小窝蛋白1脚手架区多肽减轻LPS诱导的小鼠急性肺损伤

发布时间：2018-10-20 15:46

【摘要】：目的:探究人工合成的小窝蛋白1(caveolin-1,Cav-1)脚手架区多肽cavtratin对血红素加氧酶1(heme oxygenase-1,HO-1)活性及脂多糖(lipopolysaccharide,LPS)诱导的小鼠急性肺损伤的作用。方法:成年雄性BALB/c小鼠随机分为6组,每组8~10只,实验分为对照(control)组、触足肽内化序列(Antennapedia internalization sequence,AP)组、LPS组、LPS+血晶素(hemin)组、LPS+hemin+cavtratin组和LPS+hemin+cavtratin+锌原卟啉(zinc protoporphyrin IX,Zn PP)组。小鼠气管滴注LPS 24 h后,苏木素-伊红染色观察肺组织病理形态变化;检测肺组织湿/干重比、肺泡灌洗液中细胞数和血清中乳酸脱氢酶活性。免疫荧光观察HO-1和Cav-1的结合情况并检测HO-1活性;实时荧光定量PCR检测炎症因子(IL-1β、IL-6、TNF-α、MCP-1和i NOS)的mRNA水平。结果:组织免疫荧光以及HO-1活性检测发现,与LPS组比较,LPS+hemin+cavtratin组HO-1与细胞膜Cav-1的结合减少,HO-1的活性增高(P0.05);与LPS组比较,LPS+hemin+cavtratin组肺组织受损程度明显减轻,肺湿/干重比、肺泡灌洗液细胞数和血清中乳酸脱氢酶活性显著降低(P0.05);与LPS组比较,LPS+hemin+cavtratin组炎症因子的mRNA表达降低(P0.05);HO-1活性抑制剂Zn PP可以消除cavtratin的保护作用。结论:Cavtratin可以通过减少HO-1与细胞膜Cav-1的结合使得HO-1活性增加,进而减轻LPS诱导的小鼠急性肺损伤。
[Abstract]:Aim: to investigate the effects of synthetic scaffold polypeptide cavtratin of caveolin-1,Cav-1 on heme oxygenase-1 (heme oxygenase-1,HO-1) activity and lipopolysaccharide (lipopolysaccharide,LPS) -induced acute lung injury in mice. Methods: adult male BALB/c mice were randomly divided into 6 groups (n = 10): control (control) group, (Antennapedia internalization sequence,AP group, LPS hemin cavtratin group and LPS hemin cavtratin zinc-protoporphyrin (zinc protoporphyrin IX,Zn PP group. 24 h after LPS was infused into mouse trachea, histopathological changes of lung tissue were observed by hematoxylin eosin staining, lung wet / dry weight ratio, cell number in alveolar lavage fluid and lactate dehydrogenase activity in serum were measured. The binding of HO-1 and Cav-1 and the activity of HO-1 were observed by immunofluorescence, and the levels of IL-1 尾, IL-6,TNF- 伪, MCP-1 and i NOS) mRNA were detected by real-time fluorescence quantitative PCR. Results: compared with LPS group, the binding of HO-1 to cell membrane Cav-1 decreased and the activity of HO-1 increased in, LPS hemin cavtratin group (P0.05), compared with LPS group, the damage degree of lung tissue in, LPS hemin cavtratin group was significantly reduced and lung wet / dry weight ratio was significantly decreased (P < 0.05). The number of alveolar lavage fluid cells and the activity of lactate dehydrogenase in serum decreased significantly (P0.05); compared with LPS group, the mRNA expression of inflammatory factors in, LPS hemin cavtratin group was decreased (P0.05); Zn PP, the inhibitor of HO-1 activity, could eliminate the protective effect of cavtratin. Conclusion: Cavtratin can increase the activity of HO-1 by reducing the binding of HO-1 to cell membrane Cav-1 and then attenuate the acute lung injury induced by LPS in mice.
【作者单位】：江南大学无锡医学院;无锡市第三人民医院急救中心;
【基金】：国家自然科学基金资助项目(No.81270126) 中央高校基本科研业务费专项资金资助(No.JUSRP51412B) 江苏省研究生培养创新工程项目(No.KYZZ16_0312;No.KYLX15_1196) 国家级大学生创新创业训练计划项目(No.201610295074) 江苏省医学会/康缘药业临床医学科研专项基金(No.2015JZKY07)
【分类号】：R563

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