Shp-2/NF-κB途径介导脂氧素A4对IL-1β促肾小球系膜细胞IL-6合成作用的抑制

目的 :研究脂氧素A4 (LXA4 )是否拮抗IL 1β促进肾小球系膜细胞IL 6合成的作用 ,并探讨LXA4 作用的信号转导机制。方法 :用不同浓度的LXA4 预处理培养的大鼠肾小球系膜细胞 ,再加入IL 1β共同孵育一定时间 ;或单用IL 1β刺激肾小球系膜细胞。用ELISA法检测培养上清中的IL 6蛋白表达量 ;用RT PCR法检测IL 6的mRNA表达量 ;用免疫沉淀与印迹法检测含Src同源区 2 (SH2 )蛋白酪氨酸磷酸酶 - 2 (Shp 2 )的表达 ;用凝胶电泳迁移率试验 (EMSA)检测NF κB的DNA结合活性。结果 :LXA4 呈剂量依赖性地抑制IL 1β诱导的系膜细胞分泌的IL 6蛋白及mRNA表达 ,拮抗IL 1β诱导的Shp 2磷酸化与NF κB的活化。 结论 :LXA4 能够拮抗IL 1β对肾小球系膜细胞IL 6合成的促进作用 ,其机制依赖于Shp 2 NF κB信号转导途径     

LXA4对ANP诱发SIRS过程中NF-κB、ICAM-1表达的影响

目的探讨脂氧素A4抑制急性坏死性胰腺炎诱发SIRS的机制。方法健康雄性SD大鼠72只随机分为假手术组、ANP组和脂氧素A4(LXA4)干预组,分别采用等量生理盐水、LXA4(0.1mg/kg)间隔8h经阴茎背静脉注射,并分别于4、12、24h每组活杀8只大鼠,测定胰腺组织Schmidt病理评分、NF-ΚB、ICAM-1的表达水平。结果 (1)与假手术组比较,胰腺炎组、LXA4干预组的Schmidt胰腺病理评分有显著升高(P<0.01);LXA4干预组Schmidt胰腺病理评分在12、24h较胰腺炎组低(P<0.05)。(2)假手术组ICAM-1表达阴性,胰腺炎组、LXA4干预组随着病变进展,其反映ICAM-1表达相对强度的累积光密度值(IOD)逐渐上升(P<0.01);在4、12、24h等3个时点,LXA4干预组的IOD值较比SAP组均有显著下降(P<0.01)。(3)假手术组NF-κB P65蛋白阴性;ANP组NF-κB胞核阳性表达率为59.1%(13/22),显著高于LXA4干预组的4.3%(1/23),差异具有统计学意义(P<0.01)。结论 (1)LXA4在ANP诱发SIRS过程中确能抑制ICAM-1的表达。(2)LXA4抑制ANP诱发SIRS可能是通过抑制NF-κB的细胞信号传导途径来实现的。     

祛风通络方抑制NF-κB活化对脂多糖诱导的体外培养大鼠肾小球系膜细胞TGF-β1mRNA和IL-6mRNA表达的影响

目的 研究祛风通络方对脂多糖刺激大鼠肾小球系膜细胞表达炎性细胞因子TGF-β1、IL-6的影响及其作用机制.方法 采用体外培养大鼠系膜细胞和血清药理学方法,分为空白组,病理组,治疗组和阳性对照组.采用生物素核酸一蛋白结合凝胶迁移电泳分析法检测NF-KB的表达;RT-PCR检测TGF-β1mRNA和IL-6mRNA表达.结果 祛风通络方能够抑制脂多糖刺激肾小球系膜细胞中NF-KB活化,且在祛风通络方的作用下,TGF-β1mRNA和IL-6mRNA表达均呈不同程度减弱.结论 提示祛风通络方可能通过抑制NF-B活化,下调TGF-131mRNA和IL-6mRNA的表达达到减轻系膜细胞增殖,防止肾小球硬化的作用.这可能是祛风通络方治疗系膜增生性肾炎的机制之一.     

Expression and Significance of NF-κB, IL-1β and COX-2 in the Murine Model of Estrogen-dependent Experimental Vulvovaginal Candidiasis

Objective To investigate the possible role of estrogen in the pathogenesis of vulvovaginal candidiasis(VVC). Methods Estrogen-dependent experimental murine model of C. albicans vaginal infection was established by injecting subcutaneously with estradiol benzoate and then 5×106 stationary-phase C. albicans blastoconidia was inoculated intravaginally to mice (group EI), and other 3 groups were set up: estrogen-treated but not infected (group E); estrogen-untreated but infected (group I); normal control (group C). The dynamic change of colony-forming unit (CFU) of cervivovaginal lavage fluid was observed. Vaginal tissues at different time points (d 2, d 4, d 7 and d 14) after inoculation of C. albicans were obtained. In situ hybridization staining was used to detect expression of cyclooxygenase-2 (COX-2) mRNA and expression of nuclear factor-κB (NF-κB) was examined by immuohistochemistry. ELISA was applied to determine the interlenkin-1β (IL-1β) level. Results The constitutional high level expression of COX-2 mRNA in the vaginal tissue of group E was significantly higher than that in group C on d 4 and d 7 (P<0.01), and the optical density (OD) in group EI was higher than that in the other 3 groups (P<0.05). There were higher IL-1a levels in vaginal tissues from d 4 to d 7 postin- oculation in group EI and group I than group C (P<0.01). Furthermore, IL-1β in group EI was markedly elevated on d 4 and d 7 compared with group I (P<0.01). Compared with group C, the expression of NF-κB in group E was increased obviously on d 4 (P<0.01), and there was significant difference between group EI and group Ion d 4 and d 7 (P<0.01). Conclusions In the murine model of estrogen-dependent experimental VVC, estrogen promotes the infection establishment by up-regulating expression of COX-2 via activating NF-κB signal pathway, and the high expression of COX-2 promoted by the interaction of IL-1β and NF-êB after infection formation was involved in persistence of infection.     

雌激素依赖的外阴阴道假丝酵母菌病小鼠模型中NF-κB、IL-1β和COX-2 mRNA的表达及意义

目的观察NF-κB、IL-1β和COX-2 mRNA在雌激素依赖的外阴阴道假丝酵母菌病(VVC)小鼠模型中的动态变化,探讨雌激素诱发VVC的可能机制。方法采用雌性昆明种小鼠,预先皮下注射雌激素使其处于假发情状态,然后接种5×106假丝酵母菌孢子构建VVC模型(EI组),并设置雌激素处理未感染组(E组)、未用雌激素处理感染组(I组)和空白对照组(C组)。动态观察小鼠阴道灌洗液真菌载量计数(CFU),于接种后的不同时间点摘取小鼠阴道组织。分别采用原位杂交、免疫组化和ELISA检测其COX-2 mRNA、NF-κB的表达及IL-1β水平。结果COX-2 mRNA在E组阴道组织的表达水平明显高于C组(P<0.01),EI组表达水平高于其它3组;接种后第4、7、14天EI组及I组IL-1β水平均高于C组(P<0.01),EI组在第4、7天高于I组(P<0.01);E组NF-κB的阳性表达率从第4天开始就明显高于C组(P<0.01),而EI组显著高于I组(P<0.01)。结论在雌激素依赖的VVC小鼠模型中,雌激素激活阴道组织中NF-κB信号通路,上调COX-2的表达,与感染形成有关;而形成感染后,IL-1β和NF-κB相互作用使COX-2强表达则与感染持续有关。