The Role of Toll-Like Receptors and Nuclear Factor κB p65 Protein in the Pathogenesis of Otitis Media

The role of Toll-like receptor 4 (TLR4) and nuclear factor κB p65 (NF-κB p65) proteins in the pathogenesis of otitis media is explored. In recent years, the incidence of otitis media has been rising globally, becoming a significant threat to human health. More and more studies have found that Toll-like receptor 4 (TLR4), as a member of the Toll-like receptor family, can promote the generation of inflammatory factors and is closely related to the body’s immune response and inflammatory response. Nuclear factor-κB p65 (NF-κB p65) is a nuclear transcription factor that can interact with various cytokines, growth factors, and apoptotic factors, participating in processes such as oxidative stress, apoptosis, and inflammation in the body [1]. This article elaborates on the structure, function, and signaling pathways of TLR4 and NF-κB p65 proteins in the pathogenesis of otitis media, aiming to provide more precise targets and better therapeutic efficacy for the diagnosis and treatment of otitis media. The role of inflammation in disease.

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.

左归降糖舒心方调控TLR4/NF-κB通路抑制糖尿病心肌病小鼠心肌纤维化的机制研究

目的 基于Toll样受体4(Toll-like receptor 4,TLR4)/核因子κB(nuclear factor-κB,NF-κB)信号通路探讨左归降糖舒心方对糖尿病心肌病MKR小鼠心肌纤维化及炎症因子的影响。方法 以8周龄雄性MKR小鼠为实验对象,采用高脂饮食联合腹腔注射链脲佐菌素(streptozotocin,STZ)40 mg/kg构建糖尿病心肌病模型,随机分为中药高剂量组[33.67 g/(kg·d)左归降糖舒心方2 g/mL]、中药低剂量组[16.84 g/(kg·d)左归降糖舒心方2 g/mL]、西药联合组[0.23 g/(kg·d)二甲双胍联合1.5 mg/(kg·d)依那普利]和模型组(等容量蒸馏水);另设FVB小鼠为空白对照组(等容量蒸馏水)。每组10只,连续给药8周后取材。收集尾静脉血液检测空腹血糖水平;采用HE、Masson染色观察心肌病理改变,电镜观察心肌组织超微结构变化;采用ELISA检测小鼠血清肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)及白细胞介素-1β(interleukin-1β,IL-1β)含量;采用Western blot法检测TLR4、NF-κB p56、p-NF-κB p56/NF-κB p56蛋白表达情况;RT-qPCR及Western blot检测小鼠心肌组织Ⅰ型胶原(collagen type I,CollagenⅠ)、Ⅲ型胶原(collagen typeⅢ,CollagenⅢ)及α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)的表达水平。结果 与空白对照组比较,模型组小鼠空腹血糖及血清中TNF-α、IL-1β含量增高(P<0.01);心肌细胞结构紊乱,胶原含量增加,心肌细胞重度退行性变;心肌组织中TLR4、NF-κB p56、p-NF-κB p56/NF-κB p56蛋白表达上调(P<0.01),CollagenⅠ、CollagenⅢ、α-SMA mRNA及蛋白表达增加(P<0.05,P<0.01),中药高剂量组小鼠心肌组织CollagenⅠ、CollagenⅢ蛋白表达增加(P<0.01)。与模型组比较,中药高、低剂量组和西药联合组小鼠空腹血糖及血清中TNF-α、IL-1β含量均降低(P<0.01);心肌结构改善,胶原沉积减少;心肌组织中TLR4、NF-κB p56、p-NF-κB p56/NF-κB p56蛋白表达下调(P<0.01),CollagenⅠ、CollagenⅢ、α-SMA蛋白及mRNA表达下调(P<0.01)。与西药联合组比较,中药低剂量组小鼠空腹血糖及血清中TNF-α、IL-1β含量均升高(P<0.01),心肌结构无明显改善,胶原沉积无显著减少;心肌组织中TLR4、NF-κB p56、p-NF-κB p56/NF-κB p56蛋白表达上调(P<0.01),CollagenⅠ、CollagenⅢ、α-SMA蛋白及mRNA表达上调(P<0.01)。结论 左归降糖舒心方能抑制心肌炎性反应、改善心肌细胞结构,其作用机制可能与调控TLR4/NF-κB通路、下调细胞炎症因子表达、抑制心肌纤维化有关。

HMGB1基因敲除通过抑制TLR4/NF-κB通路减轻脓毒症小鼠急性肺损伤

目的 研究高迁移率族蛋白B1(HMGB1)基因敲除减轻脓毒症小鼠急性肺损伤及抑制Toll样受体4(TLR4)/核因子-κB(NF-κB)通路的作用。方法 野生型(WT)小鼠分为WT-Sham组和WT-模型组,HMGB1基因敲除(KO)小鼠分为KO-Sham组和KO-模型组。WT-模型组和KO-模型组采用盲肠结扎穿孔术制备脓毒症ALI模型,WT-Sham组和KO-Sham组进行假手术操作。造模后24 h,检测动脉血氧分压(PaO_(2)),计算氧合指数(OI),检测肺组织病理改变,计算肺损伤评分,检测血清及肺组织中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、IL-6、活性氧簇(ROS)、丙二醛(MDA)、超氧化物歧化酶(SOD)的浓度,肺组织中HMGB1、TLR4、核NF-κB的表达。结果 WT-模型组的PaO_(2)、OI、血清及肺组织SOD的浓度低于WT-Sham组,肺损伤评分、血清及肺组织中TNF-α、IL-1β、IL-6、ROS、MDA的浓度、肺组织中HMGB1、TLR4、核NF-κB的表达水平高于WT-Sham组(P<0.05);KO-模型组肺组织中不表达HMGB1,PaO_(2)、OI、血清及肺组织SOD的浓度高于WT-模型组,肺损伤评分、血清及肺组织中TNF-α、IL-1β、IL-6、ROS、MDA的浓度、肺组织中TLR4、核NF-κB的表达水平低于WT-模型组(P<0.05)。结论敲除HMGB1减轻脓毒症小鼠ALI,相关的分子机制可能是抑制TLR4/NF-κB通路介导的炎症反应和氧化应激反应。

五味子乙素通过TLR4/NF-κB信号通路对急性胰腺炎大鼠肺部损伤的影响

目的:探讨五味子乙素通过Toll样受体4(TLR4)/核转录因子-κB(NF-κB)信号通路对急性胰腺炎(AP)大鼠肺部损伤的影响。方法:取SD大鼠,通过胆胰管内逆行注射5%牛磺胆酸钠方法诱导建立AP肺损伤模型,经随机数表法分为模型组、五味子乙素组、TLR4过表达载体组、TLR4空载组、五味子乙素+TLR4过表达载体组,每组12只大鼠,再取12只SD大鼠仅翻动肠管不注射5%牛磺胆酸钠,作为假手术组。以药物分别干预大鼠后,检测各组大鼠肺功能及各组大鼠腹水量与肺组织湿重/干重(W/D);HE染色检测各组大鼠肺组织病理形态并评分;检测各组大鼠动脉血气;全自动生化分析仪检测大鼠血清淀粉酶,ELISA检测炎症细胞因子IL-6、IL-18水平;蛋白免疫印迹法检测肺组织TLR4/NF-κB通路蛋白表达;免疫组织化学染色检测肺组织TLR4蛋白表达。结果:与假手术组相比,模型组大鼠肺组织出现病理损伤改变,模型组大鼠MV、PEF、PaO_(2)、OI显著降低(P<0.05),Ri、腹水量与W/D、PaCO_(2)、Holfbauer评分、血清淀粉酶、IL-6与IL-18水平、肺组织TLR4阳性细胞比例、TLR4与MYD88蛋白表达、p-NF-κB p65/NF-κB p65水平显著升高(P<0.05)。与模型组、五味子乙素+TLR4过表达载体组分别相比,五味子乙素组大鼠肺组织病理损伤改变程度均减轻,MV、PEF、PaO_(2)、OI均升高(P<0.05),Ri、腹水量与W/D、PaCO_(2)、Holfbauer评分、血清淀粉酶、IL-6与IL-18水平、肺组织TLR4阳性细胞比例、TLR4与MYD88蛋白表达、p-NF-κB p65/NF-κB p65水平均降低(P<0.05);TLR4过表达载体组大鼠肺组织病理损伤改变程度均加重,MV、PEF、PaO_(2)、OI均降低(P<0.05),Ri、腹水量与W/D、PaCO_(2)、Holfbauer评分、血清淀粉酶、IL-6与IL-18水平、肺组织TLR4阳性细胞比例、TLR4与MYD88蛋白表达、p-NF-κB p65/NF-κB p65水平均升高(P<0.05)。与模型组相比,TLR4空载组大鼠各指标差异无统计学意义(P>0.05)。结论:五味子乙素可通过下调TLR4/NF-κB信号通路,抑制炎症,减轻AP大鼠肺部损伤,修复肺功能。

基于TLR4/NF-κB通路调补肺肾法干预细胞自噬对COPD肺血管重塑的影响和机制

目的基于Toll样受体4(TLR4)/核转录因子-κB(NF-κB)通路研究调补肺肾法干预细胞自噬对慢性阻塞性肺疾病(COPD)肺血管重塑的影响和机制。方法取SD大鼠采用香烟烟雾暴露结合反复细菌感染的方案建立COPD模型,随机分为3组,模型组、补肺益肾方(3.7 g/kg)组、补肺益肾方(3.7 g/kg)+脂多糖(LPS)(TLR4激活剂,15 mg/kg)组,每组12只,另取12只大鼠正常呼吸并气管滴注等剂量生理盐水作为对照组,经补肺益肾方、LPS对大鼠分组干预后,检测各组大鼠肺功能指标:潮气量(TV)、呼气峰流速(PEF)、第0.3秒用力呼气容积(FEV0.3)/用力肺活量(FVC)。以HE染色检测各组大鼠肺组织病理形态和肺血管重塑,比较其管壁厚度(WT)占血管直径(VD)百分比WT/VD(%)、管腔面积(LA)占血管总面积(TA)百分比LA/TA(%)。以免疫荧光染色检测各组大鼠肺组织内肺血管内皮标记物CD34表达。以酶标仪检测各组大鼠肺泡灌洗液(BALF)及血清促炎因子:肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-17水平。以免疫印迹法检测各组大鼠肺组织自噬及TLR4/NF-κB信号通路相关蛋白表达。结果与对照组相比,模型组大鼠肺组织呈现明显病理损伤及肺血管重塑症状,TV、PEF、FEV0.3/FVC、LA/TA、肺组织Beclin-1蛋白表达与LC3Ⅱ/LC3Ⅰ显著降低(P<0.05),WT/VD、CD34相对阳性表达、BALF及血清促炎因子TNF-α与IL-17水平、肺组织TLR4蛋白表达与p-NF-κB p65/NF-κB p65显著升高(P<0.05)。与模型组相比,补肺益肾方组大鼠肺组织损伤及肺血管重塑症状减轻,TV、PEF、FEV0.3/FVC、LA/TA、肺组织Beclin-1蛋白表达与LC3Ⅱ/LC3Ⅰ升高(P<0.05),WT/VD、CD34相对阳性表达、BALF及血清促炎因子TNF-α与IL-17水平、肺组织TLR4蛋白表达与p-NF-κB p65/NF-κB p65降低(P<0.05)。与补肺益肾方组相比,补肺益肾方+LPS组大鼠肺组织损伤及肺血管重塑症状加重,TV、PEF、FEV0.3/FVC、LA/TA、肺组织Beclin-1蛋白表达与LC3Ⅱ/LC3Ⅰ降低(P<0.05),WT/VD、CD34相对阳性表达、BALF及血清促炎因子TNF-α与IL-17水平、肺组织TLR4蛋白表达与p-NF-κB p65/NF-κB p65升高(P<0.05)。结论补肺益肾方可通过抑制TLR4/NF-κB信号而抑制COPD体内炎症,并增强自噬,进而减轻大鼠肺组织病理损伤和肺血管重塑,改善其肺功能。

Efficacy of Jiangzhi Xiaoban tablet(降脂消斑片)on toll-like receptor 4/nuclear factor-kappa B/nod-like receptor protein 3 signaling pathway in mice with atherosclerosis induced by high-fat diet

OBJECTIVE:To study the effect of Jiangzhi Xiaoban tablet(降脂消斑片,JZXB)on toll-like receptor 4(TLR4)/nuclear factor-kappa B(NF-κB)/Nod-like receptor protein 3(NLRP3)signaling pathway expression in atherosclerosis(AS)mice by establishing a mouse model of AS,and to explore its mechanism of prevention and treatment of AS.METHODS:Sixty-four male C57BL/6J mice were randomly divided into two groups,12 in the normal control group and 52 in the model group(MOD).Seven weeks later,two mice in each of the above two groups were randomly sacrificed,and the whole aortic tissue of the mice was taken out for hematoxylin-eosin staining.After successful modeling,50 mice in the modeling group were randomly divided into 5 groups:MOD,atorvastatin group(ATO),low-dose group of JZXB(JZXB-L),middle-dose group of JZXB(JZXB-M),and high-dose group of JZXB(JZXB-H),10 mice in each group.The mice in each group were killed after 6 weeks of preventive administration.HE staining was used to observe the pathological changes of aorta in AS mice.The levels of serum triglyceride(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C)and high-density lipoprotein cholesterol(HDL-C)were detected by automatic biochemical analyzer.The levels of inflammatory factor interleukin-1β(IL-1β)were detected by enzyme linked immunosorbent assay.The expression of TLR4,NF-κB and NLRP3 proteins in aortic tissue was detected by immunohistochemistry.RESULTS:Compared with the MOD,the levels of serum TC,TG and LDL-C in the JZXB-H and ATO were significantly decreased,while the level of HDL-C was significantly increased.The levels of serum TG,LDL-C in the JZXB-M were significantly decreased,and the level of HDL-C was significantly increased.Compared with the MOD,the levels of IL-1βwere significantly decreased,aortic lesions were significantly improved,and the expression of TLR4,NF-κB,and NLRP3 proteins in the aortic tissue was significantly decreased in the JZXB-H,JZXB-M,and ATO.CONCLUSION:JZXB has inhibitory effect on atherosclerosis in mice,and its mechanism may be through regulating the TLR4/NF-κB/NLRP3 signaling pathway and reducing the inflammatory response,so as to play a role in inhibiting atherosclerosis.

Protective effect of modified Huangqi Chifeng decoction(加味黄芪赤风汤)on immunoglobulin A nephropathy through toll-like receptor 4/myeloid differentiation factor 88/nuclear factor-kappa B signaling pathway

OBJECTIVE:To examine the nephroprotective mechanism of modified Huangqi Chifeng decoction(加味黄芪赤风汤,MHCD)in immunoglobulin A nephropathy(IgAN)rats.METHODS:To establish the IgAN rat model,the bovine serum albumin,lipopolysaccharide,and carbon tetrachloride 4 method was employed.The rats were then randomly assigned to the control,model,telmisartan,and high-,medium-,and low-dose MHCD groups,and were administered the respective treatments via intragastric administration for 8 weeks.The levels of 24-h urinary protein,serum creatinine(CRE),and blood urea nitrogen(BUN)were measured in each group.Pathological alterations were detected.IgA deposition was visualized through the use of immunofluorescence staining.The ultrastructure of the kidney was observed using a transmission electron microscope.The expression levels of interleukin-6(IL-6),monocyte chemoattractant protein-1(MCP-1),and transforming growth factor-β1(TGF-β1)were examined by immunohistochemistry and quantitative polymerase chain reaction.Levels of toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),and nuclear factor-kappa B(NF-κB)P65,were examined by immunohistochemistry,Western blotting,and quantitative polymerase chain reaction.RESULTS:The 24-h urine protein level in each group increased significantly at week 6,and worsen from then on.But this process can be reversed by treatments of telmisartan,and high-,medium-,and low-dose of MHCD,and these treatments did not affect renal function.Telmisartan,and high-,and medium-dose of MHCD reduced IgA deposition.Renal histopathology demonstrated the protective effect of high-,medium-,and low-dose of MHCD against kidney injury.The expression levels of MCP-1,IL-6,and TGF-β1 in kidney tissues were downregulated by low,medium and high doses of MHCD treatment.Additionally,treatment of low,medium and high doses of MHCD decreased the protein and mRNA levels of TLR4,MyD88,and NF-κB.CONCLUSIONS:MHCD exerted nephroprotective effects on IgAN rats,and MHCD regulated the expressions of key targets in TLR4/MyD88/NF-κB signaling pathway,thereby alleviating renal inflammation by inhibiting MCP-1,IL-6 expressions,and ameliorating renal fibrosis by inhibiting TGF-β1 expression.

血清转化生长因子β1、白细胞介素-6、Toll样受体-4、核转录因子κB联合评估非小细胞肺癌放射性肺炎病情严重程度的价值

目的探讨血清转化生长因子β1(TGF-β1)、白细胞介素-6(IL-6)、Toll样受体-4(TLR-4)和核转录因子κB(NF-κB)联合评估非小细胞肺癌(NSCLC)放射性肺炎(RP)病情严重程度的价值。方法选取104例NSCLC放疗后继发RP患者作为研究组,另选取52例NSCLC放疗后未继发RP患者作为对照组。比较2组患者放疗前后血清TGF-β1、IL-6、TLR-4、NF-κB水平。比较研究组不同程度RP患者放疗前后血清TGF-β1、IL-6、TLR-4、NF-κB水平,分析各血清指标单独及联合评估NSCLC放疗后RP病情程度的价值。结果放疗结束后,研究组患者血清TGF-β1、IL-6、TLR-4、NF-κB水平均高于对照组,差异有统计学意义(P<0.05);放疗结束后,随着RP病情程度的增加,研究组患者血清TGF-β1、IL-6、TLR-4、NF-κB水平呈升高趋势,差异有统计学意义(P<0.05)。受试者工作特征(ROC)曲线分析结果显示,放疗结束后血清TGF-β1、IL-6、TLR-4、NF-κB水平评估1级RP的曲线下面积(AUC)分别为0.787、0.718、0.783、0.801,评估≥2级RP的AUC分别为0.729、0.740、0.793、0.825;血清TGF-β1、IL-6、TLR-4、NF-κB阳性表达患者发生≥2级RP的风险分别是阴性表达患者的2.473、2.275、2.610、5.267倍(P<0.05);血清TGF-β1、IL-6、TLR-4、NF-κB联合评估≥2级RP的AUC为0.939,敏感度为76.00%,特异度为97.47%。结论NSCLC患者放疗结束后血清TGF-β1、IL-6、TLR-4、NF-κB水平均与RP病情严重程度呈正相关,四者联合评估≥2级RP的价值显著,可为临床控制RP病情提供参考依据。

温阳化痰贴对哮喘患儿调控因子TLR4/NF-κB影响研究

目的研究温阳化痰贴对哮喘患儿调控因子Toll样受体4(TLR4)/核转录因子-κB(NF-κB)的影响。方法选取2020年10月至2021年6月北京市昌平区中西医结合医院儿科门诊治疗的小儿哮喘缓解期患儿100例,随机分为对照组51例、治疗组49例,对照组给予西医常规治疗,治疗组西医治疗联合应用温阳化痰贴贴敷。结果治疗组的调控因子TLR4、NF-κB及肺功能FEF25、FEF50、FEF75较对照组明显改善,差异有统计学意义(P<0.05)。结论应用温阳化痰穴贴联合西药治疗小儿哮喘可以调节机体免疫调控因子,从而直接或间接调节细胞、体液免疫功能,提高机体抵抗力,使疾病快速恢复。

MAPK通过调控TLR4/Myd88/NF-κB通路对妊娠期肠梗阻胃肠功能的影响

目的 分析MAPK通过调控TLR4/MyD88/NF-κB通路对妊娠期肠梗阻胃肠功能的影响。方法 选取105只SPF级Wistar大鼠,雌性、雄性分别为70、35只,将所有大鼠同笼放置后,共有33只雌性大鼠妊娠成功,将妊娠成功的8只作为空白组,剩余25只雌性大鼠建立肠梗阻模型,最终建模成功24只,将其分为模型组、上调组、下调组各8只。结果 与模型组相比,上调组MAPK表达量、VIP、GAS、TNF-α、IL-6水平、TLR4、Myd88、NF-κB mRNA表达量及相对表达量上升,MOT、IL-4、IL-10水平下降,下调组MAPK表达量、VIP、GAS、TNF-α、IL-6水平、TLR4、Myd88、NF-κB mRNA表达量及相对表达量下降,MOT、IL-4、IL-10水平上升(P <0.05)。结论 妊娠期肠梗阻大鼠经下调MAPK干预后胃肠功能改善,炎性反应减轻,其机制可能与TLR4/Myd88/NF-κB通路得到抑制有关。

Methylprednisolone inhibits activated CD4^+ T cell survival promoted by toll-like receptor ligands

BACKGROUND: Methylprednisolone (MP) can affect the survival of CD4(+) T lymphocytes and plays an important role in adaptive immune responses; however, its mechanism of action is not clear. Recent studies have shown that toll-like receptors (TLRs) on CD4(+) T cells can directly modulate adaptive immune responses by affecting the survival and proliferation of activated CD4(+) T cells. This study aimed to investigate the relationship between MP, TLRs and activated CD4(+) T cells. METHODS: We separated and purified CD4(+) T cells from mice, activated them in vitro, and co-cultured them with TLR ligands, MP or inhibitors of nuclear factor-kappa B (NF-kappa B) and activator protein 1 (AP-1). We then assessed CD4(+) T cell survival and proliferation and the expression of NF-kappa B and AP-1. RESULTS: Activated CD4(+) T cells showed increased TLR-3 and TLR-9 mRNA expression, but polyinosinic-polycytidylic acid (poly I:C) and MP had no effect on the expression of these mRNAs. Still, poly I:C and CpG oligodeoxynucleotides (CpG DNA) increased the survival of activated CD4(+) T cells, whereas MP reduced the survival of activated CD4(+) T cells and could inhibit the survival effects of poly I:C and CpG DNA. The NF-kappa B essential modifier-binding domain (NBD) inhibited the survival of activated CD4(+) T cells induced by poly I:C and CpG DNA, but the AP-1 inhibitor crucumin did not have the same effect. The increased expression of NF-kappa B induced by poly I:C and CpG DNA in activated CD4(+) T cells could be inhibited by MP, but the same was not true for the increased expression of AP-1 induced by poly I:C and CpG DNA. Finally, the proliferation of activated CD4(+) T cells was not affected by poly I:C or MP. CONCLUSION: The survival of activated CD4(+) T cells is promoted by TLR ligands, but this effect is inhibited by MP.

Effect of remifentanil on toll-like receptor 4, NF-κB and IL-6 in rabbit myocardial ischemia/reperfusion model

Objective： To investigate whether remifentanil induced cardioprotecting effect is associated with expression of toll-like receptor 4 （TLR4）, nuclear factor rB （NF-r.B） and serum interleukin -6 （IL-6）. Methods： Fifty rabbits were randomly divided into 5 groups （n=10） according to the treatment： sham operation group （group A）, ischemla-reperfusion group （group B）, low-dose remifentanil group （group C）, mediate-dose remifentanil group （group D）, and high-dose remlfentanil group （group E） Myocardial TLR4 mRNA levels, NF-r.B protein expression and serum levels of IL-6 were observed in 120 min after reperfusion. Results： The myocardial expressions of TLR4 mRNA, NF-rd3 protein and IL-6 level in sera of groups B, C, D and E were elevated compared with group A. However, remifentanil significantly reduced the levels of TLR4 mRNA, NF- r.B protein expression and serum IL-6 in groups C, D and E compared with group B. There were remarkable differences between the groups （P〈O.O1）. Conclusion： Intravenous remifentanil has protective effect against rabbit myocardial ischemia/reperfusion injury. This effect may be associated with TLR4, NF-r.B expressions on myocytes and serum level of IL-6 in a dose-dependent manner

银杏二萜内酯葡胺在超时间窗AIS中的治疗效果及对患者血清TLR4/NF-κB信号通路因子水平的影响

目的观察银杏二萜内酯葡胺(GDLG)治疗超时间窗急性缺血性脑卒中(AIS)的效果,并探讨其对患者血清Toll样受体4(TLR4)/核转录因子-κB(NF-κB)信号通路因子水平的影响。方法选取2020年1月至2023年1月郑州大学第一附属医院收治的170例超时间窗AIS患者作为研究对象,采用电脑随机数表法分为对照组和研究组各85例。对照组患者给予常规治疗,研究组患者在常规治疗基础上给予GDLG治疗,均持续治疗两周。比较两组患者的治疗效果、治疗前、治疗1周、2周后脑血流灌注指标[脑血容量(CBV)、脑血流量(CBF)、平均通过时间(MTT)]、血清神经损伤标志物[基质金属蛋白酶抑制物-1(TIMP-1)、神经元特异性烯醇化酶(NSE)、半乳糖凝集素-3(Galectin-3)]、TLR4/NF-κB信号通路因子(TLR4、NF-κB)水平、神经功能、日常生活能力,统计比较两组治疗期间不良反应及治疗后第3个月预后情况。结果研究组患者的治疗总有效率为95.29%,明显高于对照组的84.71%,差异有统计学意义(P<0.05);治疗1周、2周后,研究组患者的CBF分别为(82.69±12.54)mL/100 g、(85.17±12.36)mL/100 g,明显高于对照组的(77.01±11.86)mL/100 g、79.24±12.10)mL/100 g,CBV分别为(6.84±1.22)mL/(100 g·min)、(7.01±1.28)mL/(100 g·min),明显高于对照组的(6.05±1.17)mL/(100 g·min)、(6.19±1.23)mL/(100 g·min),MTT分别为(9.16±1.73)s、(8.92±1.65)s,明显低于对照组的(10.02±1.85)s、(9.84±1.71)s,差异均有统计学意义(P<0.05);治疗1周、2周后,研究组患者的血清NSE水平分别为(13.75±2.81)μg/L、(12.24±2.59)μg/L,明显低于对照组的(16.29±3.68)μg/L、(15.36±3.42)μg/L,TIMP-1水平分别为(72.26±16.73)ng/mL、(68.19±14.84)ng/mL,明显低于对照组的(83.51±18.20)ng/mL、(79.81±15.72)ng/mL,Galectin-3水平分别为(4.12±1.04)ng/mL、(3.75±0.96)ng/mL,明显低于对照组的(5.03±1.08)ng/mL、(4.69±1.02)ng/mL,差异均有统计学意义(P<0.05);治疗1周、2周后,研究组患者的血清TLR4水平分别为(2.61±0.78)ng/mL、(2.39±0.74)ng/mL,明显低于对照组的(3.42±0.85)ng/mL、(3.25±0.81)ng/mL,NF-κB水平分别为(108.69±21.36)ng/mL、(98.74±18.65)ng/mL,明显低于对照组的(121.54±22.06)ng/mL、(110.23±20.67)ng/mL,差异均具有统计学意义(P<0.05);治疗1周、2周后,研究组患者的美国国立卫生院卒中量表(NIHSS)评分分别为(10.19±2.18)分、(9.21±2.04)分,明显低于对照组的(44.26±4.05)分、(62.51±4.36)分,改良Barthel指数(MBI)评分分别为(47.39±4.28)分、(65.40±4.71)分,明显高于对照组的(44.26±4.05)分、(62.51±4.36)分,差异均有统计学意义(P<0.05);治疗期间,研究组患者的不良反应总发生率为10.59%,略高于对照组的7.06%,但差异无统计学意义(P>0.05);治疗后第3个月,研究组患者的预后良好率为64.71%,明显高于对照组的48.24%,差异有统计学意义(P<0.05)。结论超时间窗AIS患者在常规治疗基础上联合GDLG治疗能明显提高治疗效果,且能更有效下调血清TLR4/NF-κB信号通路因子水平。

SGLT2抑制剂联合新活素治疗高血压伴心力衰竭的疗效及对患者TLR4/NF-κB信号通路指标的影响

目的研究钠-葡萄糖协同转运蛋白2(SGLT2)抑制剂(达格列净)联合新活素治疗高血压伴心力衰竭(心衰)的临床效果及对患者外周血单核细胞Toll样受体4/核转录因子-κB(TLR4/NF-κB)信号通路指标的影响。方法前瞻性选取2021年5月至2023年6月郑州大学第一附属医院收治的138例高血压伴心衰患者作为研究对象,采用随机数表法分为对照组、单一组和联合组各46例。对照组接受常规基础治疗,单一组在此基础上接受新活素治疗,联合组在此基础上接受达格列净联合新活素治疗。治疗7 d后比较三组患者的临床疗效,以及治疗前后的心功能[左室收缩末期内径(LVESD)、左室射血分数(LVEF)、心率(HR)]、运动耐力[6 min步行试验距离(6 MWT)]、血压[收缩压(SBP)、舒张压(DBP)]、氧化应激指标[丙二醛(MDA)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)]和TLR4/NF-κB信号通路相关指标[TLR4、NF-κB、白细胞介素-1β(IL-1β)、IL-6、肿瘤坏死因子-α(TNF-α)],同时比较三组患者的不良反应发生情况。结果联合组患者的治疗总有效率为95.65%,明显高于单一组的86.96%和对照组的73.91%,差异均有统计学意义(P<0.05);治疗后,联合组患者的LVEF、6 MWT分别为(43.28±3.16)%、(336.48±32.49)m,明显高于单一组的(40.54±2.68)%、(306.45±40.26)m和对照组的(38.07±2.89)%、(280.69±36.98)m,LVESD、HR分别为(35.29±4.26)mm、(71.34±3.62)次/min,明显低于单一组的(38.75±3.64)mm、(74.25±3.59)次/min和对照组的(42.36±4.05)mm、(76.46±3.98)次/min,且单一组变化幅度大于对照组,差异均有统计学意义(P<0.05);治疗后,联合组患者的SBP、DBP分别为(110.36±5.02)mmHg、(81.42±2.56)mmHg,明显低于单一组的(114.83±4.76)mmHg、(84.39±2.88)mmHg和对照组的(117.25±5.18)mmHg、(86.05±2.62)mmHg,且单一组低于对照组,差异均有统计学意义(P<0.05);治疗后,联合组患者的GSH-Px、CAT分别为(105.24±20.29)U/g·Hb、(148.35±32.46)U/mL,明显高于单一组的(90.35±19.45)U/g·Hb、(125.64±26.49)U/mL和对照组的(84.49±17.32)U/g·Hb、(103.29±25.38)U/mL,MDA为(7.39±2.56)mmol/L,明显低于单一组的(11.35±2.95)mmol/L和对照组的(13.08±2.72)mmol/L,且单一组变化幅度大于对照组,差异均有统计学意义(P<0.05);治疗后,联合组患者的TLR4 mRNA、NF-κB mRNA、IL-1βmRNA、IL-6 mRNA、TNF-αmRNA明显低于单一组和对照组,且单一组低于对照组,差异均有统计学意义(P<0.05);三组患者的不良反应总发生率比较差异无统计学意义(P>0.05)。结论SGLT2抑制剂联合新活素治疗高血压伴心衰能有效改善患者的氧化应激水平,调节TLR4/NF-κB信号通路,恢复患者血压与心功能,临床应用效果显著,且安全性较高。

肉苁蓉多糖通过影响肠道菌群抑制Toll样受体4/核因子-κB途径改善小鼠糖尿病肾病

旨在探究肉苁蓉多糖(Cistanche deserticola polysaccharides,CDPs)对糖尿病肾病(diabetic nephropathy,DN)小鼠模型的改善作用及其可能机制。通过高糖高脂饮食结合链霉佐菌素注射建立DN模型,将36只C57BL/6N雄性小鼠随机分组,实验组给予CDPs不同剂量处理,持续4周。检测小鼠的生理指标、肾功能、炎症因子和肠道菌群变化。结果显示,CDPs可显著改善DN小鼠的一般状态和肾脏病理结构,降低血糖、尿蛋白等指标,减少炎症因子的水平。此外,CDPs可影响肠道菌群平衡,通过增加有益菌相对丰度、降低潜在致病菌、改善肠道屏障功能。CDPs还可抑制Toll样受体4/核因子-κB信号通路的活化,减少脂多糖的释放。综上,CDPs通过影响肠道菌群和抑制炎症信号通路,对DN小鼠模型表现出改善作用,结果可为DN治疗提供新的思路。

电针对脑缺血再灌注损伤大鼠Toll样受体4/核因子κB信号通路的影响

背景:炎症反应是诱发脑缺血再灌注损伤的重要因素之一。研究表明电针可有效降低缺血性脑卒中后炎症反应,改善神经功能缺损症状,但机制尚未明确。目的:观察电针对脑缺血再灌注损伤大鼠Toll样受体4/核因子κB信号通路及炎性因子表达的影响。方法:48只雄性SD大鼠随机分为假手术组、模型组和电针组,每组16只,采用大脑中动脉线栓阻断法制备脑缺血再灌注损伤大鼠模型。造模24 h后对电针组大鼠行电针干预,每天1次,每次20 min,共5 d;假手术组和模型组不做任何干预。干预5 d后根据Longa法评定各组大鼠神经功能损伤程度;TTC染色和苏木精-伊红染色分别观察大鼠脑梗死体积及脑组织病理形态,荧光定量PCR和Western blot分别检测大脑皮质中Toll样受体4、核因子κB mRNA和蛋白表达;酶联免疫法检测血清中炎性因子白细胞介素6、白细胞介素18和肿瘤坏死因子α水平。结果与结论:①与假手术组比较,模型组大鼠神经功能评分、血清白细胞介素6、白细胞介素18、肿瘤坏死因子α水平、大脑皮质Toll样受体4、核因子κB mRNA及蛋白表达水平明显升高(P<0.01);与模型组比较,电针组大鼠神经功能评分、大脑皮质Toll样受体4、核因子κB mRNA及蛋白表达,血清白细胞介素6、白细胞介素18、肿瘤坏死因子α水平明显降低(P<0.05,P<0.01);②与假手术组比较,模型组大鼠脑梗死体积明显增加(P<0.01);与模型组比较,电针组大鼠脑梗死体积减小(P<0.05);③模型组神经元排列紊乱,部分神经细胞消失,胞核固缩,结构不完整;电针组大脑皮质神经元变性及神经细胞数量丢失程度均有不同程度减轻;④结果表明,电针能明显改善脑缺血再灌注损伤模型大鼠的神经行为学,减轻脑组织损伤,有效降低血清炎性因子水平,其机制可能与抑制Toll样受体4/核因子κB信号通路有关。

高压氧配合中药汤剂治疗中青年脑卒中患者的效果及对TLR4/NF-κB信号通路与免疫炎症因子的影响

目的 探讨高压氧配合中药汤剂治疗中青年脑卒中患者的临床疗效,并分析其对免疫炎症因子、Toll样受体4(TLR4)/核转录因子-κB(NF-κB)信号通路的影响。方法 选取2021年2月至2023年2月郑州颐和医院收治的80例中青年脑卒中患者为研究对象,遵循随机单双数分配原则分为单一组、联合组,每组40例。单一组接受中药汤剂治疗,联合组接受高压氧配合中药汤剂治疗。统计对比两组临床疗效、中医证候积分、不良反应发生情况及神经功能[美国国立卫生研究院卒中量表(NIHSS)]、日常生活能力[Barthel指数(BI)]、肢体运动功能[Fugl-Meyer运动功能量表(FMA)]。对比两组治疗前后血清免疫炎症因子[肿瘤坏死因子-α(TNF-α)、超敏C-反应蛋白(hs-CRP)、红细胞C3b受体花环率(RBC-C3bRR)、红细胞免疫复合物花环率(RBC-ICR)]、Toll样受体4(TLR4)/核转录因子-κB(NF-κB)信号通路相关因子(TLR4 mRNA、NF-κB p65 mRNA)水平。结果 联合组总有效率高于单一组(P<0.05);治疗后,联合组中医证候积分、NIHSS评分低于单一组,BI、FMA评分高于单一组(P<0.05);治疗后,联合组血清TNF-α、hs-CRP水平及RBC-ICR低于单一组,RBC-C3bRR高于单一组(P<0.05);治疗后,联合组外周血单核细胞中TLR4 mRNA、NF-κB p65 mRNA水平低于单一组(P<0.05);两组不良反应发生率比较差异无统计学意义(P>0.05)。结论 高压氧配合中药汤剂治疗中青年脑卒中患者的效果确切且具有一定安全性,可改善临床症状,减轻神经功能损伤,促进肢体功能、日常生活能力恢复,并可抑制炎症反应,改善免疫功能,可能与抑制TLR4/NF-κB信号通路活化有关。

基于HMGB1/TLR4/NF-κB信号通路研究枇杷叶提取物对急性肺损伤模型大鼠炎症因子的影响

目的 基于高迁移率族蛋白B1(HMGB1)/Toll样受体4(TLR4)/核因子-κB(NF-κB)信号通路研究枇杷叶提取物对急性肺损伤(ALI)大鼠炎症因子的影响及潜在机制。方法 60只SPF级Wistar大鼠按照随机数字表法分为空白组(NC组)、模型组(ALI组)、阳性对照组(Dex组)、枇杷叶提取物低剂量组(EJLE-L组)、枇杷叶提取物中剂量组(EJLE-M组)和枇杷叶提取物高剂量组(EJLE-H组),每组10只。ALI组、Dex组、EJLE-L组、EJLE-M组、EJLE-H组气管内均滴注2 mg/kg脂多糖建立大鼠ALI模型。建模后,Dex组腹腔注射5 mg/kg地塞米松,EJLE-L组、EJLE-M组、EJLE-H组分别腹腔注射50、100、150 mg/kg枇杷叶提取物,NC组和ALI组腹腔注射5 mg/kg生理盐水。观察各组大鼠肺组织病理组织学及细胞超微结构变化,比较各组大鼠肺组织湿重/干重值,外周循环、肺组织炎症因子白介素(IL)-1β、IL-6、肿瘤坏死因子(TNF)-α水平及肺组织HMGB1、TLR4、NF-κB累积光密度值(IOD)及蛋白、mRNA表达量。结果 与ALI组比较,Dex组、EJLE-L组、EJLE-M组、EJLE-H组肺组织湿重/干重值[(6.12±0.14)、(5.76±0.11)、(5.09±0.12)、(4.67±0.10)比(6.89±0.17),P<0.05],肺组织病理评分[(7.79±0.65)分、(5.45±0.43)分、(3.23±0.29)分、(1.02±0.20)分比(13.23±1.00)分,P<0.05],外周循环炎症因子IL-1β[(102.32±11.23)ng/L、(87.86±9.09)ng/L、(64.54±6.74)ng/L、(43.11±5.63)ng/L比(147.87±15.64)ng/L,P<0.05]、IL-6 [(114.43±15.34)ng/L、(99.07±11.23)ng/L、(78.43±9.02)ng/L、(55.46±6.12)ng/L比(156.65±19.98)ng/L,P<0.05]、TNF-α[(156.64±19.09)ng/L、(107.64±12.32)ng/L、(69.09±9.09)ng/L、(49.98±5.46)ng/L比(201.98±24.43)ng/L,P<0.05]水平,肺组织炎症因子IL-1β[(172.21±17.68)ng/L、(123.32±14.35)ng/L、(95.54±10.76)ng/L、(70.94±8.75)ng/L比(223.42±28.76)ng/L,P<0.05]、IL-6[(154.43±17.65)ng/L、(100.23±11.43)ng/L、(76.42±8.93)ng/L、(66.75±6.55)ng/L比(198.90±22.34)ng/L,P<0.05]、TNF-α[(164.58±18.97)ng/L、(110.23±14.53)ng/L、(90.23±10.98)ng/L、(61.65±6.57)ng/L比(223.42±34.52)ng/L,P<0.05]水平,肺组织HMGB1[IOD值:(232.32±34.42)、(197.87±29.09)、(123.23±15.46)、(69.98±8.98)比(398.89±45.53),蛋白:(2.32±0.29)、(2.00±0.17)、(1.78±0.10)、(1.35±0.20)比(2.76±0.21),mRNA:(2.10±0.17)、(1.85±0.19)、(1.53±0.14)、(1.29±0.20)比(2.90±0.21),P <0.05]、TLR4 [IOD值:(214.34±22.35)、(168.98±18.90)、(100.34±11.23)、(77.85±8.45)比(453.34±52.32),蛋白:(2.54±0.23)、(2.04±0.22)、(1.61±0.18)、(1.46±0.14)比(2.98±0.34),mRNA:(2.03±0.25)、(1.76±0.21)、(1.54±0.14)、(1.25±0.10)比(2.76±0.22),P<0.05]、NF-κB [IOD值:(200.97±28.87)、(159.09±16.57)、(102.32±14.35)、(67.84±7.98)比(323.43±39.98),蛋白:(1.87±0.20)、(1.34±0.19)、(1.00±0.15)、(0.88±0.09)比(2.00±0.16),mRNA:(2.67±0.20)、(2.21±0.19)、(1.97±0.17)、(1.54±0.12)比(3.02±0.22),P<0.05]均明显降低;与Dex组、EJLE-L组、EJLE-M组比较,EJLE-H组上述指标改善更明显(P<0.05)。结论枇杷叶提取物可明显抑制ALI大鼠外周循环及肺部炎症,其可能通过HMGB1/TLR4/NF-κB信号通路逆转肺损伤。

基于TLR4/MyD88/NF-κB通路介导的NLRP3炎性小体活性探讨振腹推拿改善CUMS模型大鼠海马组织炎性损伤的手法机制研究

目的 观察振腹推拿对抑郁症模型大鼠海马组织Toll样受体4(Toll-like Receptor 4,TLR4)/髓样分化因子88(myeloiddifferentiationfactor88,MyD88)/核因子-κB(nuclear factor kappa-B,NF-κB)通路介导的NLR蛋白3(NLR protein 3,NLRP3)炎性小体活性的影响,探讨振腹推拿改善抑郁症模型大鼠海马区炎性损伤的作用机制。方法 将40只大鼠随机分为4组,采用慢性不可预见性温和应激方法复制抑郁症大鼠模型。采用离子钙接头蛋白-1(ionized calcium binding adaptor molecule-1,Iba1)免疫荧光染色检测各组大鼠海马组织海马回(cornuammonis, CA)的小胶质细胞活化程度,采用酶联免疫吸附法法检测大鼠海马组织中肿瘤坏死因子(tumor necrosis factor, TNF)-α、白细胞介素(interleukin, IL)-1β、IL-6、IL-10的含量,采用蛋白免疫印迹法和实时荧光定量PCR(Real time-PCR,RT-PCR)法检测各组大鼠海马组织中TLR4、MyD88、磷酸化核因子-κB(phospho-nuclear factor kappa-B,p-NF-κB)、NLRP3、凋亡相关斑点样蛋白(Apoptosis-associated speck-like protein containing CARD,ASC)蛋白及其基因表达。结果 (1)与正常组比较,模型组大鼠海马组织CA1、CA2、CA3、CA4区的活化小胶质细胞占比均显著升高(P<0.01)。与模型组大鼠比较,振腹组、氟西汀组大鼠海马组织CA1、CA2、CA4区的活化小胶质细胞占比均显著降低(P<0.01),氟西汀组大鼠海马组织CA3区的活化小胶质细胞占比与模型组相比降低有统计学差异(P<0.05),振腹组大鼠海马组织CA3区的活化小胶质细胞占比与模型组相比仅有降低趋势。(2)与正常组比较,模型组大鼠海马组织TNF-α、IL-1β、IL-6、含半胱氨酸的天冬氨酸蛋白水解酶-1含量显著升高(P<0.01),IL-10含量显著降低(P<0.01)。与模型组大鼠比较,振腹组、氟西汀组大鼠IL-10含量均显著升高(P<0.01),TNF-α、IL-1β、IL-6、Caspase-1含量均显著降低(P<0.01)。(3)与正常组比较,模型组大鼠海马组织TLR4、MyD88、p-NF-κB、NLRP3、ASC的蛋白和mRNA含量均显著升高(P<0.01)。与模型组大鼠比较,氟西汀组大鼠海马组织TLR4、MyD88、p-NF-κB、NLRP3的蛋白含量均显著降低(P<0.01),ASC的蛋白含量下降有统计学意义(P<0.05);氟西汀组大鼠TLR4、MyD88、p-NF-κB、NLRP3、ASC的mRNA含量均显著降低(P<0.01)。与模型组大鼠比较,振腹组大鼠海马组织TLR4、ASC的蛋白含量下降有统计学意义(P<0.05),MyD88、p-NF-κB、NLRP3的蛋白含量显著降低(P<0.01);振腹组大鼠TLR4、MyD88、NF-κB、NLRP3的mRNA含量均显著降低(P<0.01),ASC的mRNA含量均降低有统计学意义(P<0.05)。结论 振腹推拿缓解抑郁模型大鼠抑郁样行为的其机制与抑制海马组织中TLR4/MyD88/NF-κB通路介导的NLRP3炎性小体活性、改善海马组织炎性损伤有关。