姜黄素对亚急性肝内胆汁淤积大鼠肠内菌群变化的影响及其机制研究

目的： 研究姜黄素对亚急性肝内胆汁淤积（intrahepatic cholestasis,IHC）大鼠肠内菌群变化的影响,并分析相关机制。方法： 38只IHC大鼠随机分为模型组（9只）、姜黄素低剂量组（9只）、姜黄素高剂量组（10只）、阳性药物组（10只）,9只健康大鼠设置为对照组。姜黄素低、高剂量组姜黄素200、400 mg·kg^-1灌胃;阳性药物组熊去氧胆酸60 mg·kg^-1灌胃;对照组与疾病组等量生理盐水灌胃。均为每天一次;干预14 d。检测肝功能指标、胆红素代谢指标、炎症指标;qRT-PCR反应定量检测肠内菌群;Western blot检测肝组织核转录因子-κB（NF-κB）p65、p-NF-κB p65、核因子抑制蛋白（IκBα）、p-IκBα蛋白相对表达量。结果： 与疾病组比较,姜黄素低、高剂量组、阳性药物组碱性磷酸酶（ALP）、丙氨酸氨基转移酶（ALT）、谷草转氨酶（AST）、γ-谷氨酰转肽酶（GGT）、总胆红素（TBIL）、直接胆红素（DBIL）、总胆汁酸（TBA）、白介素-1β（IL-1β）、白介素-6（IL-6）、转化生长因子-β₁（TGF-β₁）均降低,白介素-10（IL-10）均升高,乳酸杆菌、双歧杆菌均增加,大肠杆菌均减少（P<0.05）;与姜黄素低剂量组比较,姜黄素高剂量组、阳性药物组ALP、ALT、AST、GGT、TBIL、DBIL、TBA、IL-1β、IL-6、TGF-β₁均降低,IL-10均升高,乳酸杆菌、双歧杆菌均增加,大肠杆菌均减少（P<0.05）。与疾病组比较,姜黄素低、高剂量组、阳性药物组p-NF-κB p65/NF-κB p65,p-IκBα/IκBα均降低（P<0.05）;与姜黄素低剂量组比较,姜黄素高剂量组、阳性药物组p-NF-κB p65/NF-κB p65,p-IκBα/IκBα均降低（P<0.05）。结论： 姜黄素可改善亚急性IHC模型大鼠肝功能、胆红素代谢、肠道菌群,减轻炎症反应及肝脏组织病理变化,且具有剂量依赖性,推测其作用机制与抑制NF-κB信号通路有关。     

狐臭柴茎挥发油体外抗炎活性及基于NF-κB和MAPKs信号通路作用机制的研究

目的:探究狐臭柴茎挥发油的体外抗炎活性及其作用机制。方法:通过水蒸气蒸馏法制备狐臭柴茎挥发油,采用GC-MS对其化学成分进行分析。通过Griess法和ELISA法测定挥发油对LPS诱导RAW264.7细胞上清液中一氧化氮(NO)、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)水平的影响;qRT-PCR检测iNOS、COX-2、TNF-α、IL-6、IL-1β mRNA的表达;Western blot测定挥发油对iNOS、COX-2、NF-κB和MAPKs信号通路蛋白的影响。结果:从挥发油中共鉴定出74种化学成分,其中主要的化学成分是茅术醇(13.50%)。挥发油显著抑制NO、TNF-α和IL-6的分泌(P＜0.01),同时降低了促炎因子(TNF-α、IL-6和IL-1β)和促炎酶(iNOS和COX-2)的mRNA表达水平(P＜0.01)。Western blot研究表明挥发油能下调NF-κB信号通路细胞核p65蛋白表达、p65和IκBα磷酸化(P＜0.05或P＜0.01)。此外,还降低了MAPKs信号通路p38、JNK和ERK蛋白的磷酸化(P＜0.01)。结论:狐臭柴茎挥发油对LPS诱导RAW264.7细胞炎症模型具有较好的抗炎效果,其内在的分子机制与下调NF-κB和MAPKs信号通路有关。     

黄芩汤对非酒精性脂肪肝炎大鼠的作用及机制研究

目的:研究黄芩汤对非酒精性脂肪肝炎(non-alcoholic steatohepatitis,NASH)的作用及机制。方法:将大鼠随机分为空白组、模型组、阳性对照组(多烯磷脂胆碱,8 mg·kg^-1)、黄芩汤低、中、高剂量组(5,10,20 g·kg^-1),采用高脂饮食饲喂12周造模。灌胃给药5周后,检测各组大鼠血脂、肝功能及炎性因子水平;HE染色观察肝脏组织病理变化;RT-PCR和Western Blot法检测肝组织Toll样受体2(TLR2)、髓样分化因子88(MyD88)、核因子κB p65(NF-κB p65)、白介素6(IL-6)、维甲酸相关核孤儿受体γt(RORγt)、IL-23和IL-17 mRNA和蛋白表达;免疫组化法检测肝组织中NF-κB p65表达。结果:与空白组比较,模型组大鼠血脂、肝功能和炎性因子水平明显异常(P<0.01)。肝组织出现脂肪变性及炎性浸润;肝组织中TLR2、MyD88、NF-κB p65、IL-6、RORγt、IL-23和IL-17 mRNA和蛋白表达明显升高(P<0.01);NF-κB p65阳性细胞表达明显提升(P<0.01)。与模型组比较,黄芩汤组大鼠血脂、肝功能和炎性因子水平得到明显恢复(P<0.05,P<0.01);肝组织脂肪变性和炎性浸润得到纠正;上述蛋白mRNA和蛋白表达均明显降低(P<0.05,P<0.01);NF-κB p65阳性细胞表达明显降低(P<0.01)。结论:黄芩汤可能通过抑制TLR2/MyD88/NF-κB信号通路和IL-23/IL-17轴治疗NASH。     

罗浮山百草油抗炎作用机制及活性组分筛选研究

目的：研究罗浮山百草油体内及体外抗炎作用的药效及作用机制并初步筛选其活性组分。方法：分别使用二甲苯致小鼠耳肿胀法及脂多糖（lipopolysaccharide,LPS）诱导小鼠单核巨噬细胞白血病细胞RAW264.7炎性模型检测罗浮山百草油的体内及体外抗炎作用,使用RT-PCR法及ELISA法检测细胞内炎性因子变化;通过脂多糖LPS诱导人急性单核细胞白血病细胞THP1炎性模型,使用Western blot法检测核因子-κB p65（nuclear factor κB p65,NF-κB p65）及p38丝裂原活化蛋白激酶（p38 mitogen-activated protein kinase,p38 MAPK）的磷酸化水平。将罗浮山百草油分为13个组分,使用LPS诱导RAW264.7细胞炎性模型初步筛选其活性组分。结果：罗浮山百草油可剂量依赖性地抑制二甲苯诱导的小鼠耳肿胀及LPS诱导RAW264.7细胞一氧化氮（nitric oxide,NO）的产生,抑制细胞内白细胞介素1β（interleukin-1β,IL-1β）、白细胞介素6（interleukin-6,IL-6）、肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）、诱导型一氧化氮合酶（inducible nitric oxide synthase,iNOS） mRNA和IL-1β、IL-6蛋白的表达;罗浮山百草油可抑制LPS诱导THP1细胞NF-κB p65及p38 MAPK的磷酸化。活性组分初步筛选发现,13个组分中,百草精（68种药材的茶油提取物）、丁香罗勒油、肉桂油以及八角茴香油具有较强的抗炎活性,是罗浮山百草油抗炎活性的主要贡献者。结论：本研究证实了罗浮山百草油抗炎活性的有效性,并筛选出了相应活性组分,可为临床应用及质量控制提供依据。     

黄芩苷对胶原诱导性关节炎大鼠抗炎作用研究

目的研究黄芩苷对胶原诱导性关节炎(CIA)大鼠的抗炎作用并探讨其对白细胞介素(IL)-1β、肿瘤坏死因子(TNF)-α和核因子(NF)-κB信号通路的影响,为其应用于类风湿关节炎临床治疗提供实验依据。方法 Wistar雌性大鼠制备CIA模型,将关节炎评分>2的CIA大鼠采用随机数字表法随机分为模型对照组(0.9%氯化钠注射液)、甲氨蝶呤组(1 mg/kg)、黄芩苷低、中、高剂量组(50、100、200 mg/kg),每组6只持续处理30 d。另外选取6只正常大鼠作为正常对照(0.9%氯化钠注射液)。检测大鼠关节炎症状,关节组织进行放射评估和组织病理检测。定量检测血清中TNF-α和IL-1β水平,蛋白质印迹法检测关节滑膜组织中NF-κB p65、磷酸化-NF-κB p65(Ser536)和乙酰化-NF-κB p65(Lys310)蛋白表达。结果黄芩苷能减轻CIA大鼠的关节肿胀,明显抑制关节滑膜炎症和关节软骨及骨破坏,明显降低血清中TNF-α、IL-1β含量,抑制关节滑膜组织中NF-κB p65、磷酸化-NF-κB p65(Ser536)和乙酰化-NF-κB p65(Lys310)蛋白水平表达。结论黄芩苷抑制CIA大鼠炎症可能是通过抑制炎性细胞因子的产生和NF-κB信号转导途径而发挥治疗作用的。     

中药排气汤对术后肠梗阻小鼠胃肠功能的作用及机制研究

目的:研究中药排气汤对术后肠梗阻小鼠胃肠功能的作用及机制。方法:取7周龄SPF级C57BL/6J小鼠24只,随机分为假手术组、模型组、莫沙必利组、排气汤组,每组6只,采用经典小肠干扰术诱导小鼠术后肠梗阻模型。手术24 h后观测各组小鼠胃肠运动功能、肠道组织结构,ELISA试剂盒检测小鼠血清促炎因子白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)和抗炎因子白细胞介素-10(IL-10)的水平,免疫组织化学法检测小鼠小肠Cajal间质细胞(interstitial cells of Cajal,ICC)及诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)的表达情况,Western Blot检测小鼠小肠中Toll样受体4(TLR4)、活化丝裂原激活的蛋白激酶p38(p38MAPK)和核转录因子-κB p65(NF-κB p65)表达水平。结果:与假手术组相比,模型组小鼠小肠推进率显著下降(P<0.01),血清炎症因子含量明显升高(P<0.05);与模型组相比,排气汤组小鼠小肠推进率明显升高(P<0.05),血清中炎症因子含量明显降低(P<0.05)。Western Blot结果显示,排气汤组能显著降低术后肠梗阻小鼠小肠TLR4、p38MAPK和NF-κB p65蛋白表达(P<0.05)。结论:中药排气汤可以改善术后肠梗阻小鼠胃肠道功能,其作用机制与恢复ICC的功能,抑制TLR4、p38MAPK和NF-κB p65的表达,减轻炎症反应有关。     

黄芪多糖抑制NF-κB/MAPK信号通路和改善哮喘大鼠气道炎症的作用

目的观察黄芪多糖(astragalus polysaccharide,APS)对哮喘大鼠气道炎症及NF-κB/MAPK信号通路的影响。方法采用卵蛋白(ovalbumin,OVA)致敏法制备哮喘模型并予以药物治疗。观察支气管灌洗液(bronchoalveolar lavage fluid,BALF)中炎症细胞分类计数、肺组织病理变化、肺泡Ⅰ型上皮超微结构变化;测定肺组织NF-κB p65、磷酸化NF-κB p65(p-NF-κB p65)、磷酸化IκBα(p-IκBα)、ERK1/2、磷酸化ERK1/2(p-ERK1/2)、JNK、磷酸化JNK(p-JNK)、p38 MAPK、磷酸化p38 MAPK(p-p38 MAPK)、IL-1β、IL-4、IL-5、IL-6及IL-13含量的变化。结果与正常组比较,OVA致敏明显升高哮喘组炎症细胞数量、增强NF-κB/MAPK信号通路活性。APS可改善哮喘大鼠气道炎症、肺Ⅰ型上皮损伤,并明显抑制NF-κB/MAPK信号通路活性。结论 APS改善哮喘大鼠气道炎症、细胞损伤的机制可能与抑制NF-κB/MAPK信号通路有关。     

双氢青蒿素通过抑制炎症反应在血管重塑中的作用机制研究

目的 探究双氢青蒿素通过抑制炎症反应对血管重塑的抑制作用。方法 采用贴块法培养小鼠血管平滑肌细胞(vascular smooth muscle cells，VSMCs)，以TNF-α诱导建立体外平滑肌细胞增殖模型，通过手术剥夺动脉方式建立小鼠股动脉损伤模型，并采用双氢青蒿素干预细胞及股动脉损伤小鼠。采用CCK-8法检测细胞增殖；H&E染色评价股动脉组织病理变化情况并测量中膜厚度；ELISA法检测细胞中IL-1β、TNF-α、IL-6、CCR8及小鼠股动脉组织中IL-1β、TNF-α、IL-6、TGF-β1的含量；免疫组织化学法检测股动脉组织NF-κB p65、CCR8表达；qRT-PCR分析细胞及股动脉组织中IL-1β、TNF-α、IL-6、IL-8、CCR8、NF-κB p65、TGF-β1、MCP-1 mRNA表达；Western blotting分析细胞及股动脉组织中IL-1β、IL-6、TNF-α、IL-8、p-NF-κB p65、NF-κB p65、CCR8、TGF-β1、MCP-1蛋白表达。结果 在TNF-α诱导的VSMCs增殖及股动脉损伤小鼠模型中，VSMCs增殖及TNF-α、IL-6含量明显升高，IL-1β、IL-6、IL-8 mRNA表达明显升高，IL-6、CCR8、NF-κB p65蛋白表达明显升高(P<0.05或P<0.01)；双氢青蒿素可明显降低TNF-α诱导的VSMCs中IL-1β、IL-8 mRNA及股动脉损伤小鼠IL-1β mRNA表达和IL-1β、p-NF-κB p65蛋白表达(P<0.05或P<0.01)。结论 双氢青蒿素能降低血管炎症，延缓损伤血管的重塑，表明其为经皮冠状动脉介入治疗(如支架植入)的潜在治疗药物。     

鼠曲草提取物对COPD模型大鼠气道炎症的抑制作用及对NF-κB通路的影响

目的:研究鼠曲草提取物对慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)模型大鼠气道炎症的影响及作用机制。方法:大鼠随机分为正常组,模型组,鼠曲草提取物低、中、高剂量组(0.75,1.5,3g·kg^-1)和地塞米松组(阳性对照药,27mg·kg^-1),每组9只。除正常组外,其余各组大鼠采用香烟烟熏联合气管内滴注脂多糖建立COPD大鼠模型。造模15d后,各给药组大鼠灌胃相应药物,正常组和模型组大鼠灌胃等体积0.9%氯化钠溶液,每日1次,连续14d。末次给药后,检测各组大鼠肺功能;HE染色观察肺组织病理学变化;酶联免疫法检测肺泡灌洗液、血清中IL-6、IL-8、IL-17、IL-1β、TNF-α、TGF-β含量;RT-PCR法检测右肺组织中NF-κB mRNA的相对表达量;Western blot法检测右肺组织中NF-κB、IKKβ、IκBα及其p-IκBα蛋白的相对表达量。结果:不同剂量的鼠曲草提取物干预后,均可显著提高大鼠肺功能参数FEV0.3、FVC、FEV0.3/FVC,明显降低肺泡灌洗液中IL-8、IL-17、TNF-α及血清中IL-1β、IL-6、IL-8、IL-17、TNF-α、TGF-β含量,改善大鼠肺组织病理变化,同时显著降低p-IκB、IKK蛋白及NF-κB mRNA和蛋白相对表达量。结论:鼠曲草能够改善COPD模型大鼠的肺功能,减轻肺组织病理损伤,抑制炎症反应,其作用可能与抑制NF-κB信号通路有关。     

丹皮酚对脂多糖诱导的星形胶质细胞炎性因子分泌的影响

目的观察丹皮酚对体外培养的星形胶质细胞炎性因子分泌的影响,并探讨其作用机制。方法采用神经胶质原纤维酸性蛋白(GFAP)免疫荧光染色法鉴定星形胶质细胞;实验分为对照组,模型组和2.5、5、10μmol·L-1丹皮酚组,0.5 mg·L-1脂多糖(LPS)诱导炎症反应。采用ELISA法测定培养液中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)水平;采用Western blot检测细胞IκBα蛋白表达和磷酸化水平及胞核NF-κB(p65)蛋白表达水平。结果与对照组相比,模型组星形胶质细胞上清液中IL-1β、IL-6和TNF-α水平显著增加(P<0.01),胞浆IκBα蛋白表达受抑(P<0.01),IκBα蛋白磷酸化和胞核NF-κB(p65)蛋白表达水平上调(P<0.01);5、10μmol·L-1丹皮酚能减少LPS活化的星形胶质细胞上清液中IL-1β、IL-6和TNF-α水平(P<0.05或P<0.01),增加胞浆IκBα蛋白表达(P<0.05或P<0.01),抑制LPS上调的IκBα蛋白磷酸化和胞核NF-κB(p65)蛋白表达水平(P<0.05或P<0.01)。结论丹皮酚能抑制LPS诱导的星形胶质细胞炎性因子IL-1β、IL-6和TNF-α的分泌,IκBα/NF-κB信号通路可能参与了丹皮酚对星形胶质细胞炎症反应的抑制作用。     

Dioscin reduces lipopolysaccharide-induced inflammatory liver injury via regulating TLR4/MyD88 signal pathway

We previously reported the effects of dioscin against carbon tetrachloride-, acetaminophen- and alcohol-induced acute liver damage. However, its effect on lipopolysaccharide (LPS)-induced inflammatory liver injury remains unknown. In the present work, liver injury in mice and rats was induced by LPS, and dioscin was intragastrically administered for 7 days. In vitro, the AML-12 cells and HepG-2 cells were treated with LPS after dioscin treatment. The results showed that dioscin not only markedly reduced serum ALT, AST levels and relative liver weights, but also restored cell injury caused by LPS. In mechanism study, dioscin significantly attenuated inflammation through down-regulating the levels of toll-like receptor (TLR) 4, myeloid differentiation factor 88 (MyD88), interleukin-1 receptor-associated kinase 1 (IRAK1), tumor necrosis factor receptor-associated factor 6 (TRAF6), phosphorylated inhibitor of nuclear factor κB kinase (p-IKK), phosphorylated inhibitor of nuclear factor κB alpha (p-IκBα), phosphorylated nuclear factor κB p65 (p-NF-κB p65), high-mobility group protein 1 (HMGB-1), interleukin (IL)-1, IL-6 and tumor necrosis factor-α (TNF-α). TLR4 overexpression was also decreased by dioscin, leading to the markedly decreased levels of MyD88, IRAK1, TRAF6, p-IKK, p-IκBα, p-NF-κB p65 and HMGB-1. Suppression of MyD88 by ST2825 eliminated the inhibitory effects of dioscin on the levels of IRAK1, TRAF6, p-IKK, p-IκBα, p-NF-κB p65, HMGB-1, IL-1β, IL-6 and TNF-α. Our results suggested that dioscin exhibited protective effect against LPS-induced liver injury via altering TLR4/MyD88 pathway, which should be developed as one potent candidate for the treatment of acute inflammatory liver injury in the future.     

Anti-inflammatory effect of Yam Glycoprotein on lipopolysaccharide-induced acute lung injury via the NLRP3 and NF-κB/TLR4 signaling pathway

Acute lung injury (ALI) is a common lung disease accompanied by acute and persistent pulmonary inflammatory response syndrome, which leads to alveolar epithelial cells and capillary endothelial cell damage. Yam glycoprotein, separated from traditional Chinese yam, has been shown to have anti-inflammatory and immunomodulatory effects. In this experiment, we mainly studied the therapeutic effect and mechanism of a glycoprotein on the lipopolysaccharide (LPS)-induced ALI mice. An oral glycoprotein method was used to treat the mouse ALI model induced by LPS injection in the peritoneal cavity. Afterward, we measured the wet/dry (W/D) ratio, the activity of myeloperoxidase (MPO), the oxidative index superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GSH-PX) and the production of inflammatory cytokines interleukin-1β (IL-1β), tumour necrosis factor-α (TNF-α), and interleukin-6 (IL-6) to evaluate the effect of yam glycoprotein on lung tissue changes. We examined the protein expression of TLR4, ASC, NF-κBp65, p-NF-κBp65, Caspase-1, IκB, NLRP3, p-IκB, and β-actin by western blot analysis. Immunohistochemical analyses of NLRP3 and p-p65 in lung tissue were carried out to assess the mechanism of glycoprotein action. This result suggests that glycoprotein markedly depressed LPS-induced lung W/D ratio, MPO activity, MDA content SOD and GSH-Px depletion, and the contents of inflammatory cytokines IL-1β, IL-6, and TNF-α. Moreover, glycoprotein blocked TLR4/NF-κBp65 signaling activation and NLRP3inflammasome expression in LPS-induced ALI mice. As this particular study shows, glycoprotein has a safeguarding effects on LPS-induced ALI mice, possibly via activating NLRP3inflammasome and TLR4/NF-κB signaling pathways.     

Tetrachlorobenzoquinone exhibits neurotoxicity by inducing inflammatory responses through ROS-mediated IKK/IκB/NF-κB signaling

Tetrachlorobenzoquinone (TCBQ) is a joint metabolite of persistent organic pollutants (POPs), hexachlorobenzene (HCB) and pentachlorophenol (PCP). Previous studies have been reported that TCBQ contributes to acute hepatic damage due to its pro-oxidative nature. In the current study, TCBQ showed the highest capacity on the cytotoxicity, ROS formation and inflammatory cytokines release among four compounds, i.e., HCB, PCP, tetrachlorohydroquinone (TCHQ, reduced form of TCBQ) and TCBQ, in PC 12 cells. Further mechanistic study illustrated TCBQ activates nuclear factor-kappa B (NF-κB) signaling. The activation of NF-κB was identified by measuring the protein expressions of inhibitor of nuclear factor kappa-B kinase (IKK) α/β, p-IKKα/β, an inhibitor of NF-κB (IκB) α, p-IκBα, NF-κB (p65) and p-p65. The translocation of NF-κB was assessed by Western blotting of p65 in nuclear/cytosolic fractions, electrophoretic mobility shift assay (EMSA) and luciferase reporter gene assay. In addition, TCBQ significantly induced protein and mRNA expressions of inflammatory cytokines and mediators, such as interleukin-1 beta (IL-1β), IL-6, tumor necrosis factor-alpha (TNF-α), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and the production of nitric oxide (NO) and prostaglandin E2 (PGE2). Pyrrolidine dithiocarbamate (PDTC), a specific NF-κB inhibitor inhibited these effects efficiently, further suggested TCBQ-induced inflammatory responses involve NF-κB signaling. Moreover, antioxidants, i.e., N-acetyl-l-cysteine (NAC), Vitamin E and curcumin, ameliorated TCBQ-induced ROS generation as well as the activation of NF-κB, which implied that ROS serve as the upstream molecule of NF-κB signaling. In summary, TCBQ exhibits a neurotoxic effect by inducing oxidative stress-mediated inflammatory responses via the activation of IKK/IκB/NF-κB pathway in PC12 cells.     

五灵胶囊中部分化学成分对脂多糖诱导枯否细胞释放炎症因子的影响

目的观察五灵胶囊（Wuling capsules,WL）中部分化学成分对脂多糖（lipopolysaccharider,LPS）诱导大鼠原代枯否细胞（Kupffer cell,KC）表达ERK（extracellular signal-regulated kinase）和核因子（nuclear factor-kappa B,NF-κB）调节炎症因子和介质的作用。方法分离大鼠KC,采用60μg L 1 LPS诱导KC分泌炎症因子及NOS,ELISA测定TNF-α、IL-6、IL-8,比色法测定NOS,Western blot检测全蛋白ERK、p-ERK、NF-κBp50、NF-κBp65、p-NF-κBp65、p-IκK、p-IκB及核、浆蛋白NF-κBp65、p-NF-κBp65的变化。结果 WL明显下调LPS活化KC表达p-ERK、p-NF-κBp65、p-IκK、p-IκB蛋白,模拟WL混合成分（Mix）及6单体成分明显下调LPS活化KC表达p-ERK和p-NF-κBp65信号通路蛋白。各受试药物组均能降低核内p-NF-κBp65表达,减少p-NF-κBp65入核抑制炎性基因转录,降低活化KC过量分泌TNF-α、IL-6、IL-8和NOS。结论 WL中五味子醇甲、五味子乙素、隐丹参酮、丹参酮ⅡA、柴胡皂苷D是干预LPS诱导KC表达ERK、NF-κB信号通路蛋白,抑制炎性因子与介质基因转录而减少TNF-α、IL-6、IL-8和NOS分泌的有效成分。     

Cannabinoid receptor agonist WIN55,212-2 and fatty acid amide hydrolase inhibitor URB597 ameliorate neuroinflammatory responses in chronic cerebral hypoperfusion model by blocking NF-κB pathways

The present study explored the protective effects of cannabinoid receptor agonist WIN55,212-2 (WIN) and fatty acid amide hydrolase inhibitor URB597 (URB) against neuroinflammation in rats with chronic cerebral hypoperfusion (CCH). Activated microglia, astrocytes, and nuclear factor kappa B (NF-κB) p65-positive cells were measured by immunofluorescence. Reactive oxygen species (ROS) was assessed by dihydroethidium staining. The protein levels of cluster of differentiation molecule 11b (OX-42), glial fibrillary acidic protein (GFAP), NF-κB p65, inhibitor of kappa B alpha (IκB-a), IκB kinase a/β (IKK a/β), phosphorylated IKK a/β (p-IKK a/β), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF)-α, and interleukin-1β (IL-1β) were examined by western blotting or enzyme-linked immunosorbent assay. All the protein levels of OX-42, GFAP, TNF-a, IL-1β, COX-2, and iNOS are increased in CCH rats. WIN and URB downregulated the levels of OX-42, GFAP, TNF-α, IL-1β, COX-2 and iNOS and inhibited CCH-induced ROS accumulation in CCH rats, indicating that WIN and URB might exert their neuroprotective effects by inhibiting the neuroinflammatory response. In addition, the NF-κB signaling pathway was activated by CCH in frontal cortex and hippocampus, while the aforementioned changes were reversed by WIN and URB treatment. These findings suggest that WIN and URB treatment ameliorated CCH-induced neuroinflammation through inhibition of the classical pathway of NF-κB activation, resulting in mitigation of chronic ischemic injury.

     

柴芩承气汤通过抑制TLR4/NF-κB p65通路减轻小鼠重症急性胰腺炎并发肝损伤

目的基于TLR4/NF-κB p65信号通路探讨柴芩承气汤(chaiqinchengqi decoction,CQCQD)对小鼠重症急性胰腺炎(severe acute pancreatitis,SAP)并发肝损伤的保护机制。方法昆明小鼠36只,随机分为3组(n=12),即对照组(Control),重症急性胰腺炎模型组(SAP)和柴芩承气汤治疗组(SAP+CQCQD)。腹腔注射20%L-精氨酸(3.3 g·kg-1,2次,间隔1 h)建立SAP模型,治疗组给予柴芩承气汤灌胃(19 g·kg-1·d-1)。造模后72 h观察胰腺、肝脏组织病理变化,检测血清内毒素含量,肝组织TLR4、p-NF-κB p65蛋白表达,及肝内炎性因子水平。结果与Control组相比,SAP组胰腺和肝脏可见明显的病理损伤,血清内毒素含量增多,肝组织TLR4、p-NF-κB p65表达增加,IL-6、TNF-α、MIP-1αmRNA水平升高。与SAP组相比,柴芩承气汤组胰腺和肝脏组织病理损伤减轻,血清内毒素含量降低,肝组织TLR4、p-NF-κB p65表达和IL-6、TNF-α、MIP-1α mRNA水平减少。 结论 柴芩承气汤可能通过抑制肝组织TLR4/NF-κB p65通路活化,降低促炎因子水平,从而减轻小鼠SAP并发肝损伤。