摘要
目的 探讨自噬在脓毒症小鼠急性肺损伤中的作用.方法 雄性C57BL/6小鼠36只,6周龄,体重20~ 25 g,采用随机数字表法,将其分为3组(n=12):假手术组(S组)、脓毒症组(CLP组)和脓毒症+自噬抑制剂3-甲基腺嘌呤组(CLP+3-MA组).采用盲肠结扎穿孔法制备脓毒症模型.CLP+3-MA组术后1h时腹腔注射3-甲基腺嘌呤10 mg/kg.术后24 h时取颈动脉血样,行血气分析,计算氧合指数;取肺组织,行肺组织病理学评分,测定肺组织湿重/干重比值(W/D比值),采用比色法测髓过氧化酶(MPO)活性,采用Western blot法测定自噬蛋白微管相关蛋白1轻链3Ⅱ型(LC3Ⅱ)、Beclin-1和溶酶体相关蛋白Rab7和LAMP2的表达,进行支气管肺泡灌洗,计数支气管肺泡灌洗液(BALF)总细胞和多形核中性粒细胞(PMN).结果 与S组比较,CLP组和CLP +3-MA组肺组织病理学评分、W/D比值、MPO活性、BALF总细胞计数和PMN计数升高,氧合指数降低,CLP组肺组织LC3Ⅱ、Beclin-1、LAMP2和Rab7的表达上调(P<0.05).与CLP组比较,CLP+3-MA组肺组织病理学评分、W/D比值、MPO活性、BALF总细胞计数和PMN计数升高,氧合指数降低,肺组织LC3Ⅱ、Beclin-1、LAMP2和Rab7的表达下调(P<0.05).结论 自噬参与了脓毒症小鼠急性肺损伤时的内源性保护机制.
Objective To investigate the role of autophagy in the lung injury in the septic mice.Methods Thirty-six male C57BL/6 mice, aged 6 weeks, weighing 20-25 g, were randomly divided into 3 groups (n=12 each) using a random number table: sham operation group (group S);cecal ligation and puncture (CLP) group;CLP + autophagy inhibitor 3-methyladenine (3-MA) group (group CLP+3-MA).Sepsis was produced by CLP.In group CLP+3-MA, 3-MA 10 mg/kg was injected intraperitoneal at 1 h after operation.Arterial blood samples were taken at 24 h after operation for blood gas analysis, and the oxygenation index was calculated.The lungs were removed for microscopic examination of pathologic changes which were scored, and for determination of wet/dry lung weight ratio (W/D ratio) , myeloperoxidase (MPO) activity (using colorimetric method) and the expression of autophagy protein microtubule-associated protein 1 light chain 3 Ⅱ] (LC3 Ⅱ), Beclin-1 and lysosomes-associated protein Rab7 and lysosome-associated membrane protein-2 (LAMP2) (by Western blot).The lung was lavaged, and broncho-alveolar lavage fluid (BALF) was collected for determination of the total cell count and polymorphonuclear leukocyte (PMN) count.Results Compared with group S, the pathological score, W/D ratio, MPO activity, and the total cell and PMN counts in BALF were significantly increased, and oxygenation index was decreased in CLP and CLP +3-MA groups, and the expression of LC3 Ⅱ , Beclin-1, LAMP2 and Rab7 was up-regulated in group CLP (P〈0.05).Compared with group CLP, the pathological score, W/D ratio, MPO activity, and the total cell and PMN counts in BALF were significantly increased, the oxygenation index was decreased, and the expression of LC3 Ⅱ , Beclin-1, LAMP2 and Rab7 was down-regulated in group CLP+ 3-MA (P〈0.05).Conclusion Autophagy is involved in the endogenous protective mechanism of acute lung injury in the septic mice.
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2015年第9期1124-1127,共4页
Chinese Journal of Anesthesiology
基金
国家自然科学基金(81372033,81471842)
天津市自然科学基金(13JCQNJC11400)
关键词
自噬
脓毒症
呼吸窘迫综合征
成人
Autophagy
Sepsis
Respiratory distress syndrome,adult