姜黄素对TNF-α诱导人肾小球系膜细胞增殖及PTX3表达的作用研究

目的本实验研究姜黄素(Cur)对TNF-α诱导人肾小球系膜细胞过度增殖以及PTX3过度表达的影响,并对其作用机制进行初步探究。方法本实验选择人肾小球系膜细胞进行体外研究。实验共分为6组:空白对照组、TNF-α组、Cur 20,40和80μmol·L~(-1)组和PDTC组。1. Cur对TNF-α诱导人肾小球系膜细胞过度增殖及PTX3过度表达的影响。(1)使用MTT法检测Cur对人肾小球系膜细胞过度增殖的影响。(2)使用免疫荧光法检测Cu对人肾小球系膜细胞PTX3过度表达的影响。2. Cur对TNF-α诱导人肾小球系膜细胞PTX3过度表达的作用机制研究。(1)使用Western印迹法检测Cur对人肾小球系膜细胞内NF-κB信号通路中蛋白水平及PTX3蛋白表达的影响。(2)使用qRT-PCR法检测Cu对人肾小球系膜细胞内NF-κB信号通路中mRNA水平及PTX3 mRNA表达的影响。结果 (1)Cur对TNF-α诱导人肾小球系膜细胞PTX3的过度表达有良好的抑制作用。(2) Western印迹结果表明,Cur明显抑制NF-κB P65和IKKβ的蛋白表达,增加IκBα的蛋白表达。(3)qRT-PCR结果表明,Cur明显抑制NF-κB、P65和IKKβm RNA表达,增加IκBα的mRNA表达。结论 (1) Cur可有效抑制TNF-α诱导人肾小球系膜细胞的过度增殖。(2) Cur对TNF-α诱导人肾小球系膜细胞PTX3的过量表达产生良好的抑制效果,极有可能是通过抑制NF-κB信号通路的激活而达到。     

基于TLR4/NF-κB/NLRP3信号通路研究三叶香茶菜含药血清对枯否细胞活化的影响

目的 研究三叶香茶菜含药血清(isodon ternifolius-containing serum，ITS)通过Toll样受体4(TLR4)/核因子κB(NF-κB)/NOD样受体蛋白3(NLRP3)信号通路对脂多糖(lipopolysaccharide，LPS)诱导的大鼠原代肝枯否细胞(Kupffer cell，KC)活化的影响。方法 分离培养大鼠原代KC，将LPS诱导的大鼠原代KC分为空白对照组、模型对照组、空白血清组、阳性对照组(秋水仙碱含药血清组)、ITS组、TLR4阻断剂组、TLR4阻断剂+ITS组。MTT法检测不同浓度ITS对KC增殖活性的影响；ELISA法检测KC细胞上清液白介素-1β(IL-1β)、白细胞介素-18(IL-18)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)含量；荧光定量聚合应(PCR)、Western blotting和免疫荧光检测KC中TLR4/NF-κB/NLRP3信号通路中TLR4、核因子κB抑制蛋白α(IκBα)、半胱肽氨酸蛋白酶-1(Caspase-1)、NLRP3 mRNA和TLR4、IκBα、磷酸化IκBα(p-IκBα)、Caspase-1、NLRP3、NF-κBp65蛋白表达情况。结果 与模型对照组相比，各药物组KC上清液中IL-1β、IL-18、TNF-α和IL-6含量，以及细胞中TLR4、IκBα、Caspase-1、NLRP3 mRNA和TLR4、IκBα、p-IκBα、Caspase-1、NLRP3、NF-κBp65蛋白的表达均下调或降低(P<0.05或P<0.01)；与TLR4阻断剂组比较，TLR4阻断剂+ITS组上述多数指标的改善更加明显。结论 三叶香茶菜可能通过下调TLR4/NF-κB/NLRP3信号通路抑制KC活化，减少炎性因子的表达和释放，从而减轻肝脏炎症损伤。     

金骨莲胶囊对炎症模型大鼠的抗炎作用及机制研究

目的:研究金骨莲胶囊对炎症模型大鼠的抗炎作用及机制。方法:将48只大鼠随机分为空白对照组、模型组、金骨莲胶囊低、中、高剂量组(0.66、1.32、2.64 g/kg)和地塞米松组(阳性对照,0.945 mg/kg),每组8只。空白对照组和模型组大鼠灌胃等体积水,其余各组大鼠灌胃相应药物,每天给药2次,连续3天。末次给药后30 min,模型组和给药组大鼠腹腔注射脂多糖(10 mg/kg)以复制炎症模型。腹腔注射6 h后,于大鼠尾静脉取血,采用酶联免疫吸附试验检测大鼠血清中肿瘤坏死因子(TNF-α)、白细胞介素1β(IL-1β)、IL-6、前列腺素E(2 PGE2)的含量;测定大鼠肺组织湿质量/干质量(W/D)比值;采用苏木精-伊红染色法观察大鼠肺组织的病理学形态变化;采用逆转录-聚合酶链式反应法检测大鼠肺组织中TNF-α、IL-6、PGE2、IL-1βmRNA的表达水平;采用Western blot法检测大鼠肺组织中核因子κB(NF-κB)p65蛋白的磷酸化水平和NF-κB抑制蛋白α(IκBα)蛋白的表达水平。结果:与空白对照组比较,模型组大鼠血清中TNF-α、IL-1β、IL-6、PGE2的含量,肺组织W/D比值,肺组织中TNF-α、IL-1β、IL-6、PGE2 mRNA的表达水平和NF-κB p65蛋白磷酸化水平均显著升高,IκBα蛋白表达水平显著降低(P<0.05或P<0.01);大鼠肺组织中大量肺泡萎缩塌陷、壁增厚,可见肺实质化及大量炎症细胞浸润。与模型组比较,金骨莲胶囊各剂量组大鼠血清中TNF-α、IL-1β(低剂量组除外)、IL-6、PGE2的含量,以及肺组织中TNF-α(低剂量组除外)、IL-1β、IL-6(低、高剂量组除外)、PGE2 mRNA的表达水平均显著降低(P<0.05或P<0.01);金骨莲胶囊高剂量组大鼠肺组织W/D比值显著降低(P<0.05或P<0.01);金骨莲胶囊中剂量组大鼠肺组织中NF-κB p65蛋白磷酸化水平均显著降低(P<0.05或P<0.01),IκBα蛋白表达水平显著升高(P<0.05);大鼠肺泡结构较为清晰、壁轻度增厚,并见少量炎症细胞浸润。结论:金骨莲胶囊对炎症模型大鼠具有良好的抗炎作用,其作用机制可能与抑制NF-κB信号通路有关。     

生松素对AngⅡ诱导的大鼠肾小球系膜细胞细胞外基质合成的影响及其机制研究

目的 探讨生松素（Pb）对血管紧张素Ⅱ（AngⅡ）诱导的大鼠肾小球系膜细胞细胞外基质（ECM）合成的影响及其机制。方法 以大鼠肾小球系膜细胞为研究对象,将实验分为以下5组：正常对照（NC）组、1×10-6mol/L AngⅡ（Ng）组、1×10-6 mol/L AngⅡ+1‰ DMSO（Ng-D）组、 1×10-6 mol/L AngⅡ+30 μmol/L Pb（Ng-Pb）组、1×10-6 mol/L AngⅡ+10 μmol/L缬沙坦（Ng-Val）组。分别在干预12、24、48 h搜集细胞和上清液,采用Western blot检测Ⅳ型胶原（ColⅣ）、纤连蛋白（FN）、转化生长因子-β（ 1 TGF-β1 ）、Smad3、磷酸化Smad3（p-Smad3）、核因子κB（NF-κB）、磷酸化NF-κB（pNF-κB）的蛋白水平,酶联免疫吸附法（ELISA）检测上清液中肿瘤坏死因子α（TNF-α）、白细胞介素6（IL-6）水平,实时荧光定量PCR（qPCR）检测细胞中ColⅣ、FN、TGF-β1、TNF-α、IL-6的mRNA水平。结果 （1）Pb对 AngⅡ诱导的系膜细胞合成ECM的影响：与NC组相比,Ng组细胞内ColⅣ和FN的蛋白质和mRNA 水平升高（P＜0.05）,而Ng-Pb组和Ng-Val组细胞内ColⅣ和FN的蛋白质和mRNA水平均较Ng组降低（P＜0.05）。（2）Pb对TGF-β1/Smad3通路的 影响：与NC组相比,Ng组细胞内TGF-β1的蛋白质和mRNA水平增加,p-Smad3/Smad3水平增加（ P＜0.05）,而Ng-Pb组和Ng-Val组细胞内TGF-β1的蛋白质和mRNA水平以及p-Smad3/Smad3水平均较Ng组降低（P＜0.05）。（3）Pb对NF-κb介导的促炎症因子产生的影响：与NC组相比,Ng组 p-NF-κb/NF-κb水平增加,IL-6和TNF-α的蛋白质和mRNA水平增加（P＜0.05）,而Ng-Pb组和 Ng-Val组p-NF-κb/NF-κb水平以及IL-6和TNF- α的蛋白质和mRNA水平均较Ng组降低（P＜0.05）。结论 Pb可能通过抑制TGF-β1/Smad3信号通路和NF-κb介导的促炎症因子的产生, 从而抑制AngⅡ诱导的大鼠系膜细胞合成ECM。     

己酮可可碱联合骨髓间充质干细胞对高糖诱导的肾小球系膜细胞炎症因子及NF-κB信号通路的影响

目的 探讨己酮可可碱（Pentoxifylline,PTX）是否促进骨髓间充质干细胞（Mesenchymal stem cells,MSCs）对高糖作用下肾小球系膜细胞炎症因子及NF-κB信号通路的影响。方法 体外高糖培养小鼠肾小球系膜细胞,分为6组：对照组;模型组组;MSCs组;MSCs+ PTX 0.1 mM组;MSCs+ PTX 0.3 mM组和MSCs+ PTX 1 mM组。采用ELISA检测各组系膜细胞IL-6和TNF-α含量,采用Western blot检测各组系膜细胞中NF-?B p65,IKKα及IKKβ蛋白的表达。结果 与对照组相比,模型组系膜细胞的IL-6和TNF-α含量显著增高（P<0.01）,NF-?B p65、IKKα及IKKβ蛋白相对表达量显著升高（P<0.05或P<0.01）。与模型组相比,MSCs组及不同浓度PTX干预的系膜细胞分泌的IL-6及TNF-α含量出现明显降低（P<0.01）,NF-?B p65、IKKα及IKKβ蛋白表达量显著降低（P<0.05或P<0.01）。与MSCs组比较,MSCs+PTX 0.1 mM组、MSCs+ PTX 0.3 mM组及MSCs+PTX 1 mM组的IL-6和TNF-α含量显著下降（P<0.05或P<0.01）,NF-?B p65、IKKα和IKKβ的蛋白相对表达量明显降低（P<0.05或P<0.01）,且呈现PTX浓度依赖性。结论 不同浓度的PTX可通过抑制NF-?B信号通路的激活以及介导炎症反应促进MSCs对DN的治疗作用,且呈浓度依赖性。     

黄芩总黄酮对慢性肾功能衰竭大鼠的肾保护作用及ROS/MAPK/NF-κB信号通路的影响

摘要：目的:探讨黄芩总黄酮对慢性肾功能衰竭大鼠的肾保护作用及活性氧/丝裂原活化蛋白激酶/核因子κB（ROS/MAPK/NF-κB）信号通路的影响。方法:100只SD大鼠随机分为5组:正常对照组、模型组、海昆肾喜组(150 mg·kg-1)、黄芩总黄酮低(100 mg·kg-1)、高(200 mg·kg-1)剂量组,每组20只。除正常对照组外,其余各组建立慢性肾功能衰竭模型。建模成功后灌胃给予相应药物,1次/d,连续4周,正常对照组和模型组给予等体积生理盐水。实验结束后,测定24 h尿蛋白、肾/体比值、肌酐（SCr）、尿素（BUN）、肾小球细胞凋亡水平、肾脏ROS水平、肾脏MAPK、NF-κB基因和蛋白水平。结果:与正常对照组比较,模型组大鼠BUN、SCr、24 h尿蛋白、肾/体比、肾小球凋亡率、肾小球ROS、肾小球MAPK、NF-κB mRNA和蛋白水平显著升高（P<0.05）。与模型组比较,海昆肾喜组、黄芩总黄酮低、高剂量组大鼠BUN、SCr、24 h尿蛋白、肾/体比、肾小球凋亡率、肾小球ROS、肾小球MAPK、NF-κB mRNA和蛋白水平显著降低（P<0.05）,且黄芩总黄酮高剂量组各指标显著低于低剂量组（P<0.05）;黄芩总黄酮低剂量组大鼠各指标显著高于海昆肾喜组（P<0.05）,黄芩总黄酮高剂量组与海昆肾喜组相比差异无统计学意义（P>0.05）。模型组小鼠基底膜、肾小球系膜细胞明显增生肿胀,伴炎症粒细胞浸润;海昆肾喜组及黄芩总黄酮高剂量组肾小球系膜细胞轻度增生,炎症细胞浸润减少;黄芩总黄酮低剂量组仍然可见明显炎症细胞浸润,肾小球肿胀增生。结论:黄芩总黄酮对慢性肾功能衰竭大鼠具有肾保护作用,能明显降低肾小球凋亡率,其机制可能与黄芩总黄酮抑制氧化应激炎症反应通路ROS/MAPK/NF-κB激活有关。     

Ghrelin通过核因子-κB抑制肿瘤坏死因子-α诱导的HepG2细胞PAI-1 mRNA表达

目的探讨Ghrelin对肿瘤坏死因子-α(TNF-α)诱导的HepG2细胞纤溶酶原激活物抑制剂-1(PAI-1)mRNA表达的影响及核因子-κB(NF-κB)在其中的作用.方法HepG2细胞培养,加入不同浓度TNF-α,采用逆转录-聚合酶链反应测定(RT-PCR)检测PAI-1 mRNA的水平;免疫印迹法检测胞浆胞核NF-κBp65和胞浆κB抑制蛋白(IκB)的表达.给予Ghrelin预处理1 h后,加入TNF-α检测NF-κKBp65和IκB表达的变化.结果TNF-α(0.1,1,10μg·L-1)浓度依赖地增高HepG2细胞PAI-1 mRNA的表达;Ghrelin组的PAI-1 mRNA表达减少.TNF-α组胞核NF-κBp65表达增加,胞浆IκBα的表达减少;Ghrelin组较TNF-α组胞核NF-κBp65减少,胞浆IκBα的表达增加.结论TNF-α通过NF-κB介导HepG2 PAl-1mRNA的表达;Ghrelin通过抑制NF-κB而抑制TNF-α诱导PAI-1mRNA的表达.     

DATS通过NF-κB抑制LPS诱导小鼠肺泡巨噬细胞促炎细胞因子表达

目的探讨二烯丙基三硫(DATS)抑制脂多糖(LPS)诱导小鼠肺泡巨噬细胞肿瘤坏死因子-α(TNF-α)及白介素-1β表达的信号转导机制。方法体外培养MH-S细胞,用DATS和(或)LPS进行干预。反转录PCR检测细胞中TNF-α、IL-1β mRNA表达,电泳迁移率改变分析(EMSA)检测细胞核因子-κB(NF-κB)活性,Western blot检测细胞磷酸化(p-IκB)及非磷酸化IκB的表达。结果LPS刺激MH-S细胞可导致TNF-α、IL-1β mRNA、p-IκB表达增加及NF-κB活性升高。用DATS(0.1、0.5、2.5、5.0mg.L-1)预处理细胞30min后再给予LPS刺激,可使TNF-α、IL-1β mRNA表达降低,并呈剂量依赖性;升高的NF-κB活性及p-IκB表达均显不同程度的抑制。单独DATS对TNF-α、IL-1β mRNA表达及NF-κB活性无影响。结论DATS可通过抑制IκB磷酸化及NF-κB活化,进而下调LPS诱导小鼠肺泡巨噬细胞TNF-α、IL-1β mRNA表达。     

抑制NF-κB信号通路对顺铂致肺癌大鼠肾损伤的保护作用及其分子机制研究

摘要：目的:探索抑制核转录因子（NF）-κB信号通路对顺铂致肺癌大鼠肾损伤的保护作用及其分子机制研究。方法:将50只大鼠随机分为5组:正常对照组、肺癌组（Lewis细胞）、顺铂组(Lewis细胞+腹腔注射顺铂溶液5 mg·kg-1)、吡咯烷二硫代氨基甲酸盐（PDTC）组(Lewis细胞+腹腔注射PDTC 25 mg·kg-1)、PDTC+顺铂组（Lewis细胞+腹腔注射顺铂和PDTC）,每组10只。末次给药后,生化分析仪检测血清肌酐（SCr）、尿素氮（BUN）、胱抑素C（Cys C）、尿液肾损伤分子-1（Kim-1）水平,ELISA法测定肾组织中超氧化物歧化酶（SOD）和丙二醛（MDA）水平,RT-PCR检测肾组织肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-1β、IL-6水平,Western Blotting检测核因子2相关因子2（Nrf2）、血红素氧合酶-1（HO-1）、NF-κB p65、p-IκBα/IκBα的表达,HE染色检测肾组织损伤。结果:与肺癌组相比,顺铂组大鼠血SCr、BUN、Cys C、尿Kim-1水平及肾组织MDA、TNF-α、IL-1β、IL-6 mRNA、Nrf2、HO-1、NF-κB p65和p-IκBα/IκBα的表达明显升高（P<0.05）,肾组织SOD活性和GSH-Px活性明显降低（P<0.05）;与顺铂组相比,PDTC+顺铂组大鼠血SCr、BUN、Cys C、尿Kim-1水平及肾组织MDA、TNF-α、IL-1β、IL-6 mRNA、NF-κB p65和p-IκBα/IκBα的表达明显明显降低（P<0.05）,肾组织SOD、GSH-Px、Nrf2和HO-1的表达明显升高（P<0.05）。结论:抑制NF-κB信号通路可以降低炎症因子水平,还可以激活Nrf2的表达,降低氧化应激水平,保护顺铂致肺癌大鼠的肾组织损伤。     

清开灵注射液对大鼠阳离子化牛血清白蛋白肾炎的作用机制研究

目的探讨清开灵注射液(QKL)对实验性大鼠肾炎的作用机制。方法采用牛血清白蛋白(C-BSA)建立肾炎大鼠模型,QKL腹腔注射治疗,分别取大鼠肾皮质和全血,检测NF-кB含量表达,IL-2、IL-6和TNF-α含量测定。结果与正常对照组相比,模型组的NF-кB表达显著增高(P<0.01),IL-2、IL-6和TNF-α含量显著增高(P<0.05);与模型组相比,清开灵治疗组NF-кB表达,IL-2、IL-6和TNF-α含量则明显降低(P<0.05)。结论清开灵注射液通过抑制NF-кB的活化,降低炎症免疫细胞因子表达,起到治疗肾小球肾炎的作用,因而从分子水平揭示了清热解毒药物治疗肾小球肾炎的部分机制。     

Genipin alleviates LPS-induced acute lung injury by inhibiting NF-κB and NLRP3 signaling pathways

Genipin has been reported to have anti-inflammatory effect. However, its role on lipopolysaccharide (LPS)-induced acute lung injury (ALI) has not been explored. This study aimed to evaluate the effect of genipin on murine model of acute lung injury induced by LPS. The mice were treated with genipin 1 h before LPS administration. 12 h later, the myeloperoxidase (MPO) in lung tissues and lung wet/dry ratio were detected. The levels of TNF-α, IL-1β and IL-6 in bronchoalveolar lavage fluid (BALF) were measured by ELISA. Apart from this, we use western blot to detect the protein expression in the NF-κB and NLRP3 signaling pathways. The results showed that the treatment of genipin markedly attenuated the lung wet/dry ratio and the MPO activity. Moreover, it also inhibited the levels of TNF-α, IL-1β, IL-6 in the BALF. In addition, genipin significantly inhibited LPS-induced NF-κB and NLRP3 activation. In conclusion, these results demonstrate that genipin protected against LPS-induced ALI through inhibiting NF-κB and NLRP3 signaling pathways.     

Fraxin ameliorates lipopolysaccharide-induced acute lung injury in mice by inhibiting the NF-κB and NLRP3 signalling pathways

Fraxin, the effective component of the Chinese traditional medicine Cortex Fraxini, is reported to have anti-inflammatory effects. This study assessed the anti-inflammatory effect of fraxin on the lipopolysaccharide (LPS)-induced inflammatory response in A549 cells and the protective efficacy on LPS-induced acute lung injury (ALI) in mice. Fraxin reduced LPS-induced TNF-α, IL-6 and IL-1β production in A549 cells and alleviated the LPS-induced wet/dry (W/D) weight ratio and the effects observed via histopathological examination of the lung in vivo. Furthermore, fraxin reduced the protein concentrations in the broncho-alveolar lavage (BAL) fluid and cytokine production in the sera. Fraxin also clearly attenuated the oxidation index, including the activity of myeloperoxidase (MPO), malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH). Immunohistochemistry analysis showed that fraxin suppressed LPS-induced inflammatory damage. The expression of proteins involved in the NF-κB and NLRP3 inflammatory corpuscle signalling pathways was consistent between the lung tissues and cell samples. Overall, fraxin played a protective role in LPS-induced lung injury by inhibiting the NF-κB and NLRP3 signalling pathways.     

他克莫司联合雷公藤多苷对肾病综合征患者NF-κB与炎症细胞因子的影响

目的：探讨他克莫司联合雷公藤多苷对肾病综合征患者核转录因子kappa B（NF-κB）与炎症细胞因子（IL-1、TNF-α）的影响。方法：选取我院肾病综合征患者129例,采用随机数字表法将其分为两组,对照组64例,采用他克莫司治疗;研究组65例,采用他克莫司联合雷公藤多苷治疗。采用Elisa方法检测血清NF-κB、IL-1、TNF-α水平,采用酶动力法测定血肌酐（Scr）,采用尿素酶速率法测定血尿素氮（BUN）,比较两组的临床疗效。结果：研究组患者临床缓解率明显高于对照组（P<0.05）。治疗后,全部患者NF-κB、IL-1、TNF-α水平明显低于治疗前,治疗前后比较差异具有统计学意义（P<0.05）,其中研究组NF-κB、IL-1、TNF-α水平明显低于对照组（P<0.05）,治疗后,全部患者Scr、BUN、24 h Upro水平明显低于治疗前（P<0.05）,其中研究组Scr、BUN、24 h Upro水平明显低于对照组（P<0.05）。两组患者药物不良反应率比较差异无统计学意义（P>0.05）。结论：他克莫司联合雷公藤多苷显著降低肾病综合征患者NF-κB与IL-1、TNF-α炎症细胞因子,改善肾功能,且药物安全性较高。     

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.     

Erythropoietin attenuates cardiopulmonary bypass-induced renal inflammatory injury by inhibiting nuclear factor-kappa B P65 expression

Acute renal injury is one of the most frequent complications after cardiopulmonary bypass (CPB). This study was designed to evaluate the potential protective effect of erythropoietin (EPO) on CPB-induced renal injury in a rat model. Male Sprague-Dawley rats were randomly divided into three groups, sham-operated group (sham), control CPB group (control), erythropoietin CPB group (EPO). Blood samples were collected at the beginning, at the end of CPB, and at 0.5, 1, 2 and 24 h post-operation, and the kidneys were harvested 24 h postoperatively and observed by optical microscopy. Levels of serum creatinine (Cr) and blood urea nitrogen (BUN) were assayed. Tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6(IL-6) levels in the renal tissues were evaluated by the method of enzyme linked immunosorbent assay (ELISA). Protein and mRNA levels of nuclear factor kappa B p65 (NF-κB p65), intercellular adhesion molecule-1 (ICAM-1) were also determined using western blot and real-time PCR respectively. Serum Cr and BUN levels as well as TNF-α, IL-1β and IL-6 levels in renal tissues in control group were significantly higher than those in the sham group. However, the levels of above biomarkers were markedly decreased in EPO group when comparing with control group. Furthermore, NF-κB p65, ICAM-1 protein and mRNA expression were significantly down-regulated in EPO group comparing with control group. In addition, microscopic examinations revealed that histological injury was alleviated when treated with EPO. The results indicated that EPO potently protected against CPB-induced acute renal injury and inhibited expression of NF-κB p65 and inflammatory response.     

Therapeutic efficacy of carboxyamidotriazole on 2,4,6-trinitrobenzene sulfonic acid-induced colitis model is associated with the inhibition of NLRP3 inflammasome and NF-κB activation

Excess proinflammatory cytokines owing to the activation of NF-κB and NLRP3 inflammasome play the key role in inflammatory bowel disease (IBD). Previously, we reported the anti-inflammatory activity of carboxyamidotriazole (CAI) resulting from decreasing cytokines. Therefore, we investigated the therapeutic effects of CAI in 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced rat colitis and the involvement of CAI action with NLRP3 inflammasome and NF-κB pathway. CAI was orally administered to TNBS-induced colitis rat. The severity of colitis was assessed, and NLRP3 inflammasome, NF-κB pathway and cytokines were determined. Our results showed that CAI significantly reduced weight loss and disease activity index (DAI) scores in colitis rats and alleviated the colonic macroscopic signs and pathological damage. In addition, the intestinal inflammatory markers and permeability index were markedly ameliorated by CAI treatment. The decreased levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6, IL-18 were also detected in the colon tissues of CAI-treated colitis rats. Moreover, the activation of NLRP3 inflammasome in inflamed colon was significantly suppressed by showing an obvious reduction in the NLRP3 and activated caspase-1 levels. Furthermore, CAI reduced NF-κB p65 expression and IκBα phosphorylation and degradation in colitis rats. Therefore, CAI attenuates TNBS-induced colitis, which may be attributed to its inhibition of NLRP3 inflammasome and NF-κB activation, and down-regulation of proinflammatory cytokines. These results provide further understanding of the intestinal anti-inflammatory effect of CAI and highlight it as a potential drug for the treatment of IBD.     

新型吡唑并[4,3-d]嘧啶衍生物的合成和抗炎活性

目的为了研发新型抗炎药,设计合成了一种新的吡唑并[4,3-d]嘧啶衍生物,并揭示了新化合物可能的抗炎机制。方法在显微镜下观察细胞的形态变化;通过Griess法测定新型化合物对脂多糖刺激的RAW264.7细胞中NO产生的影响;通过qRT-PCR评估TNF-α、IL-6和IL-1β的基因相对转录水平;通过qRT-PCR和Western blot测量环氧化酶-2表达,以及对NF-κB/NLRP3炎症小体信号通路进行检测。小鼠体内实验通过HE染色观察肺组织变化。结果该新型化合物借助NF-κB/NLRP3炎症小体信号通路可有效抑制RAW264.7细胞中NO的产生以及COX-2、TNF-α、IL-6和IL-1β的表达。同时,它还可以改善细胞的形态和缓解小鼠急性肺损伤。结论新型吡唑并[4,3-d]嘧啶衍生物通过NF-κB/NLRP3炎症小体信号通路来产生抗炎活性。     

激动黑皮质素4型受体对抗脓毒症诱导的肾脏损伤

目的:观察激动黑皮质素4型受体(MC4R)对抗脓毒症诱导的肾脏损伤作用,并探讨其机制.方法:雄性SD大鼠24只,随机分为:假手术组(Sham)、Sham+MC4R激动剂R027-3225组、盲肠结扎穿孔组(CLP)和CLP+R027-3225组,造模24 h后测定血清尿素氮(BUN)和肌酐(CRE)水平,HE染色观察肾脏形态学变化,免疫组化观察肾小管细胞NF-κB P65的表达,RT-PCR测定肾脏组织NF-κB mRNA表达情况.结果:与Sham组相比,CLP组血清BUN和CRE水平明显升高(P＜0.01),组织切片显示CLP组大鼠肾小体中血管球淤血、皱缩、肾小囊腔扩大,伴肾小管上皮细胞肿胀和坏死.肾脏组织NF-κB P65表达呈强阳性,NF-κB mRNA表达增高(P＜0.01).与CLP组相比,CLP+R027-3225组BUN和CRE水平下降(P＜0.01),肾小体、肾小管损伤减轻,肾脏组织NF-κB P65表达和NF-κB mRNA表达下降(P＜0.01).结论:脓毒症大鼠肾脏功能受损,激动MC4R可对抗脓毒症诱导的肾脏损伤,这可能与减少NF-κB的表达,降低炎症反应有关.