HIF-1α在氢抑制脂多糖诱导小鼠巨噬细胞炎症反应中的作用  

Role of HIF-1αin hydrogen-induced inhibition of lipopolysaccharide-induced inflammatory responses in mouse macrophages

作　　者：王慧星[1] 韩晨阳 陈红光[2] 刘靖芷[1] 马文庭[1] 李全波[1] 蒋宁[3] 霍小东[3] 于泳浩[2] 史可梅[1] Wang Huixing;Han Chenyang;Chen Hongguang;Liu Jingzhi;Ma Wenting;Li Quanbo;Jiang Ning;Huo Xiaodong;Yu Yonghao;Shi Kemei(Pain Management Center,Second Hospital of Tianjin Medical University,Tianjin 300211,China;Department of Anesthesiology,General Hospital of Tianjin Medical University,Tianjin Institute of Anesthesiology,Tianjin 300200,China;Tianjin Medical University Central Laboratory of Second Hospital,Tianjin 300211,China)

机构地区：[1]天津医科大学第二医院疼痛治疗中心,300211 [2]天津医科大学总医院麻醉科,天津市麻醉学研究所,300200 [3]天津医科大学第二医院中心实验室,300211

出　　处：《中华麻醉学杂志》2020年第7期881-884,共4页Chinese Journal of Anesthesiology

基　　金：天津市自然科学基金项目(18JCQNJC13100)。

摘　　要：目的评价缺氧诱导因子-1α(HIF-1α)在氢抑制脂多糖(LPS)诱导小鼠巨噬细胞炎症反应中的作用。方法体外培养小鼠RAW264.7巨噬细胞,按照随机数字表法分为4组(n=24):对照组(C组)、LPS组(L组)、富氢液+LPS组(H+L组)和富氢液+LPS+HIF-1α抑制剂二甲氧基雌二醇(2ME2)组(H+L+M组)。L组加入LPS 1μg/ml孵育6 h;L+H组先加LPS,换培养基为0.6 mmol/L富氢液共孵育6 h;H+L+M组先给予2ME210μmol/L孵育30 min,然后加入LPS和富氢液孵育6 h。采用Western blot法测定HIF-1α、Beclin-l、BNIP3和LC3的表达;采用ELISA法检测上清液TNF-α、IL-6和IL-1β的浓度,采用透视电镜观察细胞自噬小体数量。结果与C组比较,L组上清液TNF-α、IL-6和IL-1β浓度升高,巨噬细胞HIF-1α、Beclinl和BNIP3表达上调,LC3Ⅱ/LC3Ⅰ比值升高,自噬小体数量增多(P<0.05);与L组比较,H+L组上清液TNF-α、IL-6和IL-1β浓度降低,巨噬细胞HIF-1α、Beclin-l、BNIP3表达上调、LC3Ⅱ/LC3Ⅰ比值增高,自噬小体数量增多(P<0.05);与H+L组比较,H+L+M组上清液TNF-α、IL-6和IL-1β的浓度降低,巨噬细胞HIF-1α、Beclin-l和BNIP3表达下调,LC3Ⅱ/LC3Ⅰ比值降低,自噬小体数量减少(P<0.05)。结论HIF-1α介导细胞自噬激活参与了氢抑制LPS诱导小鼠巨噬细胞炎症反应的过程。Objective To evaluate the role of hypoxia-inducible factor-1α(HIF-1α)in hydrogen-induced inhibition of lipopolysaccharide(LPS)-induced inflammatory responses in mouse macrophages.Methods The mouse RAW264.7 macrophages cultured in vitro were divided into 4 groups(n=24 each)according to the random number table method:control group(C group),LPS group(L group),hydrogen-rich solution plus LPS group(H+L group),and hydrogen-rich solution plus LPS plus HIF-1αinhibitor 2-methoxyestradiol(2ME2)group(H+L+M group).LPS 1μg/ml was added,and the cells were incubated for 6 h in group L.In group L+H,LPS was added first,the medium was changed to 0.6 mmol/L hydrogen-rich solution,and cells were incubated for 6 h.In group H+L+M,2ME210μmol/L was given first,cells were then incubated for 30 min,LPS and hydrogen-rich solution were added,and cells were incubated for 6 h.Western blot was used to determine the expression of HIF-1α,Beclin-1,Bcl-2/E1B-19 kDa interacting protein 3(BNIP3)and LC3.Enzyme-linked immunosorbent assay was used to detect the concentrations of tumor necrosis factor(TNF)-α,interleukin(IL)-6,and IL-1βin the supernatant.The number of autophagosomes was observed using a transmission electron microscope.Results Compared with group C,the concentrations of TNF-α,IL-6 and IL-1βin the supernatant were significantly increased,the expression of HIF-1α,Beclinl and BNIP3 in macrophages was up-regulated,the ratio of LC3Ⅱ/LC3Ⅰwas increased,and the number of autophagosomes was increased in group L(P<0.05).Compared with group L,the concentrations of TNF-α,IL-6 and IL-1βwere significantly decreased,the expression of HIF-1α,Beclin-1 and BNIP3 in macrophages was up-regulated,LC3Ⅱ/LC3Ⅰratio was increased,and the number of autophagosomes was increased in group H+L(P<0.05).Compared with group H+L,the concentrations of TNF-α,IL-6 and IL-1βin the supernatant were significantly decreased,the expression of HIF-1α,Beclin-1,and BNIP3 in macrophages was down-regulated,and the ratio of LC3Ⅱ/LC3Ⅰwas decreased,and the nu

关 键 词：缺氧诱导因子1 Α亚基 氢 脂多糖类 巨噬细胞 炎症 

分 类 号：R364.5[医药卫生—病理学]