布地奈德混悬液对哮喘模型小鼠肺组织TLR4/MyD88/NF-κB通路的影响

摘要 目的：探究布地奈德混悬液对哮喘模型小鼠肺组织TOLL样受体4（toll-like receptor 4,TLR4）/髓样分化因子88（myeloid differentiation factor 88,MyD88）/核因子κB（nuclear factor-κB,NF-κB）通路的影响。方法：使用4 %鸡蛋清白蛋白与2 %的Al（OH3）共同致敏小鼠,建立咳嗽变异性哮喘小鼠模型40只,将模型大鼠分别使用低、中、高剂量（0.2、1.0、2.0 g/kg）布地奈德混悬液和孟鲁司特钠进行干预,1次/日连续干预14 d,于干预14 d时采集小鼠的支气管肺泡灌注液（bronchoalveolar lavage fluid,BALF）、气管及肺组织,对各组BALF中的白细胞（white blood cell,WBC）、嗜酸性粒细胞、血清γ干扰素（interferon-γ,IFN-γ）、白细胞介素-1β（interleukin-1β,Il-1β）、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平差异开展比较,对各组小鼠肺组织黏膜上皮增生程度评分、炎症细胞浸润程度评分、病变总评分差异,以及TLR4、MyD88、p65蛋白表达差异进行分析。结果：分析显示,布地奈德混悬液能够显著降低哮喘模型小鼠BALF中白细胞及嗜酸性粒细胞数量,同时还能够改善小鼠气管和支气管黏膜上皮增生与肺组织炎症细胞浸润状态,且干预后小鼠肺组织中的TLR4、MyD88、p65蛋白表达水平出现了明显的降低。结论：布地奈德混悬液对改善小鼠哮喘效果较好,其作用机制可能与该药能够调节TLR4、MyD88、p65蛋白表达,进而影响炎症和免疫反应进程有关。     

胀果甘草多糖GiP-B1通过TLR4/MyD88/NF-κB言号通路激活巨噬细胞RAW 264.7

为研究胀果甘草多糖GiP-B1通过TLR4/MyD88/NF-κB信号通路对巨噬细胞RAW 264.7免疫功能的影响,体外以TLR4干扰RNA (TLR4-siRNA)转染巨噬细胞RAW 264.724 h,用100 μg/mL GiP-B1处理转染和未转染细胞,12 h后分别采用CCK-8和ELISA法检测GiP-B...     

基于TLR4/MyD88/NF-κB通路探讨美洲大蠊虫粉对脊髓损伤大鼠的保护作用

目的 探讨美洲大蠊虫粉对脊髓损伤大鼠的保护作用和可能的机制。方法 将48只SD大鼠随机分为假手术组、盐水组、美洲大蠊虫粉组、TOLL样受体4(toll-like receptors, TLR4)抑制剂组。除假手术组外其余3组均构建大鼠脊髓半横断损伤模型。术后假手术组不做治疗,盐水组与美洲大蠊虫粉组分别给予生理盐水和美洲大蠊虫粉(630 mg/kg)灌胃处理,TLR4抑制剂组给予TLR4抑制剂(3 mg/kg)腹腔注射处理。术后1、3、7、14 d运用BBB评分评估大鼠后肢运动功能,苏木素-伊红(HE)染色观察脊髓组织病理改变,免疫组化观察神经元数目变化,酶联免疫法检测炎性因子IL-1、IL-6、IL-10和TNF-α表达,Western Blot检测TLR4、髓样分化因子(myeloid differentiation factor 88,MyD88)和NF-κB p65表达。结果 与假手术组相比,盐水组BBB评分、神经元数目显著下降,而病理损伤程度、IL-1、IL-6、TNF-α、TLR4、MyD88、NF-κB p65水平显著增加(P<0.05),与盐水组相比,美洲大蠊虫粉组...     

TLR/NF-κB信号通路与肺部疾病

Toll样受体(Toll like receptor,TLR)是一种重要的模式识别受体,核转录因子-κB(nuclear factor-κB,NF-κB)处于TLR下游信号通路中的关键位置,当TLR受到病原微生物刺激后,激活NF-κB,诱导炎症因子释放,启动固有免疫。但TLR/NF-κB信号通路过度激活,有可能导致炎症反应失控。本文将介绍TLR/NF-κB信号通路及其在肺部炎症疾病例如急性肺损伤、慢性阻塞性肺疾病、肺癌、哮喘等发生发展中的作用。     

过表达miR-31对结肠炎模型小鼠TLR4/NF-κB信号通路及凋亡蛋白的调控

目的: 探讨miR-31对DSS诱发结肠炎小鼠TLR4/NF-κB信号通路和凋亡相关蛋白的影响。方法: ①小鼠结肠炎实验:用1%葡聚糖硫酸钠(DSS)诱发小鼠溃疡性结肠炎(UC)。14只FVB非转基因小鼠随机分为control组(n=6),DSS组(n=8),16只FVB miR-31转基因小鼠随机分为miR-31过表达组(n=8),miR-31过表达+DSS 组(n=8),DSS溶于水后通过饮水给予小鼠。DSS组和miR-31+DSS组第一周饮用1%DSS水,第二周饮用正常无菌水,第三周饮用1%DSS水,如此5周后造模完成,之后留取小鼠的结肠组织,通过Western blot和IHC检测小鼠结肠组织NF-κB p65、TLR4、Bax、Bcl-2蛋白的表达;TUNEL检测小鼠结肠组织细胞凋亡。②细胞培养实验:在人结肠上皮细胞系HCT 116细胞中通过脂质体转染的方法转染miR-31 mimic和inhibitor,使miR-31过表达或敲低,每组均进行三次重复,48 h后收取细胞,通过Western blot检测NF-κB p65、TLR4蛋白的表达。结果: ①动物实验中,与control组相比,小鼠结肠组织中DSS组和miR-31过表达组NF-κB p65、TLR4蛋白表达水平和凋亡细胞指数均显著升高(P＜0.05或P＜0.01),Bcl-2/Bax比值显著降低(P＜0.05或P＜0.01);且与DSS组相比,miR-31+DSS组NF-κB p65、TLR4蛋白表达水平和凋亡细胞指数也显著升高(P＜0.01),Bcl-2/Bax比值显著降低(P＜0.01)。②细胞实验中,与control组相比, HCT 116细胞过表达miR-31组的NF-κB p65、TLR4蛋白表达水平均显著升高(P＜0.05或P＜0.01),敲低miR-31组的NF-κB p65、TLR4蛋白表达水平下降(P＜0.05)。结论: miR-31通过促进TLR4/NF-κB信号通路和介导肠上皮细胞凋亡促进结肠炎的发展。     

Eupatilin alleviates inflammatory response after subarachnoid hemorrhage by inhibition of TLR4/MyD88/NF-κB axis

Early brain injury (EBI) is associated with the adverse prognosis of subarachnoid hemorrhage (SAH) patients. The key bioactive component of the Chinese herbal medicine Artemisia asiatica Nakai (Asteraceae) is eupatilin. Recent research reports that eupatilin suppresses inflammatory responses induced by intracranial hemorrhage. This work is performed to validate whether eupatilin can attenuate EBI and deciphers its mechanism. A SAH rat model was established by intravascular perforation in vivo. At 6 h after SAH in rats, 10 mg/kg eupatilin was injected into the rats via the caudal vein. A Sham group was set as the control. In vitro, BV2 microglia was treated with 10 μM Oxyhemoglobin (OxyHb) for 24 h, followed by 50 μM eupatilin treatment for 24 h. The SAH grade, brain water content, neurological score, and blood-brain barrier (BBB) permeability of the rats were measured 24 h later. The content of proinflammatory factors was detected via enzyme-linked immunosorbent assay. Western blot analysis was conducted to analyze the expression levels of TLR4/MyD88/NF-κB pathway-associated proteins. In vivo, eupatilin administration alleviated neurological injury, and decreased brain edema and BBB injury after SAH in rats. Eupatilin markedly reduced the levels of interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α), and suppressed the expression levels of MyD88, TLR4, and p-NF-κB p65 in the SAH rats' cerebral tissues. Eupatilin treatment also reduced the levels of IL-1β, IL-6, and TNF-α, and repressed the expression levels of MyD88, TLR4, and p-NF-κB p65 in OxyHb-induced BV2 microglia. Additionally, pyrrolidine dithiocarbamate or resatorvid enhanced the suppressive effects of eupatilin on OxyHb-induced inflammatory responses in BV2 microglia. Eupatilin ameliorates SAH-induced EBI via modulating the TLR4/MyD88/NF-κB pathway in rat model.     

血红素加氧酶-1对急性重症胰腺炎相关肺损伤TLR4/NF-κB通路的影响

目的:探讨血红素加氧酶-1(HO-1)对急性重症胰腺炎相关肺损伤(PALI)Toll样受体-4(TLR4)/核因子-κB(NF-κB)信号传导通路的影响。方法:32只SD大鼠随机分为Sham组、PALI组、HO-1促进剂组、HO-1抑制剂组,每组8只。PALI组经胆胰管注入牛磺胆酸钠制备急性重症胰腺炎(ANP)动物模型。Sham组胆胰管内不注入牛磺胆酸钠,其余操作同PALI组。HO-1促进剂组于造模后30 min经腹腔注射牛血晶素75μg/kg;HO-1抑制剂组于造模后30 min经腹腔注射锌-原卟啉20μmol/kg。PALI组和Sham组均于造模后30 min经腹腔注射等量生理盐水。各组大鼠术后24 h,进行肺损伤学评分,统计肺湿/干重比值。检测大鼠术后24 h血清淀粉酶、TNF-α、IL-6、NGAL水平。检测大鼠术后24 h肺组织中TLR4、NF-κB p65蛋白表达。结果:PALI组肺损伤学评分、肺湿/干重比值、淀粉酶、TNF-α、IL-6、NGAL、TLR4、NF-κB p65明显高于Sham组;HO-1促进剂组肺损伤学评分、肺湿/干重比值、淀粉酶、TNF-α、IL-6、NGAL、TLR4、NF-κB p65明显低于PALI组;HO-1抑制剂组肺损伤学评分、肺湿/干重比值、淀粉酶、TNF-α、IL-6、NGAL、TLR4、NF-κBp65明显高于PALI组;差异均有统计学意义(P<0.05)。结论:HO-1能够通过抑制TLR4/NF-κB信号通路的激活,下调TNF-α、IL-6、NGAL等炎症因子的释放,从而发挥减轻急性重症胰腺炎相关肺损伤的作用。     

胰腺癌上调因子在卵巢癌中水平变化及其与TLR4/MyD88通路的关系

该文探讨了胰腺癌上调因子(PAUF)在卵巢癌中的表达水平,及其与TLR4/MyD88信号通路在卵巢癌中的关系。选取2014年10月至2016年5月在我院就诊的卵巢癌患者共217例,纳入卵巢癌组,另随机选取同期诊断为卵巢良性肿瘤的45例患者纳入卵巢良性肿瘤组和52例健康人群纳入对照组。采用免疫组织化学染色法检测受试者卵巢组织中的PAUF、TLR4和MyD88表达水平。比较三组PAUF、TLR4和MyD88的表达水平;分析PAUF和TLR4、MyD88之间存在的相关性;分析卵巢癌患者中影响PAUF、TLR4和MyD88表达水平的因素。结果显示,卵巢癌患者组织中的PAUF、TLR4和MyD88水平均显著高于良性肿瘤组和对照组(P0.05);卵巢癌组织中PAUF与TLR4表达水平呈正相关(r=0.521,P0.001),且PAUF与MyD88表达也成正相关(r=0.581, P0.001);卵巢癌组织PAUF水平与肿瘤种类、FIGO分期和肿瘤分化程度有关(P0.05),但与患者年龄无关(P0.05);而癌组织中TLR4水平只与肿瘤的分化程度有关(P0.05), MyD88表达水平只与癌症种类有关(P0.05)。由此说明,卵巢癌患者组织中的PAUF表达水平显著升高,与TLR4和MyD88表达成正相关。PAUF可能通过参与TLR4/MyD88信号通路,参与了卵巢癌的发生发展。     

Xiao-Chai-Hu-Tang ameliorates tumor growth in cancer comorbid depressive symptoms via modulating gut microbiota-mediated TLR4/MyD88/NF-κB signaling pathway

BackgroundDepressive symptoms are thought to promote cancer development and depressive remission has been reported to be effective for defeating cancer. The herbal formula Xiao-Chai-Hu-Tang (XCHT), that has an anti-depressive efficacy, has been widely utilized in China. However, its anti-cancer effect and underlying mechanisms remain unclear.PurposeThe present study aims to investigate the effects of XCHT on the depression-associated tumor and its potential mechanisms.MethodsA placebo-controlled trial was conducted in cancer patients comorbid with depressive symptoms to evaluate the effects of XCHT on depressive scales, tumor-related immune indicators, and gut microbial composition. A xenografted colorectal cancer (CRC) mouse model exposure to chronic restraint stress (CRS) was established to examine XCHT effects on tumorigenesis in vivo. Further, by manipulating gut bacteria with fecal microbial transplantation (FMT) or antibiotics-induced bacterial elimination in CRS-associated xenografted model, gut microbiota-mediated anti-tumor mechanism was explored.ResultsIn cancer patients comorbid with depressive symptoms, XCHT showed substantial effects on improvement of depressive scales, system inflammatory levels and gut dysbiosis. In vivo, XCHT inhibited tumor growth and prolonged survival time in addition to showing anti-depressive effect. Similarly, in our clinical trial, XCHT partially reversed gut dysbiosis, particularly through reducing abundances of Parabacteroides, Blautia and Ruminococcaceae bacterium. Manipulation of gut bacteria in CRS-associated xenografted model further proved that the inhibition of XCHT on tumor progression was mediated by gut microbiota and that the underlying mechanism involves in downregulation of TLR4/MyD88/NF-κB signaling.ConclusionsWe demonstrated that gut microbiota mediates the anti-tumor action of the formula XCHT in cancer patients and models that were comorbid with depressive symptoms. This study implies a novel clinical significance of anti-depressive herbal medicine in the cancer treatment and clarifies the important role of gut microbiota in treating cancer accompanied by depressive symptoms.     

七味白术散对糖尿病大鼠肠道屏障和TLR4/NF-κB信号通路的影响

【背景】七味白术散可用于治疗气阴两虚所致的消渴病,尤其近些年来在改善糖尿病胰岛素抵抗方面得到广泛使用。【目的】观察七味白术散对糖尿病大鼠肠道屏障和炎症因子的影响,并探讨其作用机制。【方法】60只无特定病原体(specific pathogen free, SPF)级SD雄性大鼠随机分为空白组、模型组、二甲双胍组和七味白术散高、中、低剂量组,每组10只。腹腔注射链脲佐菌素(streptozotocin, STZ)的方法建立糖尿病大鼠模型,空白组大鼠给予正常饮食,模型组、二甲双胍组和七味白术散高、中、低剂量组给予高糖高脂饮食。造模成功后分别用二甲双胍和七味白术散灌胃处理,空白组和模型组生理盐水灌胃。观察治疗前后大鼠的一般生存状况;药物干预4周后,全自动生化分析仪检测空腹血糖(fasting blood glucose, FBG)、空腹胰岛素(fasting insulin, FINS)、脂多糖(lipopolysaccharide, LPS);酶联免疫吸附试验(enzyme-linked immunosorbent assay, ELISA)测定血清白细胞介素-6 (interleukin...     

基于TLR4MyD88信号通路观察粪菌移植对DSS诱导小鼠溃疡性结肠炎干预作用机制

摘要 目的：探讨粪菌移植（fecal microbiota transplantation,FMT）对葡聚糖硫酸钠（dextran sodium sulfate, DSS）诱导小鼠溃疡性结肠炎（ulcerative colitis,UC）干预作用及其炎症相关的分子机制。方法：30只昆明小鼠采用DSS喂养建立结肠炎小鼠模型并随机分为模型组、FMT低剂量组和FMT高剂量组,另取10只作为对照组。FMT低剂量组、FMT高剂量组自造模后第1 d开始分别给予8 g/kg、15 g/kg粪菌剂量灌肠处理,灌肠体积0.2 mL。对照组和模型组给予等体积生理盐水灌肠处理。分别在实验第1、7、14及21 d称量小鼠体重。实验结束后处死小鼠,取出结直肠,观察各组小鼠结直肠形态变化并通过HE染色观察病变程度。ELISA检测小鼠结直肠组织匀浆液上清中肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）、白细胞介素-6（interleukin 6, IL-6）、白细胞介素-4（interleukin 4, IL-4）和白细胞介素-10（interleukin 10, IL-10）的表达变化;免疫组化法和RT-qPCR分别检测结直肠组织中核因子κB-P65亚基 (nuclear factor-κB P65, NF-κB P65)蛋白和mRNA的表达情况;Western blotting检测结直肠组织中Toll 样受体4 (Toll-like receptor 4,TLR4)、髓样分化因子 88(myeloid differentiation factor 88,MyD88)的蛋白表达情况。结果：对照组相比,造模小鼠在第6 d起至第14 d体重明显减轻,差异具有统计学意义（P<0.05）;与模型组相比,FMT低、高剂量组小鼠第10 d起至第14 d体重明显减轻,FMT高剂量组小鼠体重升高更加明显,差异具有统计学意义（P<0.05）;与对照组相比,模型组小鼠结直肠组织中TNF-α、IL-6、NF-κB P65 mRNA和蛋白阳性表达、TLR4、MyD88 蛋白表达均显著升高(P<0.05),IL-4、IL-10表达显著下降（P<0.05）;FMT高剂量组IL-4、IL-10较模型组显著升高,其余指标均显著下降（P<0.05）。结论：FMT可通过抑制TLR4/MyD88/NF-κB信号通路缓解炎症反应,发挥对UC的治疗作用。     

运动干预下CSE/H2S调控TLR4/NF-κB信号通路对酒精性肝损伤的改善作用

胱硫醚-γ-裂解酶(cystathionine γ-lyase, CSE)是合成内源性H₂S的核心酶之一。CSE/H₂S体系可介导多种信号转导途径减轻机体炎性损伤。而有氧运动已被证实对机体免疫功能具促进作用,但是否通过CSE/H₂S体系介导炎性通路发挥效应,其机制有待深入研究。本研究旨在探讨有氧运动通过CSE/H₂S体系抑制TLR4/NF-κB信号通路对酒精性肝损伤的改善作用。选取3周龄健康雄性昆明(KM)种小鼠50只,随机分成酒精性脂肪肝病组(AFLD)[模型组(M)、有氧运动组(E)、有氧运动+NaHS组(EN)、有氧运动+PAG组(EP)]、空白组(K),每组10只。干预7周,解剖学观察发现,M组小鼠脂肪系数、脏器系数均高于K组(P<0.01)。ELISA酶联免疫结果显示,相比K组,M组谷草转氨酶、谷丙转氨酶水平增高,CSE活性下降,丙二醛含量上升,蛋白质羰基化程度上升及谷胱甘肽含量下降(P<0.01, P<0.05)。去蛋白质法检测发现,外周血H₂S含量升高(P<0.01)。HE染色显示,M组肝组织结构紊乱,呈现大量脂滴空泡样变,且胞核畸形位变。给予有氧运动干预及腹腔注射NaHS,可显著减轻肝质变及肝功能症状,提升血清H₂S含量和CSE活性(P<0.01)。而腹腔注射PAG可加剧酒精性肝损伤。免疫组织化学染色显示,相比M组,E、EN组TLR4、NF-κB、IL-1β阳性表达面积下降(P<0.01)。实时荧光定量PCR显示,相比M组,E组、EN组CSE、TLR4、NF-κB、IL-1β mRNA表达显著下降(P<0.01),EP组无显著差异(P>0.05)。Illumina高通量测序筛选肝组织炎症相关因子及关联分析显示,差异表达因子主要富集在NF-κB、TGF-β、TNF、TOLL样受体等信号通路,涉及肝组织细胞信号转导、凋亡抑制和免疫反应等,有氧运动及NaHS可降低炎症和免疫相关信号通路的富集。以上研究结果表明,有氧运动可促进小鼠肝组织中CSE/H₂S气体信号体系的表达,从而抑制TLR4/NF-κB通路促炎过程,拮抗小鼠酒精性肝损伤,且有氧运动结合外源性H₂S供体对TLR4/NF-κB的干预效果更佳。     

Long noncoding RNA ANRIL contributes to the development of ulcerative colitis by miR-323b-5p/TLR4/MyD88/NF-κB pathway

The aim of this study was to investigate the role and possible mechanism of long noncoding RNA ANRIL in the development of ulcerative colitis (UC). The expression of ANRIL in colonic mucosa tissues collected from the sigmoid colon of UC patients and healthy control was determined. Subsequently, fetal human cells (FHCs) were treated with lipopolysaccharide (LPS) to stimulate UC-caused inflammatory injury, followed by detection of the effects of suppression of ANRIL on cell viability, apoptosis and cytokines production in LPS-stimulated FHCs. Moreover, the regulatory relationship between ANRIL and miR-323b-5p as well as the target relationship between miR-323b-5p and TLR4 were investigated. Furthermore, the effects of ANRIL/miR-323b-5p axis on the activation of TLR4/MyD88/NF-κB pathway in LPS-stimulated FHCs were investigated. LncRNA ANRIL was highly expressed in colonic mucosa tissues of UC patients. In addition, LPS markedly induced cell injury in FHC cells (inhibited cell viability and promoted cell apoptosis and cytokine production). Suppression of ANRIL alleviated LPS-induced injury in FHC cells, which was achieved by negatively regulating miR-323b-5p. Moreover, miR-323b-5p negatively regulated TLR4 expression and TLR4 was a target of miR-323b-5p. Knockdown of TLR4 reversed the effects of miR-323b-5p suppression on LPS-induced injury in LPS-stimulated FHCs. Furthermore, the effects of ANRIL on LPS-induced cell injury were achieved by TLR4/MyD88/NF-κB pathway. Our data indicate that suppression of ANRIL may inhibit the development of UC by regulating miR-323b-5p/TLR4/MyD88/NF-κB pathway. ANRIL/miR-323b-5p/TLR4/MyD88/NF-κB pathway may provide a new strategy for UC therapy.     

藁本内酯神经炎症抑制作用与PPARγ依赖的TLR4/NF-κB信号通路的相关性

藁本内酯(LIG)具有抑制神经炎症反应和神经保护作用,TLR4/NF-κB作为脑内神经炎症应答最重要的通路之一,与过氧化物酶体增殖受体γ(PPARγ)的相互作用与大脑神经炎症应答及炎症损伤有关。本研究为阐明TLR4/NF-κB信号通路和PPARγ在LIG的神经炎症抑制作用中所发挥的作用,通过雄性大鼠侧脑室注射脂多糖(LPS)造成大鼠神经炎模型研究,并在注射LPS前预先给予溶媒、LIG(10 mg/kg,20 mg/kg)、GW9662(PPARγ选择性拮抗剂),探讨LIG对于LPS诱导的大鼠急性神经炎症模型的保护作用及机制。结果表明LIG对于LPS诱导的促炎症因子(TNF-α,MCP-1)的产生、TLR4/NF-k B/p38 MAPK信号通路的活化均有抑制作用,且具有剂量依赖性,同时能增强PPARγ转录因子活性,同样具有剂量依赖性。LIG对于LPS诱导的大鼠神经炎症的上述作用均可被GW9662拮抗。这些结果表明LIG通过调节PPARγ依赖的TLR4/NF-κB信号通路对LPS诱导的神经炎症起到抑制作用。     

基于TLR4/MyD88/NF-κB信号通路探讨安肠汤联合艾灸治疗肝郁脾虚证腹泻型肠易激综合征的疗效及其机制

摘要 目的：基于Toll样受体4（TLR4）/髓样分化因子88（MyD88）/核因子-κB（NF-κB）信号通路探讨安肠汤联合艾灸治疗肝郁脾虚证腹泻型肠易激综合征（IBS）的疗效及其机制。方法：采用随机数字表法,将广州中医药大学第一附属医院在2019年4月~2022年12月期间收治的108例腹泻型IBS患者分为对照组（常规药物联合艾灸治疗,n=54）和研究组（对照组基础上接受安肠汤治疗,n=54）。对比两组疗效、中医证候总积分、IBS症状严重程度问卷（IBS-SSS）评分、肠屏障功能指标、TLR4/MyD88/NF-κB信号通路相关信使核糖核酸（mRNA）表达水平。结果：研究组的临床总有效率高于对照组（P<0.05）。两组治疗后中医证候总积分、IBS-SSS评分下降,且研究组低于对照组（P<0.05）。两组治疗后肠脂肪酸结合蛋白（IFABP）、D-乳酸及二胺氧化酶（DAO）下降,且研究组低于对照组（P<0.05）。两组治疗后TLR4、MyD88、NF-κB mRNA表达下降,且研究组低于对照组（P<0.05）。结论：安肠汤联合艾灸治疗肝郁脾虚证腹泻型IBS患者,可有效改善临床症状和肠屏障功能,疗效较好,可能与调节TLR4/MyD88/NF-κB信号通路有关。     

有氧运动加螺旋藻补充对2型糖尿病大鼠肾脏TLR4/NF-κB-p65的表达及炎性因子的影响

本文旨在研究有氧运动加螺旋藻补充对2型糖尿病大鼠肾脏TLR4/NF-κB-p65的表达及炎性因子的影响,以TLR4/NF-κB-p65炎症信号通路为靶点探讨运动加螺旋藻补充改善2型糖尿病大鼠肾脏损伤的可能机制。采用4周高脂饲料喂养和低剂量STZ腹腔注射的方法建立2型糖尿病大鼠实验模型,成模后将大鼠随机分为糖尿病安静对照组(DM)、糖尿病运动组(DE)、糖尿病+螺旋藻组(DS)、糖尿病运动+螺旋藻组(DES),另设正常安静对照组(NC)。运动方式采用8周的无负重游泳训练。实验末,测随机血糖、微量白蛋白、肾脏测TNF-α、TLR4和NF-κB-p65蛋白表达,并采用光镜观察肾脏微细结构的改变。结果表明:(1)光镜下可见DE组、DS组和DES组大鼠肾小球毛细血管袢面积增加;肾小管扩张、上皮细胞空洞变性、细胞聚集增加和炎症细胞稍浸润等病理变化均较DM组有所改善。(2)DE组、DS组和DES组大鼠的血糖浓度、尿微量白蛋白含量均较DM组降低(P0.05或P0.01);与DES组比较,DE组和DS组的血糖浓度、尿微量白蛋白含量均升高明显(P0.05或P0.01)。(3)DE组、DS组和DES组大鼠肾脏TNF-α、TLR4和NF-κB-p65蛋白表达均明显降低,与DM组比较,差异均呈显著性(P0.01);与DES组比较,DE组和DS组的肾脏TNF-α、TLR4和NF-κB-p65蛋白表达明显增加(P0.05或P0.01)。因此,有氧运动、螺旋藻及两者联合使用有降低2型糖尿病大鼠血糖,改善肾功能,减轻肾脏损害的功效,但两者联合使用效果要好于单独使用,其机制可能与有氧运动联合螺旋藻降低TLR4/NF-κB-p65信号蛋白表达发挥抗炎作用的效果要好于单纯的有氧运动或螺旋藻使用。     

Avicularin alleviates acute liver failure by regulation of the TLR4/MyD88/NF-κB and Nrf2/HO-1/GPX4 pathways to reduce inflammation and ferroptosis

Acute liver failure (ALF) is an inflammation-mediated hepatocyte death process associated with ferroptosis. Avicularin (AL), a Chinese herbal medicine, exerts anti-inflammatory and antioxidative effects. However, the protective effect of AL and the mechanism on ALF have not been reported. Our in vivo results suggest that AL significantly alleviated lipopolysaccharide (LPS)/D-galactosamine (D-GalN)-induced hepatic pathological injury, liver enzymes, inflammatory cytokines, reactive oxygen species and iron levels and increased the antioxidant enzyme activities (malondialdehyde and glutathione). Our further in vitro experiments demonstrated that AL suppressed inflammatory response in LPS-stimulated RAW 264.7 cells via blocking the toll-like receptor 4 (TLR4)/myeloid differentiation protein-88 (MyD88)/nuclear factor kappa B (NF-κB) pathway. Moreover, AL attenuated ferroptosis in D-GalN-induced HepG2 cells by activating the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1)/glutathione peroxidase 4 (GPX4) pathway. Therefore, AL can alleviate inflammatory response and ferroptosis in LPS/D-GalN-induced ALF, and its protective effects are associated with blocking TLR4/MyD88/NF-κB pathway and activating Nrf2/HO-1/GPX4 pathway. Moreover, AL is a promising therapeutic option for ALF and should be clinically explored.     

乳源性复合益生菌对糖尿病大鼠肾组织 TLR2、TLR4/NFκB信号通路的影响

目的 研究乳源性复合益生菌对糖尿病大鼠肾组织Toll样受体2(TLR2)、Toll样受体4(TLR4)、核转录因子κB(NFκB)表达的影响。 方法 将高脂饮食和链脲佐菌素(STZ)诱导的糖尿病SD大鼠模型随机分为模型组、二甲双胍组、利拉鲁肽组、复合益生菌低剂量组和复合益生菌高剂量组,正常SD大鼠为对照组,每组8只。血糖仪检测不同时段血糖值,ELISA法检测糖化血红蛋白(HbA1c)含量,生化仪检测大鼠尿素氮(BUN)、肌酐(Scr)、24 h尿蛋白定量(24h Alb)的变化,HE染色观察肾脏组织形态,qPCR法检测肾组织TLR2、TLR4的mRNA的表达以及Western blot检测TLR2、NFκBpp65蛋白表达量。 结果 与模型组相比,复合益生菌组大鼠HbA1c、BUN、Scr及24h Alb水平明显下降,并且复合益生菌能够明显改善肾脏的组织形态,显著降低TLR2、TLR4的mRNA的表达和TLR2、NFκBpp65蛋白表达量。 结论 复合益生菌可显著改善糖尿病大鼠早期肾脏损害,其机制与部分抑制糖尿病大鼠肾组织中TLR2、TLR4/NFκB信号通路有关。