中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
Chinese Journal of Anesthesiology
2015年
9期
1124-1127
,共4页
董艾莉%陈红光%谢克亮%于泳浩
董艾莉%陳紅光%謝剋亮%于泳浩
동애리%진홍광%사극량%우영호
自噬%脓毒症%呼吸窘迫综合征,成人
自噬%膿毒癥%呼吸窘迫綜閤徵,成人
자서%농독증%호흡군박종합정,성인
Autophagy%Sepsis%Respiratory distress syndrome,adult
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.
目的 探討自噬在膿毒癥小鼠急性肺損傷中的作用.方法 雄性C57BL/6小鼠36隻,6週齡,體重20～ 25 g,採用隨機數字錶法,將其分為3組(n=12):假手術組(S組)、膿毒癥組(CLP組)和膿毒癥+自噬抑製劑3-甲基腺嘌呤組(CLP+3-MA組).採用盲腸結扎穿孔法製備膿毒癥模型.CLP+3-MA組術後1h時腹腔註射3-甲基腺嘌呤10 mg/kg.術後24 h時取頸動脈血樣,行血氣分析,計算氧閤指數;取肺組織,行肺組織病理學評分,測定肺組織濕重/榦重比值(W/D比值),採用比色法測髓過氧化酶(MPO)活性,採用Western blot法測定自噬蛋白微管相關蛋白1輕鏈3Ⅱ型(LC3Ⅱ)、Beclin-1和溶酶體相關蛋白Rab7和LAMP2的錶達,進行支氣管肺泡灌洗,計數支氣管肺泡灌洗液(BALF)總細胞和多形覈中性粒細胞(PMN).結果 與S組比較,CLP組和CLP +3-MA組肺組織病理學評分、W/D比值、MPO活性、BALF總細胞計數和PMN計數升高,氧閤指數降低,CLP組肺組織LC3Ⅱ、Beclin-1、LAMP2和Rab7的錶達上調(P＜0.05).與CLP組比較,CLP+3-MA組肺組織病理學評分、W/D比值、MPO活性、BALF總細胞計數和PMN計數升高,氧閤指數降低,肺組織LC3Ⅱ、Beclin-1、LAMP2和Rab7的錶達下調(P＜0.05).結論 自噬參與瞭膿毒癥小鼠急性肺損傷時的內源性保護機製.
목적 탐토자서재농독증소서급성폐손상중적작용.방법 웅성C57BL/6소서36지,6주령,체중20～ 25 g,채용수궤수자표법,장기분위3조(n=12):가수술조(S조)、농독증조(CLP조)화농독증+자서억제제3-갑기선표령조(CLP+3-MA조).채용맹장결찰천공법제비농독증모형.CLP+3-MA조술후1h시복강주사3-갑기선표령10 mg/kg.술후24 h시취경동맥혈양,행혈기분석,계산양합지수;취폐조직,행폐조직병이학평분,측정폐조직습중/간중비치(W/D비치),채용비색법측수과양화매(MPO)활성,채용Western blot법측정자서단백미관상관단백1경련3Ⅱ형(LC3Ⅱ)、Beclin-1화용매체상관단백Rab7화LAMP2적표체,진행지기관폐포관세,계수지기관폐포관세액(BALF)총세포화다형핵중성립세포(PMN).결과 여S조비교,CLP조화CLP +3-MA조폐조직병이학평분、W/D비치、MPO활성、BALF총세포계수화PMN계수승고,양합지수강저,CLP조폐조직LC3Ⅱ、Beclin-1、LAMP2화Rab7적표체상조(P＜0.05).여CLP조비교,CLP+3-MA조폐조직병이학평분、W/D비치、MPO활성、BALF총세포계수화PMN계수승고,양합지수강저,폐조직LC3Ⅱ、Beclin-1、LAMP2화Rab7적표체하조(P＜0.05).결론 자서삼여료농독증소서급성폐손상시적내원성보호궤제.
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.
