白藜芦醇对脓毒症大鼠NF-κB/IκB调节机制的研究

Study on the regulation mechanism of resveratrol on NF-κB/IκB in sepsis

目的探讨白藜芦醇对脓毒症核转录因子κB(NF-κB)/IκB调节炎症的机制。方法以盲肠结扎穿刺法(Clp)建立大鼠脓毒症模型,30只SD大鼠按随机数字表法分成3组,每组10只。假手术组(Sham组):麻醉后作腹部正中2 cm切口,不做其他操作,随即关腹。脓毒症组(Clp组):麻醉后作腹部正中2 cm切口,从腹腔找出盲肠后结扎其1/2长度,用20 G针贯穿两次,挤出粪便然后将盲肠放回腹腔,随即关腹。白藜芦醇组(Res+Clp组):术前5 d腹腔注射白藜芦醇,每天一次,用量15 mg/kg,其余操作均与Clp组相同。所有大鼠术毕皮下注射生理盐水30 ml/kg复苏。观察18 h肾脏病理、肾功能改变;使用Western Blot技术检测肾组织NF-κB Pp65、IKK-β和IκBα的表达。结果 Sham组肾脏结构正常;Clp组可见部分肾小管上皮细胞变性、坏死、脱落和管型形成;Clp+Res组可见肾小管扩张和管型减少。Clp组尿素氮(BUN)和肌酐(Cr)值明显高于Sham组(P<0.05);Clp+Res组与Clp组相比,Clp+Res组BUN和Cr值明显降低(P<0.05)。Clp组与Sham组相比Pp65蛋白表达量明显增加(P<0.05);Clp+Res组Pp65的表达量相对于Clp组明显减少(P<0.05)。Clp组的IKK-β蛋白表达量较Sham组显著增加(P<0.05);Clp+Res组的IKK-β表达量比Clp组明显减少(P<0.05)。Clp组的IκB蛋白表达量较Sham组显著减少(P<0.05);Clp+Res组的IκB表达量比Clp组明显增加(P<0.05)。结论白藜芦醇能通过抑制IKK-IκB-Pp65炎症通路减轻脓毒症大鼠肾损伤。

Objective To study the regulation mechanism of resveratrol on NF-κB/IκB in sepsis. Methods The septic model were set up by cecal ligation and puncture（ Clp）,thirty rats were randomly divided into three groups in ten of each group including Sham group,Clp group,and resveratrol group. Sham operation group（ Sham group） ： after anesthesia,the 2 cm incision in the middle of the abdomen was done without any other operation,then the abdomen was closed. Sepsis group（ Clp group） ： after anesthesia,abdominal median 2 cm incision was performed. After the cecum was found,the 1/2 length was ligated,and the 20 G needle was passed through the cecum two times. The stool was then squeezed out,and the cecum was put back to the abdominal cavity,then the abdomen was closed. Resveratrol group（ Res ＋ Clp group） ： 5 days before the operation,resveratrol was injected intraperitoneally,once a day,the amount of 15 mg/kg was used,and the rest of the process was the same as that of the Clp group. All rats were injected with saline Bipi under 30 ml/kg recovery. The kidney pathological changes,the level of BUN,Cr,NF-κB Pp65,IKK-βand IκBα. Results In Sham group,the renal structure was normal. In group Clp,partial renal tubular epithelial cells were denatured,necrotic,shedding and tubular formation. Clp ＋ Res group showed renal tubular dilatation and tubular formation reduction. The value of BUN and Cr in group Clp was significantly higher than that in group Sham（ P〈0. 05）,and in Clp ＋ Res group,the BUN and Cr values of Clp ＋ Res group were significantly lower than those in Clp group（ P〈0. 05）. The expression of the Pp65 protein in group Clp was significantly increased compared with that in group Sham（ P〈0. 05）,and the expression of Pp65 in group Clp ＋ Res was markedly lower than that in group Clp（ P〈0. 05）. The expression of IKK-beta protein in group Clp was significantly higher than that in group Sham（ P〈0. 05）,and the expression of IKK-beta in group Clp ＋ Res was considerably lower than that in group Clp（ P〈0. 05）. The expression of I kappa B protein in group Clp was significantly lower than that in group Sham（ P〈0. 05）,and the expression of I kappa B in group Clp ＋ Res was markedly higher than that in group Clp（ P〈0. 05）. Conclusion Resveratrol could alleviate renal impairment through inhibiting NF-κB/IκB pathway in sepsis.