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.

Puerarin partly counteracts the inflammatory response after cerebral ischemia/reperfusion via activating the cholinergic anti-inflammatory pathway

Puerarin, a major isoflavonoid derived from the Chinese medical herb radix puerariae （Gegen）, has been reported to inhibit neuronal apoptosis and play an anti-inflammatory role in focal cerebral ischemia model rats. Recent findings regarding stroke pathophysiology have recognized that anti-inflammation is an important target for the treatment of ischemic stroke. The cholinergic anti-inflammatory pathway is a highly robust neural-immune mechanism for inflammation control. This study was to investigate whether activating the cholinergic anti-inflammatory pathway can be involved in the mechanism of inhibiting the inflammatory response during puerarin-induced cerebral ischemia/reperfusion in rats. Results showed that puerarin pretreatment （intravenous injection） re- duced the ischemic infarct volume, improved neurological deficit after cerebral ischemia/reperfusion and decreased the levels of interleukin-1β, interleukin-6 and tumor necrosis factor-a in brain tissue. Pretreatment with puerarin （intravenous injection） attenuated the inflammatory response in rats, which was accompanied by janus-activated kinase 2 （JAK2） and signal transducers and activators of transcription 3 （STAT3） activation and nuclear factor kappa B （NF-KB） inhibition. These observa- tions were inhibited by the alpha7 nicotinic acetylcholine receptor （a7nAchR） antagonist a-bungarotoxin （a-BGT）. In addition, puerarin pretreatment increased the expression of a7nAchR mRNA in ischemic cerebral tissue. These data demonstrate that puerarin pretreatment strongly protects the brain against cerebral ischemia/reperfusion injury and inhibits the inflammatory re- sponse. Our results also indicated that the anti-inflammatory effect of puerarin may partly be medi- ated through the activation of the cholinergic anti-inflammatory pathway.

Toll样受体2及其信号通路在大鼠面神经夹挫伤后的机制研究

目的探究面神经损伤对大鼠Toll样受体2(TLR2)/核转录因子κB(NF-κB)信号通路的影响。方法将面神经损伤大鼠随机分为三组,A组:正常对照组,B组:手术损伤后自然恢复的观察组,C组:手术后腹腔注射Pam3CSK4(TLR2激动剂)观察恢复。电镜比较3组面神经的表现;HE染色脑干组织观察神经元的凋亡,PCR观察TLR2、NF-κB p65 mRNA表达水平,免疫荧光观察TLR2、NF-κB的荧光表达。结果面瘫造模顺利,电镜下面神经损伤明显,C组大鼠髓鞘较B组更为破碎;脑区内神经元也表现为胶质细胞变性更多;PCR显示A组TLR2及NF-κB p65 mRNA表达低于B组和C组;免疫荧光显示B组表达的TLR2、NF-κB荧光强于A组。结论注射Pam3CSK4激活TLR2/NF-κB信号通路,加重炎症的表现使面神经功能障碍。

补中益气汤调控TLR4/NF-κB/AIM2信号通路改善自身免疫性甲状腺炎小鼠甲状腺炎症损伤的作用机制

目的:基于Toll样受体4(TLR4)/核转录因子-κB(NF-κB)/黑色素瘤缺乏因子2(AIM2)炎性小体信号通路探索补中益气汤改善自身免疫性甲状腺炎(AIT)小鼠炎症损伤的作用机制。方法:选用基因易感8周龄NOD.H-2h4小鼠120只,随机分为空白组、模型组、补中益气汤低、中、高剂量组(4.78、9.56、19.12 g·kg^(-1))、西药组(硒酵母片,3.033×10^(-5) g·kg^(-1))。各组AIT模型小鼠自由饮用0.05%碘化钠水溶液8周,建立AIT模型,空白组自由饮用蒸馏水。依据分组灌胃各给药组小鼠药液8周后取材,肉眼观察甲状腺组织肿胀情况,称量脾脏质量,酶联免疫吸附测定法(ELISA)检测血清炎症因子白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)含量,实时荧光定量聚合酶链式反应(Real-time PCR)检测甲状腺组织高迁移率族蛋白1(HMGB1)、TLR4、AIM2、NF-κB p65、凋亡相关斑点样蛋白(ASC)、胱天蛋白酶-1(Caspase-1)、IL-1β mRNA的表达,蛋白免疫印迹法(Western blot)检测甲状腺组织HMGB1、TLR4、AIM2、NF-κB p65、磷酸化(p)-NF-κB p65、ASC、Caspase-1、IL-1β蛋白的表达,免疫荧光染色观察小鼠甲状腺组织HMGB1、AIM2、NF-κB p65的蛋白表达情况。结果:与空白组比较,模型组小鼠甲状腺组织明显肿胀,脾脏质量明显增加,甲状腺组织HMGB1、TLR4、NF-κB p65、AIM2、ASC、Caspase-1、IL-1β表达显著升高(P<0.01);与模型组比较,补中益气汤各剂量组小鼠甲状腺组织肿胀情况好转,脾脏质量明显下降,甲状腺组织HMGB1、TLR4、AIM2、NF-κB p65、p-NF-κB p65、ASC、Caspase-1、IL-1β表达明显降低(P<0.05,P<0.01)。结论:补中益气汤可有效改善AIT的炎症损伤,调控TLR4/NF-κB/AIM2炎性小体信号通路的异常活化可能是其干预机制之一。

基于TLR/NF-κB信号通路探讨电针治疗失眠症模型大鼠的作用及机制

目的探究电针通过Toll样受体2/核转录因子kappa B(TLR/NF-κB)信号通路在治疗大鼠失眠症中的作用及机制。方法40只SD雄性大鼠中随机选取10只为对照组,其余大鼠建立失眠症模型后随机分为模型组、安定组及电针组,每组10只。安定组腹腔注射安定注射液0.92 mg·kg-1·d-1;电针组对双侧“内关”穴进行针刺,留针20 min,隔日1次;对照组与模型组给予等量的生理盐水。连续治疗15 d后,检测大鼠血清TNF-α、sTNF-RⅡ水平,脾脏单细胞中CD4^(+)Treg、CD4^(+)CD25^(+)Treg占比及TLR3、TLR4表达量,并检测脾脏组织中TLR/NF-κB通路相关蛋白及mRNA相对表达情况。结果与对照组比较,模型组和安定组TNF-α、sTNF-RⅡ水平,CD4^(+)CD25^(+)Treg占比及TLR3、TLR4表达升高,CD4^(+)Treg占比降低,MyD88、TAB2、NF-kB蛋白及TLR3、TLR4、MyD88、TAB2、NF-kB mRNA相对表达上调(P<0.05);与模型组比较,安定组和电针组TNF-α、sTNF-RⅡ水平,CD4^(+)CD25^(+)Treg占比及TLR3、TLR4表达降低,CD4^(+)Treg占比升高,MyD88、TAB2、NF-kB蛋白及TLR3、TLR4、MyD88、TAB2、NF-kB mRNA相对表达下调(P<0.05);与安定组比较,电针组TNF-α、sTNF-RⅡ水平,CD4^(+)CD25^(+)Treg占比及TLR3、TLR4表达降低,CD4^(+)Treg占比升高,MyD88、TAB2、NF-kB蛋白及TLR3、TLR4、MyD88、TAB2、NF-kB mRNA相对表达下调(P<0.05)。结论电针能够改善失眠症大鼠炎症反应,提高免疫能力,改善大鼠失眠症的机制可能与TLR/NF-κB信号通路有关。

Therapeutic Effect of Crocin on Diabetic Retinopathy in Rats Based on TLR4/My D88/NF-κB Pathway

Objective:To study the therapeutic effect of crocin on diabetic retinopathy(DR)in rats based on toll-like receptor 4(TLR4)/myeloid differentiation factor 88(My D88)/nuclear factor-κB(NF-κB)pathway.Methods:Thirty SPF SD rats were used in the experiment,which were randomly divided into DR group,control group and crocin group,with 10 rats in each group.The DR rat model was established by feeding the rats in both the DR group and crocin group with a high glucose and high fat diet,along with intraperitoneal injection(IP)of streptozotocin.Crocin IP was administered to the rats in the crocin group,whereas the rats in the DR group and control group received an equivalent dosage of saline IP for 12 weeks.A comparison was made among the three groups regarding retinal thickness,vascular permeability,expression of TLR4/My D88/NF-κB pathway protein,levels of inflammatory factors,and levels of Bcl-2,Bax,and Bcl-2/Bax.Results:The DR group and crocins group exhibited a lower retinal thickness compared to the control group,while the crocins group displayed a higher thickness than the DR group.The DR group and crocins group had higher retinal vascular permeability than the control group,and the crocins group had lower retinal vascular permeability than the DR group(P<0.05).TLR4,My D88,and P-NF-κB relative expressions were higher in the DR and crocin groups than in the control group,whereas TLR4,My D88,and P-NF-κB relative expressions were lower in the crocin group than in the DR group(P<0.05).The DR group and crocin group exhibited elevated levels of inflammatory cytokines compared to the control group,while the crocin group displayed decreased levels in comparison to the DR group(P<0.05).The DR group and crocin group exhibited lower levels of Bcl-2 and Bcl-2/Bax compared to the control group,whereas the control group displayed higher levels of Bax.The crocin group exhibited elevated levels of Bcl-2 and Bcl-2/Bax compared to the DR group,whereas the DR group displayed diminished levels of Bax(P<0.05).Conclusion:Crocin has the potential to enhance the retinal thickness and vascular permeability of DR rats,and the inhibition of the TLR4/My D88/NF-κB pathway by crocin could play a crucial role in impeding the advancement of DR.

枸橼酸咖啡因联合苯巴比妥对早产儿脑损伤患儿NF-κB、TLR2的影响

目的探讨枸橼酸咖啡因联合苯巴比妥对早产儿脑损伤患儿核转录因子κB(NF-κB)、Toll受体2(TLR2)的影响。方法在患儿监护人同意前提下选取2018年4月至2021年4月在南通大学第二附属医院出生的183例早产儿脑损伤患儿为研究对象,数字表法分为苯巴比妥治疗的单药组(91例)及枸橼酸咖啡因、苯巴比妥联合治疗的联合组(92例)。对比2组患儿神经行为能力,检测神经损伤标志物、炎性因子及NF-κB、TLR2水平,分析临床疗效。结果治疗后,联合组一般反应、行为能力、被动肌张力、主动肌张力、原始反射评分高于单药组(P<0.05);治疗后,血清髓鞘碱性蛋白(MBP)、神经元特异性烯醇化酶(NSE)、S100β蛋白(S100β)、NF-κB、TLR2、白介素6(IL-6)、超敏C反应蛋白(hs-CRP)水平降低,且联合组以上指标的水平低于单药组(P<0.05)。联合组临床治疗总有效率高于单药组(P<0.05)。结论枸橼酸咖啡因、苯巴比妥联合应用有效提高早产儿脑损伤患儿临床疗效,改善患儿神经行为,调控NF-κB、TLR2水平,抑制炎性反应,减轻脑损伤程度,疗效显著。