Role of dynamin-related protein 1 in hypoxia/reoxygenation-induced human renal tubular epithelial cell pyroptosis

陈雨; 侯卫平; 袁发焕

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Role of dynamin-related protein 1 in hypoxia/reoxygenation-induced human renal tubular epithelial cell pyroptosis

更新时间：2026-01-09

- Role of dynamin-related protein 1 in hypoxia/reoxygenation-induced human renal tubular epithelial cell pyroptosis
- Journal of Army Medical University Vol. 38, Issue 11, Pages: 1251-1256(2016)
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  CLC： R322.61;R329.28;R394.2
- Published：2016
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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].陆军军医大学学报(原第三军医大学学报),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 Amry Med Univ (J Third Mil Med Univ),2016,38(11):1251-1256.
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].陆军军医大学学报(原第三军医大学学报),2016,38(11):1251-1256. DOI：

　Chen Yu,Hou Weiping,Yuan Fahuan.Role of dynamin-related protein 1 in hypoxia/reoxygenation-induced human renal tubular epithelial cell pyroptosis[J].J Amry Med Univ (J Third Mil Med Univ),2016,38(11):1251-1256. DOI：

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% CO

和95%O

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（P0.05）；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（P0.05）。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% CO

). 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 (P0.05)

and the pyroptosis index were increased in the H/R group (P0.05). Mdivi-1 could dramatically decrease the level of ROS and pyroptosis and relieve the dysfunction of HK-2 cells (P0.05). Conclusion Drp1 can induce pyroptosis of human renal tubular epithelial cells through activating Nlrp3 in H/R injury.

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