初级纤毛在肺微血管内皮细胞抗缺氧损伤中的作用研究  

Effect of Primary Cilia in Anti-hypoxia Damage of Pulmonary Microvascular Endothelial Cells

作　　者：秦荣[1] 李雪雁[1] 李凯 田永辉 刘天珍 崔冰冰 邵佳乐 陈克明 马慧萍 QIN Rong;LI Xue-yan;LI Kai;TIAN Yong-hui;LIU Tian-zhen;CUI Bing-bing;SHAO Jia-le;CHEN Ke-ming;MA Hui-ping(Collage of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China;Institute of Orthopaedics,940 Hospital of PLA Joint Logistics Support Forces, Lanzhou 730050, China;Department of Pharmacy,940 Hospital of PLA Joint Logistics Support Forces, Lanzhou 730050, China)

机构地区：[1]兰州理工大学生命科学与工程学院,兰州730050 [2]中国人民解放军联勤保障部队第九四〇医院(原解放军兰州总医院)骨科研究所,兰州730050 [3]中国人民解放军联勤保障部队第九四〇医院(原解放军兰州总医院)药剂科,兰州730050

出　　处：《解放军医药杂志》2019年第5期9-14,共6页Medical & Pharmaceutical Journal of Chinese People’s Liberation Army

基　　金：国家自然科学基金面上项目(81571847)

摘　　要：目的研究初级纤毛在肺内皮细胞抗缺氧损伤中的作用及机制。方法将肺微血管内皮细胞(PMVEC)培养至接近融合时,置于密闭缺氧盒中处理0、6、12、18和24h,观察处理不同时间后的细胞形态和初级纤毛长度的变化。用RNA干扰法(siRNA)抑制IFT-88的表达以观察PMVEC初级纤毛的发生情况,同时以无意义干扰序列作为阴性对照组(对照组)。将对照组和siRNA组同时置于缺氧环境0、6、12、18和24 h,检测各时间点的活性氧(ROS)和丙二醛(MDA)含量,谷胱甘肽氧化物酶(GSP-PX)和超氧化物歧化酶(SOD)活性。结果缺氧导致PMVEC细胞受损,且初级纤毛长度较常氧环境下显著增加(P<0.01)。IFT88表达被干扰后PMVEC的初级纤毛发生受抑制。对照组PMVEC的ROS、MDA水平在缺氧后逐渐升高,12 h时达到峰值,之后开始下降,24 h后又略有回升;GSH-PX和SOD活性逐渐升高,至12 h时达到峰值,之后持续下降(P<0.01)。siRNA组的ROS和MDA水平随着缺氧时间的延长持续升高,GSH-PX和SOD活性无明显变化。结论纤毛参与了胞外缺氧及氧化胁迫信息向胞内的传递并在抗缺氧损伤中发挥了不可或缺的作用。Objective To investigate effect and its mechanism of primary cilia in anti-hypoxia damage of pulmonary microvascular endothelial cells (PMVEC). Methods PMVECs were cultivated to near fusion, and then the cells were placed to airtight hypoxia-box in 0 h, 6 h, 12 h, 18 h and 24 h of treatment respectively, and changes of cell morphological and length of primary cilia were observed at different times. RNA interference (siRNA) was used to inhibit ITF-88 expression in order to observe occurrence of PMVEC primary cilia, and nonsense RNA sequence was used as negative control (NC) group. Both siRNA and NC groups were simultaneously placed into hypoxia environment for 0 h, 6 h, 12 h, 18 h and 24 h, and then contents of reactive oxygen species (ROS) and malondialdehyde (MDA) and activities of glutathion peroxidase (GSH-PX) and superoxide dismutase (SOD) were detected at different times. Results PMVEC cells were damaged and length of primary cilia was significantly increased under hypoxia environment compared with those under normoxia environment ( P <0.01). Occurrence of PMVEC primary cilia was inhibited after interfering ITF-88 expression. In NC group, PMVEC intracellular levels of ROS and MDA were gradually increased under hypoxia environment, and peaked after hypoxia for 12 h, and then the levels began to decrease, and then the levels were increased a little after hypoxia for 24 h;activities of GSH-PX and SOD were gradually increased and peaked after hypoxia for 12 h, and then the activities were decrease continuously ( P <0.01). In siRNA group, ROS and MDA levels were increased continuously with prolonging hypoxia time ( P <0.01), and no activities of SOD and GSH-PX were found. Conclusion Primary cilia participates transmitting of anoxia out of cells and oxidative stress information into cells, and it plays an indispensable role in anti-hypoxia damage.

关 键 词：缺氧 肺微血管内皮细胞 初级纤毛 RNA干扰 抗氧化酶 活性氧 

分 类 号：R594.3[医药卫生—内科学] Q26[医药卫生—临床医学]