NAD^(+)介导的SIRT1去乙酰化活性在小鼠内毒素性急性肺损伤中的作用  

Role of NAD^(+)-mediated deacetylation activity of SIRT1 in endotoxin-induced acute lung injury in mice

作　　者：何思梦 史佳[2] 李香云 董树安[2] 张圆[2] 余剑波 He Simeng;Shi Jia;Li Xiangyun;Dong Shu′an;Zhang Yuan;Yu Jianbo(School of Medicine,Nankai University,Tianjin 300071,China;Department of Anesthesiology and Critical Care Medicine,Tianjin Nankai Hospital,Tianjin Medical University,Tianjin 300100,China)

机构地区：[1]南开大学医学院,天津300071 [2]天津医科大学南开医院(天津市南开医院)麻醉与重症医学科,300100

出　　处：《中华麻醉学杂志》2021年第9期1133-1137,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金面上项目(81772106);国家自然科学基金青年项目(82004076);国家自然科学基金青年项目(82002069)。

摘　　要：目的评价烟酰胺腺嘌呤二核苷酸(NAD^(+))介导的沉默信息调节因子1(SIRT1)去乙酰化活性在小鼠内毒素性急性肺损伤(ALI)中的作用。方法SPF级健康雄性C57BL/6小鼠25只,6~8周龄,体重20~25 g,野生(WT)型10只,NAD^(+)合成途关键酶烟酰胺单核苷酸腺苷酸转移酶1(NMNAT1)敲除(KO)型15只,采用随机数字表法,将WT型小鼠分为2组(n=5):对照组(WT+C组)和ALI组(WT+ALI组);将KO型小鼠分为3组(n=5):对照组(KO+C组)、ALI组(KO+ALI组)和ALI+NAD^(+)前体物质烟酰胺单核苷酸(NMN)组(KO+ALI+NMN组)。静脉注射LPS 15 mg/kg制备内毒素性ALI模型,KO+ALI+NMN组注射静脉注射LPS前1 h腹腔注射NMN 500 mg/kg。各对照组给予等容量生理盐水。注射LPS或生理盐水后12 h时,取腹主动脉血标本行血气分析,后处死小鼠留取肺组织,测定肺湿重/干重(W/D)比值,光镜下观察肺组织病理学改变,并行肺损伤评分;采用ELISA法检测肺组织IL-6、IL-1β、TNF-α含量,采用分光光度计法测定NAD^(+)含量,采用Western blot法测定肺组织SIRT1、乙酰化NF-κB(Ac-NF-κB)、乙酰化p53(Ac-p53)、乙酰化叉头框蛋白O1(Ac-FoxO1)、乙酰化过氧化物酶体增殖激活物受体γ辅激活因子(Ac-PGC1α)水平。结果与各C组比较,各ALI组pH值和PaO_(2)降低,PaCO_(2)、肺W/D比值、肺损伤评分、肺组织IL-6、IL-1β、TNF-α和NAD^(+)含量升高,SIRT1表达上调,Ac-NF-κB、Ac-p53、Ac-FoxO1和Ac-PGC1α表达下调(P<0.05)。与WT+ALI组比较,KO+ALI组pH值和PaO_(2)降低,PaCO_(2)、肺W/D比值、肺损伤评分、肺组织IL-6、IL-1β和TNF-α含量升高,NAD^(+)含量降低,SIRT1表达下调,Ac-NF-κB、Ac-p53、Ac-FoxO1和Ac-PGC1α表达上调(P<0.05)。与KO+ALI组比较,KO+ALI+NMN组pH值和PaO_(2)升高,PaCO_(2)、肺W/D比值、肺损伤评分、肺组织IL-6、IL-1β和TNF-α含量降低,NAD^(+)含量升高,SIRT1表达上调,Ac-NF-κB、Ac-p53、Ac-FoxO1和Ac-PGC1α表达下调(P<0.05)。结论NAD^(+)介导的SIRT1去乙酰化活性增强参与了小�Objective To evaluate the role of nicotinamide adenine dinucleotide(NAD^(+))-mediated deacetylation activity of silent information regulator 1(SIRT1)in endotoxin-induced acute lung injury(ALI)in mice.Methods Twenty-five SPF clean-grade healthy male C57BL/6 mice including 10 wild-type(WT)and 15 NMNAT1 conditional-knockout(KO)mice,aged 6-8 weeks,weighing 20-25 g,were selected.The WT mice were divided into 2 groups(n=5 each)using a random number table method:control group(group WT+C)and ALI group(group WT+ALI).The KO mice were divided into 3 groups(n=5 each)using a random number table method:control group(group KO+C),ALI group(group KO+ALI)and ALI plus NAD^(+) precursor substances nicotinamide mononucleotide(NMN)group(KO+LPS+NMN group).ALI was produced with lipopolysaccharide(LPS)15 mg/kg injected intravenously.NMN 500 mg/kg was intraperitoneally injected at 1 h before injection of LPS in KO+ALI+NMN group,while the equal volume of normal saline was given instead in control group.Blood samples were collected from the abdominal aorta at 12 h after LPS or normal saline injection for blood gas analysis,and the animals were then sacrificed and the lung tissues were removed for microscopic examination of pathologic changes which were scored and for determination of wet/dry weight ratio(W/D ratio),and interleukin-6(IL-6),IL-1βand tumor necrosis factor-alpha(TNF-α)contents(by enzyme-linked immunosorbent assay)and content of NAD^(+)(using a spectrophotometer)and levels of SIRT1,acetylated nuclear factor kappaB(Ac-NF-κB),acetylated p53(Ac-p53),acetylated FoxO1(Ac-FoxO1)and acetylated PGC1α(Ac-PGC1α)(by Western blot).Results Compared with group C,pH value and PaO_(2) were significantly decreased,the PaCO_(2),W/D ratio,lung injury score,contents of IL-6,IL-1β,TNF-αand NAD^(+)were increased,expression of SIRT1 was up-regulated,and expression of Ac-NF-κB,Ac-p53,Ac-FoxO1 and Ac-PGC1αwas down-regulated in group ALI(P<0.05).Compared with group WT+ALI,pH value and PaO_(2) were significantly decreased,the PaCO_(2),W/D ratio,l

关 键 词：内毒素血症 急性肺损伤 NAD 抗衰老酶1 去乙酰化 

分 类 号：R563[医药卫生—呼吸系统]