盐酸戊乙奎醚预先给药对脓毒症大鼠肺组织高迁移率族蛋白B1表达的影响

Effects of Penehyclidine Hydrochloride Pretreatment on Expression of High Mobility Group Box 1 in Lung Tissue in Septic Rats

目的评价盐酸戊乙奎醚预先给药对脓毒症大鼠肺组织高迁移率族蛋白B1(HMGB1)表达的影响。方法健康雄性SD大鼠108只,4~5月龄,体质量300~350 g,采用随机数字表法分为:假手术组(S组)、脓毒症模型组(CLP组)和盐酸戊乙奎醚+脓毒症模型组(PH组),每组36只。CLP组和PH组采用盲肠结扎穿孔(CLP)制备脓毒症大鼠模型。PH组于模型制备前1 h时静脉注射盐酸戊乙奎醚0.1 mg·kg^(-1),S组和CLP组给予等容量生理盐水。分别于术前2 h(TO)、术后2、12、24、48和72 h(T1~T5)时随机取6只大鼠,收集支气管肺泡灌洗液(BALF),采用考马斯亮兰法检测BALF总蛋白;测定肺组织湿/干质量比;采用分光光度法检测肺组织髓过氧化物酶(MPO)活性;采用Elisa和RT-PCR检测肺组织HMGB1含量和mRNA表达水平。结果与S组比较,CLP组肺组织湿/干质量比、BALF总蛋白含量、MPO活性、HMGB1含量和HMGB1 mRNA表达升高(P<0.05);与CLP组比较,PH组肺组织湿/干质量比、BALF总蛋白含量、MPO活性、HMGB1含量和HMGB1 mRNA表达降低(P<0.05)。结论预先给药盐酸戊乙奎醚可下调脓毒症大鼠肺组织中HMGB1表达,这可能是其减轻肺损伤的一个重要机制。

Objective To investigate the effects of penehyclidine hydrochloride pretreatment on the expression of high mobility group box 1（HMGB1）in the lung tissue in septic rats.Methods One hundred and eight male SD rats weighting 300-350 g（4-5 months old）were randomly divided into three groups：sham operation group（group S）,sepsis group（group CLP）and penehyclidine hydrochloride treatment group（group PH）,with 36 rats in each group.Sepsis was induced by ce-cal ligation and puncture（CLP）in groups CLP and PH.Rats in group PH were intravenously in-jected with 0.1 mg/kg penehyclidine hydrochloride 1 hour before CLP.Rats in groups S and CLP were given the same volume of normal saline.Six animals in each group were killed 2 hours before CLP and 2,12,24,48 and 72 hours after CIP,respectively.Bronchoalveolar lavage fluid（BALF） was collected to examine the protein content.The lung tissue samples were collected to examine the lung wet/dry weight ratio.Myeloperoxidase（MPO）activity in the lung tissue was determined by spectrophptometry.Protein and mRNA expression of HMGB1 in the lung tissue was detected＆nbsp;by ELISA and real-time PCR,respectively.Results Compared with group S,the lung wet/dry weight ratio,BALF protein content,MPO activity,and HMGB1 protein and mRNA expression significantly increased in group CLP（P 〈 0.05）.Compared with group CLP,the lung wet/dry weight ratio,BALF protein content,MPO activity,and HMGB1 protein and mRNA expression significantly decreased in group PH（P 〈0.05）.Conclusion PHC pretreatment can attenuate the lung injury induced by sepsis in rats through down-regulating the expression of HMGB1.