[1]陈雨,侯卫平,袁发焕.动力相关蛋白-1在缺氧/复氧诱导人肾小管上皮细胞焦亡机制中的作用[J].第三军医大学学报,2016,38(11):1251-1256.
 Chen Yu,Hou Weiping,Yuan Fahuan.Role of dynamin-related protein 1 in hypoxia/reoxygenation-induced human renal tubular epithelial cell pyroptosis[J].J Third Mil Med Univ,2016,38(11):1251-1256.
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动力相关蛋白-1在缺氧/复氧诱导人肾小管上皮细胞焦亡机制中的作用(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
38卷
期数:
2016年第11期
页码:
1251-1256
栏目:
基础医学
出版日期:
2016-06-15

文章信息/Info

Title:
Role of dynamin-related protein 1 in hypoxia/reoxygenation-induced human renal tubular epithelial cell pyroptosis
作者:
陈雨侯卫平袁发焕
第三军医大学新桥医院肾内科,全军肾脏病中心,重庆市肾脏病研究所,国家中医药管理局重点专科
Author(s):
Chen Yu Hou Weiping Yuan Fahuan

Department of Nephrology, Kidney Disease Center, Chongqing Institute of Kidney Disease, Key Department of State Administration of Traditional Chinese Medicine, Xinqiao Hospital, Third Military Medical University, Chongqing, 400037, China

关键词:
动力相关蛋白1Nod样受体热蛋白结构域相关蛋白3细胞焦亡人肾小管上皮细胞缺氧/复氧损伤
Keywords:
dynamin-related protein 1 nucleotide binding oligomerization domain-like receptor family pyrin domain containing 3 pyroptosis human renal tubular epithelial cell hypoxia/reoxygenation injury
分类号:
R322.61; R329.28; R394.2
文献标志码:
A
摘要:

目的      观察人肾小管上皮细胞(HK-2细胞)缺氧/复氧(hypoxia/reoxygenation, H/R)损伤时线粒体动力相关蛋白1(dynamin-related protein 1, Drp1)、Nod样受体热蛋白结构域相关蛋白3(nucleotide binding oligomerization domain-like receptor family pyrin domain containing 3, Nlrp3)以及炎性因子的表达改变和细胞焦亡情况,探讨Drp1在H/R诱导肾小管上皮细胞焦亡中的作用。      方法      HK-2细胞用含10%胎牛血清的DMEM/F12液培养于37 ℃、5% CO2和95%O2的恒温培养箱。将细胞分为正常对照组、H/R组、H/R+Drp1抑制剂(H/R+Mdivi-1)组、H/R+siNlrp3腺病毒(H/R+siNlrp3)组、H/R+空腺病毒(H/R+Scra)组。检测细胞培养液上清乳酸脱氢酶LDH活性,Western blot检测Drp1、Nlrp3、凋亡相关蛋白-1(caspase-1)、IL-1β表达,2′,7′-二氯荧光黄双乙酸盐(DCFH-DA)检测细胞活性氧(ROS)水平,流式分析仪检测细胞焦亡情况。       结果      与正常对照组比较,H/R组培养液上清LDH和细胞内ROS水平明显升高,Drp1、Nlrp3、caspase-1、IL-1β表达增高,细胞焦亡明显增加,差异有统计学意义(P<0.05);与H/R组比较,H/R+Mdivi-1组培养液上清LDH和细胞内ROS水平明显降低,Nlrp3、caspase-1、IL-1β表达降低,细胞焦亡明显减少,差异有统计学意义(P<0.05)。       结论      在H/R条件下,Drp1通过激活Nlrp3炎性体诱导人肾小管上皮细胞焦亡。

Abstract:

Objective      To study the expressions of dynamin-related protein 1 (Drp1), nucleotide binding oligomerization domain-like receptor family pyrin domain containing 3 (Nlrp3) and inflammatory cytokines and the pyroptosis during hypoxia/reoxygenation (H/R) injury in human renal tubular epithelial cell line (HK-2), and discuss the relationship between Drp1 and H/R-induced pyroptosis of HK-2 cells.       Methods      HK-2 cells were cultured in DMEM/F12 containing 10% fetal bovine serum in a thermostatic incubator (37 ℃, 5% CO2). Then the cells were divided into normal control group, H/R group, H/R+Mdivi-1 (Drp1 inhibitor) group, H/R+siNlrp3 group and H/R+Scra group. LDH activity in the supernatant of cell culture was tested with an automatic biochemical analyzer. The expressions levels of Drp1, Nlrp3, caspase-1 and IL-1β were detected by Western blotting. Reactive oxygen species was measured by 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) assay. Pyroptosis was analyzed with a flow cytometer.       Results      The H/R group exhibited higher celluar ROS level and higher expressions of Drp1, Nlrp3, caspase-1 and IL-1β proteins than the normal control group (P<0.05), and the pyroptosis index were increased in the H/R group (P<0.05). Mdivi-1 could dramatically decrease the level of ROS and pyroptosis and relieve the dysfunction of HK-2 cells (P<0.05).       Conclusion      Drp1 can induce pyroptosis of human renal tubular epithelial cells through activating Nlrp3 in H/R injury.

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更新日期/Last Update: 2016-05-29