通过上调TLR3并激活NF-κB信号通路。

TLR与PI3K-Akt通路交互调控的研究进展

Toll样受体(Toll Like Receptor,TLR)作为模式识别受体,对机体的免疫防御和稳态维持至关重要。为了保证TLR通路介导的免疫反应适时适度地进行,机体存在对TLR通路的精细的正负调控机制。磷脂酰肌醇-3-激酶(Phosphatidylinositol 3-Kinase,PI3K)-蛋白激酶B(Akt)信号通路在细胞的增殖、分化、迁移和代谢等众多生理过程中都发挥重要调控作用,也参与了TLR通路的负调控。TLR通路和PI3K-Akt通路之间存在复杂的交互调控,但细节部分一直未被充分阐明。本文对这一领域的相关文献进行综述,以期增进对TLR通路调控机制的理解。

Chaihu Longgu Muli Decoction relieving temporal lobe epilepsy in rats by inhibiting TLR4 signaling pathway through miR-146a-3p and miR-146a-5p

Objective To explore the effect and mechanism of Chaihu Longgu Muli Decoction(柴胡龙骨牡蛎汤,CHLGMLD)in rats with temporal lobe epilepsy(TLE).Methods A total of 80 Sprague-Dawley(SD)male rats were randomized into control(CON),model(MOD),carbamazepine(CBZ,0.1 g/kg),CHLGMLD low dose(CHLGMLD-L,12.5 g/kg),and high dose(CHLGMLD-H,25 g/kg)groups,with 16 rats in each group.TLE rat models were established in the four groups with the use of lithium-pilocarpine except for the CON group.After the successful establishment of TLE models,all drugs were administered through gavage,and distilled water was given to rats in the CON and MOD groups for four weeks.The frequency and duration of seizures before and after treatment were recorded for the evaluation of the alleviation degree.Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression levels of miR-146a-3p and miR-146a-5p.The expression levels of toll-like receptor 4(TLR4),interleukin-1 receptor-associated kinase 1(IRAK1),tumor necrosis factor(TNF)receptor-associated factor 6(TRAF6),TAK1-binding protein(TAB),nuclear factor-kappa B(NF-κB),and interleukin-1 beta(IL-1β)in hippocampus were tested by immunofluorescence assay.Correlation analysis between the above factors and expressions of miR-146a-3p and miR-146a-5p were performed separately.Results CHLGMLD decreased the frequency(P<0.05)and duration(P<0.01)of seizures in rats.CHLGMLD down-regulated the expression levels of miR-146a-5p and miR-146a-3p(P<0.05),and inhibited the expression levels of TLR4,IRAK1,TRAF6,TAB,NF-κB,and IL-1β(P<0.01).The correlation analysis revealed that the expression levels of TLR4,IRAK1,TRAF6,TAB,NF-κB,and IL-1β were positively correlated with the expression levels of miR-146a-3p and miR-146a-5p detected by qRT-PCR,respectively(P<0.01).Conclusion CHLGMLD can inhibite the TLR4 signaling pathway by lowering the expression levels of miR-146a-3p and miR-146a-5p to alleviate hippocampal dentate gyrus inflammation in TLE rats,thus relieving seizures.

Efficacy of a Novel Antibody TLR3 Modulator in the Self-Treatment of Common Cold: The ESTUAR Trial

Context: Since the discovery of toll-like receptor 3 (TLR3), no specific tools have been developed to modulate its activity in upper respiratory tract viral infections (URTIs). ContafluTM (antibodies to TLR3 cytoplasmic fragment) is the first specific TLR3 modulator that showed efficacy in a mouse model of influenza. Objective: To evaluate the efficacy of Contaflu in URTI. Methods: A double-blind randomized placebo-controlled trial in adults with self-reported URTI (the ESTUAR trial) was conducted in 2012/2013 in Belgium. Adult outpatients started a 7-day treatment course with oral tablets of Contaflu or placebo within 36 h after onset of at least one of 4 typical symptoms of URTI. Patients were examined twice by their general practitioners, on days 2-3 and 10-14 after start of treatment. The primary endpoint was the overall severity of URTI calculated as the sum of Wisconsin Upper Respiratory Symptom Survey (WURSS-21) scores over the follow-up. Independent Student’s t test was used to compare the disease severity between groups. Results: A total of 243 patients were enrolled by 32 investigators (121 Contaflu, 122 placebo);92% of cases matched ICD codes J00 or J06. Most patients had very mild (41.8%) or mild (18.2%) URTI symptoms. In the ITT cohort, neither primary nor secondary outcome measures (duration of URTI, day-to-day and overall functional impairments) showed statistically significant differences between groups. The rate of adverse events was similar in both groups. In patients with moderate to severe URTI symptoms, Contaflu tended to reduce the overall disease severity, daily symptoms, and to improve the functional state. Due to the small size of the corresponding subgroups, Contaflu efficacy on daily scores was statistically significant (p < 0.05) only 1, 2, and 5 days after start of treatment. Conclusion: Contaflu was ineffective in mild URTI and showed efficacy in moderate to severe URTI cases.

Effects of Combination of 1,25(OH)_(2)D_(3) and TLR-4 Inhibitor on the Damage to HaCaT Cells Caused by UVB Irradiation

Objective Vitamin D and Toll-like receptor-4(TLR-4)inhibition are involved in the protection of keratinocytes.The effects of combination of 1,25(OH)_(2)D_(3) and TLR-4 inhibitor on the protection of keratinocytes against ultraviolet radiation B(UVB)irradiation remain unclear.This study was undertaken to explore the effects of combination of 1,25(OH)_(2)D_(3) and TAK-242(TLR-4 inhibitor)on the damage to HaCaT cells caused by UVB irradiation.Methods In vitro,HaCaT cells were treated with 1,25(OH)_(2)D_(3) or/and TAK-242 prior to UVB irradiation at the intensity of 20 mJ/cm^(2),then the production of reactive oxygen species(ROS),cell migration,apoptosis of cells,and the expression of oxidative stress,endoplasmic reticulum stress,and apoptosis related proteins were determined.Results Compared with the HaCaT cells treated with 1,25(OH)_(2)D_(3) or TAK-242,the cells treated with both 1,25(OH)_(2)D_(3) and TAK-242 showed,1)significantly lower production of ROS(P<0.05);2)significantly less apoptosis of HaCaT cells(P<0.05);3)significantly lower expression of NF-κB,Caspase-8,Cyto-C,Caspase-3(P<0.05).Conclusion The combination of 1,25(OH)_(2)D_(3) and TAK-242 could produce a better protection for HaCaT cells via inhibiting the oxidative stress,endoplasmic reticulum stress and apoptosis than 1,25(OH)_(2)D_(3) or TAK-242 alone.