肾小管上皮细胞G2-M期阻滞在缺氧诱导的肾间质纤维化中的作用  

Effect of G2/M phase arrest in renal tubular epithelial cells on hypoxia-induced renal interstitial fibrosis

作　　者：刘婷[1] 党杨杰 刘利敏[1] 张磊[1] 付爱萍[1] 张玉明[1] 吴笛[3] 杜锐 孙世仁[1] LIU Ting;DANGYang-jie;LIU Li-min;ZHANG Le;FU Aiping;ZHANG Yu-ming;WU Di;DU Rui;SUN Shi-ren(State Key Laboratory of Cancer Biology, Department of Nephrology ,Xijing Hospital, Fourth Military Medical University,Xi’an 710032, China;State Key Laboratory of Military Stomatology VNational Clinical Research Center for Oral DiseasesPShaanxi Engineering Research Center for Dental Materials and Advancee Manufacture,Deeartment ooAnesthesiology,School 〇oStomatology, The Fourtt Militarr Medical University,Xi'an 710032,China;Cadet Brigade,Fourth Military Medical University,Xi'an 710032, China;Deecirtment of Radiation Oncology,Navv General Hossital’Beijing,100048,China)

机构地区：[1]第四军医大学西京医院肾内科,国家肿瘤重点实验室,陕西西安710032 [2]军事口腔医学国家重点实验室,口腔疾病国家临床医学研究中心,陕西省口腔生物工程技术研究中心,第四军医大学口腔医学院麻醉科,陕西西安710032 [3]第四军医大学学员一旅 [4]海军总医院肿瘤诊疗中心,北京100048

出　　处：《东南大学学报（医学版）》2017年第2期240-245,共6页Journal of Southeast University(Medical Science Edition)

基　　金：国家自然科学基金面上项目(NO.81370789)

摘　　要：目的:探讨肾小管上皮细胞G2-M期阻滞在缺氧诱导的肾间质纤维化中的作用。方法:体外实验,人肾小管上皮细胞(HK2)分别置于常氧21%和缺氧1%的孵箱里,培养24,48 hrs。采用流式细胞术检测缺氧48 hrs后上皮细胞的周期分布;采用Western blot法检测Collagen4A1(COL4A1)及α-平滑肌肌动蛋白(α-SMA)的蛋白表达水平。单侧输尿管梗阻(UUO)14 d小鼠模型,HE及Masson染色观察肾组织病理改变及纤维化程度,免疫组化染色观察α-SMA的表达及部位;Western blot检测低氧标记分子HIF-1α、细胞周期蛋白cyclinB1和cyclinD1的蛋白水平。结果:与常氧组比较,缺氧组肾小管上皮细胞G2/M期比例增高(P<0.05),同时α-SMA和COL4A1的蛋白表达水平增高(P<0.05);与假手术组比较,HE及Masson染色显示模型组肾组织纤维化程度增高,α-SMA的表达增高且主要分布在间质中;Western blot结果显示:HIF-1α及G2/M期标记物cyclinB1/cyclinD1比值增加。结论:肾小管上皮细胞G2-M期阻滞参与缺氧诱导的肾间质纤维化。O bjective:To investigate effect of G2/M phase arrest in renal tubular epithelial cells on hypoxiainducedrenal interstitial fibrosis.M eth od s:V t o,HK2cells were subjected to hypoxia(1%〇2)or normoxia(21%〇2)for24and48hrs.We examined the cell cycle distribution of HK2cells by flow^cytometry,of C0L4A1and a^SMA was detected by Western blot48hrs after hypoxia$We observed the and renal tissue filDrosis degree by HE and Masson staining.The expression of a"SMA was detected staining in U U0model.Then we tested the protein level of HIF_1a,cyclinB1and cyclinD1by W esternblot in U U0model.R esu lts:Compared with normoxia group,hypoxia group showed a significant increase inthe percentage of G2/M stage in HK2cells(P<0.05).Hypoxia induced a significant increase of a-SMA andC0L4A1in protein lev el(P<0.05)$Compared witii the sham group,HE and Masson staining revealed the increaseddegree of renal tissue filDrosis and increased expression of a"SMA that was HIF-1a and G2/M phase marker cyclinB1/cyclinD1was increased in U U0model by Western blot.C onclusion:Chronic hypoxia induced renal interstitial fibrosis is associated with G2/M arrest in renal tubular

关 键 词：缺氧 G2-M阻滞 肾小管上皮细胞 肾间质纤维化 

分 类 号：R692.2[医药卫生—泌尿科学]