摘要
目的研究载脂蛋白A-I(ApoA-I)模拟肽D-4F对脂多糖(LPS)诱导的急性肺损伤(ALI)的保护作用及其机制。方法将6～8周龄的雌性BALB/C小鼠随机分为假手术(Sham)组、D-4F组、LPS组和LPS+D-4F组,每组6只。LPS组和LPS+D-4F组小鼠在气管内缓慢注射50μL LPS(1mg/kg)诱导ALI,Sham组和D-4F组小鼠则在气管内注射等量无菌0.9%氯化钠溶液;术后1h,D-4F组和LPS+D-4F组小鼠分别予腹腔内注射200μL D-4F(14mg/kg),Sham组和LPS组小鼠分别予腹腔内注射等量无菌0.9%氯化钠溶液。造模后6h,取小鼠肺泡灌洗液(BALF)和肺组织。采用二喹啉甲酸(BCA)法测定BALF总蛋白质质量浓度,根据肺组织湿/干质量比(W/D)评估肺水肿程度,经H-E染色观察肺组织病理学改变,采用ELISA法检测BALF中TNF-α、IL-6和IL-1β的表达水平,采用Western印迹法检测肺组织中磷酸化NF-κB(p-NF-κB)、磷酸化NF-κB抑制蛋白α(p-IκBα)、磷酸化NF-κB抑制蛋白激酶(p-IKK)的表达水平。结果 D-4F组与Sham组间BALF总蛋白质质量浓度、BALF炎性细胞因子表达水平、病理学评分,以及肺组织p-IKK、p-IκBα、p-NF-κB相对蛋白质表达水平的差异均无统计学意义(P值均>0.05);LPS组的BALF总蛋白质质量浓度、BALF炎性细胞因子表达水平、病理学评分,以及肺组织p-IKK、p-IκBα、p-NF-κB相对蛋白质表达水平均显著高于Sham组、D-4F组和LPS+D-4F组(P值分别<0.01、0.05)。LPS组小鼠肺组织W/D较Sham组有升高趋势,LPS+D-4F组的肺组织W/D较LPS组有降低趋势,但各组间肺组织W/D的差异均无统计学意义(P值均>0.05)。光学显微镜下可见,Sham组和D-4F组小鼠肺组织无明显损伤,LPS组小鼠的肺组织损伤明显,而LPS+D-4F组小鼠的肺组织损伤较LPS组显著减轻。结论 ApoA-I模拟肽D-4F通过抑制NF-κB通路减少BALF炎性细胞因子表达,减轻肺组织损伤,对小鼠ALI具有保护作用。
Objective To reveal the protective effect of apolipoprotein A-I(ApoA-I)mimetic peptide D-4F on lipopolysaccharide(LPS)-induced acute lung injury(ALI)and its possible mechanism.Methods FemaleBALB/C mice aged 6-8 weeks were randomly divided into four groups(n=6):sham group,D-4 Fgroup,LPSgroup and LPS+D-4 Fgroup.The model of ALI was established by intratracheal injection of LPS (1mg/kg,50μL).Mice received D-4F (14 mg/kg)or equal amount of normal saline (NS)intraperitoneally 1h afte rintratracheal instillation.The bronchoalveolar lavage fluid(BALF)and lung tissues were harvested at 6h after intratracheal instillation.The total protein concentration of BALF was determined by bicinchoninic acid(BCA).Thewet/dry ratio(W/D)was used to assess pulmonary edema.Pathological changes of lung tissues were observed byhematoxylin-eosin(H&E)staining.Moreover,the expression of tumor necrosis factor alpha(TNF-α),interleukin6(IL-6)and interleukin 1β(IL-1β)in BALF were measured by enzyme linked immunosorbent assay(ELISA),and phosphorylated nuclear factorκB(p-NF-κB),phosphorylated inhibitor of NF-κB alpha(p-IκBα)and phosphorylated inhibitor of NF-κB kinase(p-IKK)expression in lung tissues were detected by Western blotting.Results Therewas no significant difference in total protein concentration in BALF,inflammatory factor(TNF-α,IL-6and IL-1β)expression in BALF,pathological score,relative protein expression of p-IKK,p-IκBαor p-NF-κB between D-4Fgroup and sham group(all P>0.05).The parameters mentioned above were significantly higher in LPS groupthan those in sham group,D-4Fgroup and LPS+D-4Fgroup(P<0.01,0.05).Additionally,the W/D ratio inLPS group was higher than that in sham group,and the W/D ratio in LPS+D-4F group was lower than that in LPSgroup,but there was no significant difference in W/D ratio between the four groups(all P>0.05).Under lightmicroscope,lung tissues in sham group and D-4F group exhibited normal alveolar structure,but lung tissues in LPS group were significantly damaged.Compared with LPS group,lung tiss
作者
熊晨君
张越
李银娇
罗艳
XIONG Chenjun;ZHANG Yue;LI Yinjiao;LUO Yan(Department of Anesthesiology,Ruijin Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 200025,China)
出处
《上海医学》
CAS
北大核心
2018年第11期676-680,共5页
Shanghai Medical Journal
基金
上海市科学技术委员会基金(16ZR1421100)
关键词
炎症
脂多糖类
核因子ΚB
急性肺损伤
载脂蛋白A-I模拟肽
器官保护
Inflammation
Lipopolysaccharides
Nuclear factor-kappa B
Acute lung injury
Apolipoproteinmimetic peptide
Organ protection
