敲除巨噬细胞低氧诱导因子-2α调控其炎性因子分泌对肾脏缺血再灌注损伤的保护作用  

Hypoxia inducible factor-2α knockout macrophages alleviate the renal ischemia-reperfusion injury via regulating the expression of inflammatory cytokines

作　　者：王健富[1] 刘东[1] 周结学[1] 蒙善东[1] 申升[1] 马娟[1] 邓庚国 朱春丽[1] 李琴[1] Wang Jianfu;Liu Dong;Zhou Jiexue;Meng Shandong;Shen Sheng;Ma Juan;Deng Gengguo;Zhu Chunli;Li Qin(Department of Organ Transplantation,Guangdong Second People’s Hospital,Guangzhou 510317,China)

机构地区：[1]广东省第二人民医院器官移植科,广州510317

出　　处：《中华实验外科杂志》2019年第10期1822-1825,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探讨敲除巨噬细胞中低氧诱导因子(HIF)-2α通过调控其炎症因子分泌对肾脏缺血再灌注损伤的保护作用.方法将HIF-2α^+/+和HIF-2α-/-小鼠随机分为假手术组(Sham组,n=20)和实验组(IR组,n=20),观察记录小鼠生存时间,再灌注24h处死小鼠获取相应标本,检测血清中肌酐(Cr)、尿素氮(BUN)及血清和肾脏中炎性因子白细胞介素(IL)-1β、IL-6、IL-10、肿瘤坏死因子-α(TNF-α)、干扰素-γ(IFN-γ)的表达水平;采用苏木精-伊红(HE)染色评估肾组织形态学改变.将HIF-2α^+/+和HIF-2α^-/-小鼠骨髓来源巨噬细胞分为常氧组和低氧组,酶联免疫吸附试验(ELISA)检测细胞上清中IL-1β、IL-6、IL-10、TNF-α及IFN-y水平.结果与HIF-2α^+/++IR组比较,HIF-2α^-/-+IR组小鼠生存时间显著延长(P<0.05),肾组织结构损伤明显减轻,血清Cr、BUN水平显著降低(t=5.750、6.506,P<0.01),肾组织中TNF-α、IFN-γ水平显著降低(t=10.849、4.716,P<0.01)而IL-10水平显著升高(t=-4.383,P< 0.05).HIF-2α^-/-+ IR组与HIF-2α^+/++ IR组IL-6、IL-1β水平差异无统计学意义(t=1.596、1.474,P>0.05).与HIF-2α^+/++低氧组比较,HIF-2α-/-+低氧组细胞上清中TNF-α、IL-6值显著降低(t=12.196、8.498,P<0.01)而IL-10值显著升高(t=-7.418,P<0.01).HIF-2α^+/++低氧组与HIF-2α^-/-+低氧组细胞上清中IFN-γ、IL-1β水平差异无统计学意义(t=-0.141、0.315,P>0.05).结论巨噬细胞中特异性敲除HIF-2α可减少其促炎因子分泌且增加抑炎因子分泌,从而减轻肾脏缺血再灌注损伤.Objective To investigate the protective effect of hypoxia inducible factor-2α(HIF-2α) knockout macrophages to renal ischemia-reperfusion injury through regulating the expression of inflammatory cytokines. Methods The HIF-2α^+/+ and HIF-2α^-/-mice were randomly divided into the sham operation group (n=20) and renal IR group (n=20). The survival time of mice was observed. Mice were sacrificed 24 h after renal reperfusion to obtain blood and renal samples. The expressions of blood urea nitrogen (BUN), creatinine (Cr) in serum and the inflammatory cytokines encoding interleukin (IL)-1β, IL-6, IL-10, tumor necrosis factor-α(TNF-α) and interferon-γ(IFN-γ) in serum as well as renal tissues were measured. In addition, the histomorphological changes were analyzed via hematoxylin and eosin (HE) staining. Macrophages derived from bone marrow of HIF-2α^+/+ and HIF-2α^-/- mice were divided into normoxic group and hypoxic group. The levels in cell culture supernatants of IL-1β, IL-6, IL-10, TNF-α and IFN-γ were examined by enzyme linked immunosorbent assay (ELISA). Results Compared with HIF-2α^+/++ IR group, mice in HIF-2α^-/-+ IR group had significantly longer survival time (P<0.05) and milder renal histopathologic injury. The serum levels of BUN and Cr in HIF-2α^-/-+ IR group were significantly lower than those in HIF-2α^+/++ IR group (t=5.750, 6.506, P<0.01). Relative to HIF-2α^+/++ IR group, the renal tissue expressions of TNF-α and IFN-γ in HIF-2α^-/-+ IR group were significantly reduced (t=10.849, 4.716, P<0.01) while IL-10 were dramatically increased (t=-4.383, P<0.05). There were no statistical significance of IFN-γ and L-1β expressions between HIF-2α^+/++ IR and HIF-2α^-/-+ IR group (t=-0.141, 0.315, P>0.05). Compared with HIF-2α^+/++ hypoxic group, bone marrow-derived macrophages (BMDMs) obtained from HIF-2α^-/-mice under hypoxic had significantly higher levels of TNF-α and IL-6 (t=12.196, 8.498, P<0.01) and lower levels of L-10 (t=-7.418, P<0.01) in cell culture supernatants. No signific

关 键 词：低氧诱导因子-2α 巨噬细胞 肾脏 缺血再灌注损伤 

分 类 号：R699.2[医药卫生—泌尿科学]