【作者】 李锐； 张彧； 邢静；

【Author】 Zhang yu,Xing jing,Li rui ED,the first affiliated hospital of Dalianmedical university,dalian,china

【机构】 大连医科大学附属第一医院急诊科；

【摘要】 目的:动态观察脓毒症大鼠外周血单核细胞TLR4表达变化及TGF-β₁调节脓毒症的作用及机制。方法:清洁级雄性SD大鼠120只,随机分为对照组（随机分5组,每组8只）、脓毒症模型组及TGF-β₁干预组,（各组再随机分5组,分别为脓毒症2h组、6h组、12h组、24h组、48h组）。脓毒症模型组及TGF-β₁干预组以盲肠结扎穿孔法（cecal ligation puncture,CLP）建立脓毒症模型,脓毒症模型组于造模后0.5小时尾静脉注射生理盐水1ml,TGF-β₁干预组于造模后0.5小时尾静脉注射TGF-β₁（20ng/ml·250g）。利用流式细胞术检测外周血单核细胞表面TLR4表达变化,酶联免疫吸附分析法（ELISA）检测外周血TNF-α浓度变化并于相应时间点活杀大鼠取组织器官观察病理改变。结果:CLP术后2-12h TLR4在外周血单核细胞表面表达逐渐下降,12 h降至最低,此后表达升高,CLP术后6-12h与对照组相比表达明显降低(P<0.05=;TGF-β₁干预组TLR4在单核细胞表达较脓毒症模型组表达降低,CLP造模后2h、24h、48h差异均有统计学意义(P<0.05=,外周血TNF-α浓度较脓毒症模型组明显升高,差异在各时间点均有统计学意义(P<0.05=。TGF-β₁干预组血清INF-α浓度水平与单核细胞表达TLR4阳性率程度不相关。与脓毒症模型组相比,TGF-β₁干预组肾脏、肺脏炎症损伤、组织细胞坏死加剧,在CLP术后24h最为明显。结论:外周血单核细胞TLR4表达下降主要发生在脓毒症后2-12小时内;TGF-β₁能调节TLR4在外周血单核细胞上的表达变化,能促进脓毒症前炎症因子TNF-α生成,加剧肾脏、肺脏炎症损伤,但TGF-β₁调节TNF-α的产生增加不是直接通过调节外周血单核细胞上TLR4表达变化实现的。更多还原

【Abstract】 To investigate the changes of Toll-like receptor 4（TLR4） expression on peripheral blood monocytes and the effect of TGF-βregulating mechanism in rats with sepsis induced by cecal ligation and puncture（CLP）. Methods:A total of 120 adult male SD rats were randomized into 3 groups as follows:normal control group（n=40）,CLP group（n=40） and TGF-β₁ intervention group（n=40）.Rats with sepsis induced by cecal ligation and puncture（CLP）.The percentage of animal’s peripheral blood monocytes TLR4 expression and plasma TNF-αlevels were measured at various intervals（2h、12h、24h、48h after CLP）.Meanwhile,pathological changes in tissue samples from thelungs and kidneys were observed at the same intervals. Results:The expression of TLR4 on peripheral blood monocytes were down-regulated and the significant difference at 6-12h after CLP.Compared with the CLP group,the expression of TLR4 on peripheral blood monocytes was significantly decreased in the TGF-β1 intervention group,there are significant difference at 2h、24h、48h after CLP（P<0.05） and the level of TNF-αsignificantly increased at all intervals in TGF-β₁ intervention group（P<0.05）.Furthermore,no significant correlation was found between plasma TNF-αlcvcls and pcriphcral blood monocytcs TLR4 exprcssion（r= 0.127,P>0.05）.The histomorphology results show that Inflammatory reaction apparently aggravate of lung and renal in CLP group;Compared with CLP group, TGF-β1 intervention group display more severious.Conclusions:The expression of TLR4 on peripheral blood monocytes were down-regulated and mainly at 6-12h after CLP.TGF-β₁ have been show to contribute to the regulation of TLR4 expression on monocytes and the plasma TNF-αlevels,and intensify inflammatory reaction both at lung and renal.But our study evaluate that changes is not directly induced by TLR4 expression.更多还原