中文摘要：

【目的】探讨注射钴原卟啉（cobalt protoporphyrin,Co PP）在内毒素诱导的急性肺损伤（acute lung injury,ALI）小鼠模型中上调血红素加氧酶-1（heme oxygenase-1,HO-1）对小鼠肺组织炎症损伤的影响。【方法】120只16~18 g雄性C57BL/6小鼠随机分为生理盐水（normal saline,NS）组、脂多糖（lipopolysaccharide,LPS）组、LPS＋NS组以及LPS＋钴原卟啉（Co PP）组。采取经腹腔注射LPS（5 mg/kg）法建立实验性ALI模型。LPS＋Co PP组及LPS＋NS组均在注射LPS造模之前4 h,分别预先经腹腔注射给予Co PP（5 mg/kg）或等体积的无菌NS。在1、3、7天后分别取材,采用HE染色观察肺泡、肺间质炎症浸润情况并进行炎症评分（inflammatory score,IS）;采用BCA蛋白定量法检测支气管肺泡灌洗液（bronchoalveolar lavage fluid,BALF）中总蛋白含量,ELISA检测BALF及血清中白介素-8（interleukin 8,IL-8）、单核细胞趋化蛋白-1（monocyte chemoattractant protein,MCP-1）含量;使用乳酸盐脱氢酶（lactate dehydrogenase,LDH）试剂盒用微板法检测BALF中LDH活性;右肺分别提取总RNA或总蛋白,采用RTPCR联合2-△△Ct计算方法检测评估HO-1 m RNA相对表达量,Western blotting检测髓过氧化物酶（myeloperoxidase,MPO）蛋白表达水平。【结果】在第1天,LPS＋Co PP组HO-1 m RNA水平均明显高于LPS＋NS组（P〈0.01）,在第3天持续升高（P〈0.01）,第7天各组无差异。与NS组相比,LPS组及LPS＋NS组小鼠第1天的IS、BALF总蛋白含量和LDH活性及BALF和血清中IL-8、MCP-1水平均明显增高;在第3天持续升高,且肺组织MPO蛋白表达量亦增高（P〈0.01）。与LPS＋NS组相比,LPS＋Co PP组小鼠第1天的IS、BALF总蛋白含量、LDH活性及BALF和血清中IL-8水平均降低,在第3天除以上4个指标持续降低外,BALF及血清中MCP-1、肺组织MPO蛋白表达水平也均降低（P〈0.01）。【结论】Co PP上调HO-1 m RNA的表达,可能是通过下调炎性趋化因子IL-8和MCP-1

英文摘要：

【Objective】To explore the effect of heme oxygenase-1（HO-1） up-regulated by cobalt protoporphyrin（Co PP） on inflammatory injury of mouse lung tissues in models of lipopolysaccharide（LPS）-induced acute lung injury（ALI）.【Methods】A total of 120 C57BL/6 mice（body weight 16~18 g） were randomly divided into NS group, LPS group, LPS＋NS group and LPS＋Co PP group.Experimental ALI models were made by intraperitoneal injection of LPS（5mg/kg）. Mice in LPS＋Co PP group and LPS＋NS group were injected with Co PP（5 mg/kg） or the same volume of NS at the 4 h before LPS injection. After 1, 3 or 7 days, HE staining was used to observe alveolar and interstitial inflammation, and inflammatory score（IS） was calculated. Total protein quantification of in BALF supernatant was detected with BCA protein assay kits. The levels of IL-8 and MCP-1 in the BALF and serum were measured by ELISA kits. LDH activity was determined by LDH activity kit. Total RNA and protein were respectively isolated from non-lavaged right lung tissues. The relative HO-1 m RNA expression level was normalized with RT-PCR and 2-△△Ctmethod. The protein expression level of MPO was determined by Western blotting.【Results】The expression level of HO-1 m RNA in LPS ＋ Co PP group was significantly higher than that in LPS ＋ NS group on the 1st day（P〈0.01） and rose continuously on the 3rd day（P〈0.01）. There was no significant difference in m RNA expression level of HO-1 on the 7th day. Compared to NS group, the IS, total protein content in BALF, LDH activity, levels of IL-8 and MCP-1 in BALF and serum all increased significantly in LPS group and LPS＋NS group on the 1st day, and rose continuously on the 3rd day. Moreover, the protein expression of MPO in lung tissues was significantly up-regulated on the 3rd day（P〈0.01）. Compared with LPS＋NS group, the IS, total protein content in BALF, LDH activity, IL-8 level in BALF and serum in LPS＋Co PP group all decreased on the 1st day, and MCP-1 levels in