核因子κB、肿瘤坏死因子α在宫颈癌中的表达情况探究

目的探讨核因子κB(NF-κB)、肿瘤坏死因子α(TNFα)在宫颈癌中的表达情况。方法对40例正常宫颈,12例CINⅠ,11例CINⅡ~Ⅲ和61例宫颈鳞癌进行免疫组化SABC法检测,观察NF-κB、TNFα的表达情况,探究两者的表达水平与宫颈癌的内在关系。结果宫颈癌组NF-κB、TNFα的阳性表达率较其余3个组别的表达差异有统计学意义(P<0.05),而CIN I、CINⅡ~Ⅲ与正常宫颈组织的NF-κB、TNFα表达组间差异无统计学意义(P>0.05)。TNFα、NF-κB在宫颈鳞癌组织中的表达呈显著正相关(r=0.66,P<0.05)。结论 NF-KB、TNFα的过度激活在宫颈癌的发生、发展中可能起重要作用,值得临床上进一步研究。     

NF-κB、MMP-9在宫颈癌组织中的表达及意义

目的观察核转录因子-κB(nuclear factor kappa-B,NF-κB)和MMP-9(matrix metalloproteinase,MMP-9)在宫颈癌组织中的表达及临床意义。方法采用免疫组织化学SP法检测80例宫颈癌(CSES)、70例子宫颈上皮内瘤样变(CIN)及50例正常子宫颈组织(NCE)NF-κB和MMP-9蛋白表达水平,分析其与临床病理特征的关系。结果在CSES、CIN及NCE中,NF-κB蛋白阳性表达率分别为72.5%、18.6%、0%,MMP-9蛋白阳性表达率分别为68.8%、15.7%、0;即CSES中NF-κB和MMP-9蛋白表达显著高于CIN和NCE,差异具有统计学意义(P<0.05)。NF-κB、MMP-9表达与宫颈癌不同的病理分级、临床分期和淋巴结转移状况有关(P<0.05),与患者年龄和组织学类型无关,且二者表达呈明显正相关(r=0.872,P<0.05)。结论宫颈癌患者存在NF-κB和MMP-9的过表达,其对宫颈癌的发生发展、浸润和转移具有重要意义,可作为宫颈癌诊断及判断预后的标志物。     

Toll样受体4和9基因在肺纤维化大鼠的表达和意义

目的:观察Toll样受体9(TLR-9)及Toll样受体4(TLR-4)基因在肺成纤维细胞大鼠的动态演变及其在炎性反应中的作用.方法:博莱霉素诱导实验性大鼠肺纤维化模型,采用微波改良Masson染色、VG染色法、qRT-PCR等方法,观察实验性大鼠肺纤维化的发病过程及其肺组织中TLR-9和TLR-4基因表达水平的动态变化及其下游分子NF-KB和IFN-γ的表达.结果:实验性大鼠肺纤维化发病过程中,呈现典型的肺泡炎(7d)、纤维组织增生(14d)及稳定的肺纤维化(28~56d)等表现;与对照组相比,TLR-9、TLR-4、NF-KB和IFN-γ显著高于对照组,差异具有统计学意义(P<0.01);TLR-9与NF-KB和IFN-γ表达呈正相关(r=0.91、r=0.88,P<0.01);TLR-4与NF-KB和IFN-γ表达也呈正相关(r=0.96、r=0.98,P<0.01).结论:TLR-9和TLR-4可能参与肺纤维化的发生发展及调控大鼠肺成纤维的炎症反应.     

Toll样受体4促进宫颈上皮内瘤变进展的机制研究

目的探讨Toll样受体4(TLR4)在正常宫颈组织、宫颈上皮内瘤变(CIN)组织、宫颈癌中的表达情况,分析TLR4与CIN及宫颈癌的相关性,并研究其对临床诊断的影响及评价。方法将2018年4月至12月期间来我院进行宫颈检查女性,随机选取正常宫颈组织6例,CIN 17例以及宫颈癌7例,检测3组TLR4阳性表达情况,分析影响其表达的相关因素。结果 TLR4在正常宫颈组织、CIN以及宫颈癌组织中的表达强度呈递增升高,差异有统计学意义(P<0.05);TLR4的表达与患者年龄及细胞分化程度无明显关系(P>0.05);与肿瘤分期和人乳头瘤病毒(HPV)感染具有明显的相关性,差异有统计学意义(P<0.05)。结论 TLR4在正常宫颈组织、CIN、宫颈癌组织中的表达逐渐升高,TLR4的表达与HPV感染、CIN及宫颈癌发病具有相关性,能够为临床疾病的诊疗情况提供较优的参考价值。     

核转录因子NF-κB在子宫颈癌中的表达及其临床意义

目的：本研究通过检测核转录因子NF-κB在人宫颈癌中的表达,探讨其在宫颈癌发生、进展过程中的作用及临床意义。方法：应用免疫组织化学染色方法（S-P）检测宫颈癌组织42例、CIN 20例及正常宫颈10例中NF-κB表达水平。结果：宫颈癌组织中NF-kB蛋白阳性表达率为74.1%（30/42）,CIN组织中NF-kB蛋白阳性表达率为15%(3/20),正常组织中则无NF-kB表达,宫颈癌中NF-κB表达水平高于CIN与正常宫颈组织（P<0.05）;CIN中NF-κB表达水平与正常宫颈组织间无统计学差异（P＞0.05）。在宫颈癌中,NF-κB表达在低分化,与患者年龄、病理类型无关（P＞0.05）,NF-κB不仅高表达于低分化宫颈癌,而且随着临床分期增高、出现淋巴转移等,其表达增强。结论：NF-κB不是导致子宫颈癌前病变的主要机制,可能只是表达量的积累过程。而NF-κB异常表达及激活可能是子宫颈癌发生、进展过程中的重要机制之一。     

蚤休薯蓣皂苷对内皮细胞的保护作用涉及TLR4/NF-κB途径

目的观察蚤休薯蓣皂苷(Rhizome Dioscin,RD)对氧化性低密度脂蛋白(ox-LDL)诱导人脐静脉内皮细胞(HU-VEC)损伤对细胞生长、炎症反应Toll样受体4(TLR4)和细胞核因子κB(nuclear factor kappa B,NF-κB)表达的影响,探讨RD抗动脉粥样硬化(AS)的作用机制。方法体外培养HUVEC,RD 3个浓度预处理,加入ox-LDL,运用流式细胞术PI染色检测细胞周期,激光共聚焦显微镜JC-1染色检测线粒体电位(ΔΨm),Western blot检测TLR4和NF-κB蛋白的表达。结果与正常对照组相比,经ox-LDL损伤后HU-VEC细胞周期阻滞在G0/G1期,表现为G0/G1期细胞比例明显升高(P<0.01),S期细胞比例明显下降(P<0.01);细胞ΔΨm明显下降;TLR4和NF-κB的表达与正常组比较均升高。RD预处理能抑制ox-LDL诱导的细胞生长停滞,使细胞周期S期细胞比例增加(P<0.01),ΔΨm上升,TLR4和NF-κB的表达与损伤组相比均下降。结论抑制TLR4/NF-κB信号转导途径可能是RD抗内皮细胞氧化损伤、阻止经线粒体途径的细胞生长停滞的可能机制之一。     

TLR9和NF-κBp65在大鼠溃疡性结肠炎模型结肠组织中的表达

目的观察TLR9(Toll样受体9)、NF-κBp65(核因子-κBp65)在大鼠溃疡性结肠炎(UC)模型结肠组织中的表达。方法应用复合法(2,4-二硝基氯苯+乙酸)制备大鼠UC模型,观察和评估结肠黏膜的大体形态和组织学变化。以免疫组化Elivision法检测TLR9、NF-κBp65的表达。结果造模后可见大鼠肠黏膜有大量炎性细胞浸润,累及黏膜下层和固有层。与正常对照组相比,模型组结肠组织TLR9、NF-κBp65表达显著升高(P0.01)。结论 TLR9、NF-κBp65在UC发病机制中具有重要作用。     

Endoglin与血管内皮生长因子和血管内皮生长因子受体2及基质金属蛋白酶9联合表达与宫颈癌血管生成的关系

目的 探讨Endoglin、血管内皮生长因子(VEGF)、血管内皮生长因子受体2(KDR)及基质金属蛋白酶9(MMP-9)联合表达在宫颈癌局部血管生成中的作用.方法 采用免疫组织化学SP法检测15例正常宫颈上皮(NCE)、18例宫颈上皮内瘤变(CIN)和75例宫颈癌(ICC)组织Endoglin、VEGF、KDR及MMP-9表达,并检测CD34标记的微血管密度(MVD).结果 Endoglin、VEGF、KDR和MMP-9在NCE、CIN及ICC组的阳性率分别为6.7%、38.9%及64.0%;26.7%、61.1%及77.3%;13.3%、61.1%及69.3%和20.0%、55.6%及81.3%,从NCE到CIN再到ICC组,Endoglin、VEGF、KDR及MMP-9的阳性率均显著升高(P<0.05).宫颈癌组织MVD均分别与Endoglin(r=0.499,P<0.01)、VEGF(r=0.602,P<0.01)、KDR(r=0.331,P<0.01)及MMP-9(r=0.287,P<0.05)表达显著正相关,并且Endogin、VEGF、KDR及MMP-9同时均阳性表达者,其MVD升高更为显著(P<0.01).结论 Endoglin、VEGF、KDR及MMP-9联合表达在上调宫颈癌局部肿瘤血管生成中起重要作用,4个指标均过度表达,血管生成能力显著增强.     

辛伐他汀对1型糖尿病大鼠的肾脏保护作用及对肾组织Toll样受体4和核因子-κB表达的影响

目的观察辛伐他汀对糖尿病大鼠的肾脏保护作用及对肾组织Toll样受体(TLR)4和核因子(NF)-κB表达的影响。方法将54只SD大鼠分为正常对照组、糖尿病组和辛伐他汀治疗组。实验第4、8、12周,测定各组大鼠的血糖、肾脏肥大指数和尿蛋白定量(24 h),免疫组织化学法和荧光定量聚合酶链反应(FQ-PCR)法分别检测肾组织TLR4和NF-κB蛋白和mRNA的表达;电镜下观察第12周肾组织的超微结构变化。结果与正常对照组比,糖尿病组的血糖、肾脏肥大指数和尿蛋白定量(24 h)显著增加(P<0.05),肾组织TLR4和NF-κB蛋白和mRNA的表达显著增高(P<0.05),肾小球基底膜显著增厚,系膜和系膜细胞显著增生;辛伐他汀治疗组的上述各项指标较糖尿病组均明显降低(P<0.05),但较正常对照组仍有增高趋势(P<0.05),病理改变亦明显减轻,但仍达不到正常对照水平。结论辛伐他汀对糖尿病早期肾损伤有一定的保护作用,其机制可能是通过下调TLR4和NF-κB的表达,抑制炎症反应,降低尿蛋白而减轻糖尿病肾损害。     

脂氧素A4受体激动剂及白介素-1β在Toll样受体2/髓样分化因子88/核因子-κB信号通路对哮喘小鼠气道炎症的调控作用

目的:研究脂氧素(LX)A4受体激动剂及白介素-1β抗体在Toll样受体2(TLR2)/髓样分化因子88(MyD88)/核因子-κB（NF-κB)信号通路中对哮喘小鼠气道炎症的调控机制。方法:以卵清蛋白(OVA)致敏激发法制备小鼠哮喘模型,60只二级雄性Balb/c小鼠随机分为6组,分别给予脂氧素A4受体激动剂(BML-111)、白介素(IL)-1β抗体、BML-111载体、兔IgG治疗。末次激发后留取血清和肺组织标本,光镜下观察支气管周围炎症浸润情况;测定血清IL-1β、IL-4、IL-8、干扰素(IFN)-γ水平;检测肺组织TLR2、MyD88、NF-κB的表达。结果:OVA致敏的小鼠血清IL-1β、IL-4、IL-8水平升高,IFN-γ浓度降低; OVA致敏导致明显的支气管周围炎症细胞浸润,使支气管炎症分级指数有明显提高;BML-111或IL-1β抗体治疗均能明显阻止上述OVA介导的炎症变化(χ2=28.14,P<0.01)。OVA致敏使肺组织TLR2、MyD88 表达及NF-κB活性升高,BML-111及抗IL-1β抗体可抑制由OVA引发的上述表达变化。结论:OVA可诱导产生气道炎症,这一作用可能受TLR2/MyD88/NF-κB信号通路调控,IL-1β在这一过程中起了至关重要的作用。BML-111和IL-1β抗体可能通过对TLR2/MyD88/NF-κB信号通路的负调控而实现抑制OVA诱发的呼吸道炎症的作用。     

TLR2、TLR4与类风湿关节炎

Toll样受体（TLRs）是天然免疫系统中特异性的Ⅰ型跨膜受体及病原模式识别受体,在急性炎症反应、细胞信号转导和细胞凋亡中起重要作用。TLRs通过选择性的识别病原体中病原相关分子模式的保守结构及某些内源性配体,触发髓样细胞分化因子88（MyD88）依赖性和非依赖性途径,导致基因编码的的促炎性细胞因子和趋化因子的激活,从而诱发局部的炎症,并通过上调抗原递呈细胞表面的共刺激分子,诱导T、B淋巴细胞分化,激动后继的适应性免疫应答。类风湿关节炎（RA ）主要表现为滑膜组织炎性增生和炎症细胞浸润最终导致组织破坏和功能障碍。与T细胞过度活化以及B细胞的过度刺激致大量自身抗体产生有关。研究发现,细胞表面的TLRs,尤其是TLR2和TLR4,在类风湿关节炎疾病的发生发展中起重要作用。该文就TLR2、TLR4在RA发病中的作用作一综述,旨在为后期药物干预提供一定的理论依据。     

RNA recognition via TLR7 and TLR8

In this chapter we focus on immunorecognition of RNA by two members of the family of Toll-like receptors (TLRs), TLR7, and TLR8. While any long single-stranded RNA is readily recognized by both TLR7 and TLR8, sequencedependent activation of TLR7 and TLR8 becomes more evident when using short RNA oligonucleotides. RNA oligonucleotides containing sequence motifs for TLR7 and TLR8 are termed is RNA (immunostimulatory RNA). Moreover, short doublestranded RNA oligonucleotides as used for siRNA (short interfering RNA) containing such sequences function primarily as ligands for TLR7 but not TLR8. Even in the presence of appropriate sequence motifs, RNA is not detected by TLR7 and TLR8 when certain chemical modifications are present. Both immunological recognition and ignorance are relevant for the development of RNA-based therapeutics, depending on the clinical setting for which they are developed.     

TLR2 and TLR4 expression during bacterial infections

The family of the toll-like receptors comprises a minimum of 10 members identified in humans so far. These transmembrane receptors act as important signaling intermediates between the host and the invading pathogens. The following review describes the complexities encountered by researchers studying toll-like receptor (TLR) expression changes during bacterial infections. Mutations in some of the TLRs, most prominently TLR4 and TLR2, have been associated with increased susceptibility to infectious diseases. While it is tempting to correct the phenotypic effect of such mutations, in vitro and in vivo research has shown that TLR activity and function comprises a complex regulatory network. Heterodimer formation, synergy, and cross-tolerance have previously been described. More recently, interdependence of TLR2 and TLR4 expression has been identified. In addition, TLR expression follows a specific timeline that may be dependent on the invading pathogen. Lastly, mutations in invading pathogens have been shown to alter the expression profile of TLR2 and TLR4, indicating that therapies against bacterial pathogens will have to target multiple TLRs. Despite the complexities involved in TLR function, the significant progress made in our understanding of the role these proteins play in human diseases also indicates their potential value as therapeutic agents.     

TLR signalling and association of TLR polymorphism with cardiovascular diseases

Toll-Like receptors (TLRs) are the primary receptors of innate immunity. Considerable evidences have shown that innate immune defence interaction with pro-inflammatory pathways could be through TLRs that in turn leads to development of inflammatory diseases. These TLRs are present on various tissues and cells of cardiovascular system. Previous studies involving SNPs analysis of TLRs demonstrated that TLRs are involved in development and progression of diseases like atherosclerosis, cardiac dysfunction in sepsis and congestive heart failure.In this review, we aimed to bring together the studies which have been conducted previously to establish a link between TLR polymorphism in context to development of cardiovascular diseases (CVD).     

黄芩苷下调沙眼衣原体感染小鼠TLR2和TLR4基因表达的研究

目的:研究黄芩苷对沙眼衣原体(Chlamydia trachomatis,Ct)感染小鼠宫颈组织Toll样受体2和Toll样受体4(TLR2,TLR4)基因表达的影响。方法:6~8周龄雌性C57BL/6小鼠,分为5组:阿奇霉素组,模型组,黄芩苷高、中、低剂量组。通过阴道接种Ct感染小鼠,建立Ct感染小鼠生殖道的动物模型。接种前7 d皮下注射黄体酮以增加小鼠对Ct感染的敏感性。通过比较各组小鼠阴道分泌物中的Ct数量,评价黄芩苷抗沙眼衣原体感染的能力。感染27 d之后,用RT-PCR法和Western Blot法检测宫颈组织TLR2和TLR4基因表达情况。结果:在阴道接种Ct后第3天和第6天,模型组小鼠阴道分泌物中Ct数量显著升高。通过黄芩苷治疗后,各药物组小鼠阴道分泌物中,Ct数量均明显少于模型组,黄芩苷高剂量组和阿奇霉素组显著性降低。同时,黄芩苷还可明显抑制各组宫颈组织内TLR2和TLR4基因的表达。结论:黄芩苷可有效地抑制Ct感染的小鼠宫颈组织TLR2和TLR4的高表达,这可能是黄芩苷抗Ct感染的作用机制之一。     

TLR based therapeutics

Toll-like receptors (TLRs) play a crucial role in innate immune responses to infection. Binding of agonists to TLRs promotes maturation of antigen presenting cells, such as dendritic cells, which in turn directs the induction of adaptive immune responses. For this reason TLR agonists are being exploited as vaccine adjuvants for infectious disease or cancer and as therapeutics against tumors. However TLR agonists also promote inflammatory cytokine production and have a pathogenic role in many diseases with an inflammatory basis, including autoimmune diseases. Consequently, antibodies to TLRs and inhibitors of TLR signalling pathways have considerable potential as therapeutics for inflammatory disorders. Some have shown to be efficacious in pre-clinical models, and have now entered clinical trials.     

Curcumin attenuates Concanavalin A-induced liver injury in mice by inhibition of Toll-like receptor (TLR) 2, TLR4 and TLR9 expression

Curcumin has antiviral, antioxidant, and anti-inflammatory properties. However, the hepatoprotective effects and molecular mechanisms of curcumin on acute liver injury have not been carefully examined. The aims of this study were to examine the anti-inflammatory effect of curcumin on Concanavalin A (Con A) induced hepatitis, and to elucidate its underlying molecular mechanisms in mice. Mice received curcumin (200 mg/kg body weight) by gavage before Con A intravenous administration. We found that curcumin pretreatment was able to significantly reduce the elevated plasma aminotransferase levels and liver necrosis in Con A-induced hepatitis. Also, curcumin pretreatment reduced intrahepatic expression of genes encoding pro-inflammatory molecules such as tumor necrosis factor α (TNF-α) and interferon γ (IFN-γ) as compared with the vehicle controls, but augmented anti-inflammatory cytokine interleukin 10 (IL-10) by enzyme linked immunosorbent assay (ELISA). Furthermore, the expression levels of Toll-like receptor (TLR) 2, TLR4 and TLR9 mRNA or protein in liver tissues were significantly lowered by curcumin treatment. Curcumin pretreatment did not affect hepatic Kupffer cell numbers after Con A injection. These results suggest that curcumin pretreatment protects against T cell-mediated hepatitis in mice. The beneficial effect of curcumin may be partly mediated by inhibiting the expression levels of TLR2, TLR4 and TLR9 in the liver.     

TLR2 and TLR4 expression after kidney ischemia and reperfusion injury in mice treated with FTY720

Ischemia and reperfusion injury (IR) is an antigen independent inflammatory process that causes tissue damage. After IR, kidneys up-regulate leukocyte adhesion molecules and toll-like receptors (TLRs). Moreover, injured kidneys can also secrete factors (i.e. heat shock protein) which bind to TLRs and trigger intracellular events culminating with the increase in the gene expression of inflammatory cytokines. FTY720 is an immunomodulatory compound and protects at least in part kidneys submitted to IR. The mechanisms associated with FTY720's beneficial effects on kidneys after IR remain elusive. We investigated whether FTY720 administration in mice submitted to kidney IR is associated with modulation of TLR2 and TLR4 expression. C57BL/6 mice submitted to 30min of renal pedicles clamp were evaluated for serum parameters (creatinine, urea and nitric oxide), kidney histology, spleen and kidney infiltrating cells expression of TLR2 and TLR4, resident kidney cells expression of TLR2 and TLR4 and IL-6 protein expression in kidney. FTY720-treated mice presented decrease in serum creatinine, urea and nitric oxide, diminished expression of TLR2 and TLR4 both in spleen and kidney infiltrating cells, and reduced kidney IL-6 protein expression in comparison with IR non-treated mice. However, acute tubular necrosis was present both in IR non-treated and IR+FTY720-treated groups. Also, FTY720 did not prevent TLR2 and TLR4 expression in kidney resident cells. In conclusion, FTY720 can promote kidney function recovery after IR by reducing the inflammatory process. Further studies are needed in order to establish whether TLR2 and TLR4 down regulation should be therapeutically addressed as protective targets of renal function and structure after IR.