Elabela活性片段ELA14在对比剂致大鼠肾小管上皮细胞凋亡中的作用机制  

The role and mechanism of Elabela (ELA) active fragment ELA14 in the apoptosis of rat renal tubular epithelial cells induced by iopromide

作　　者：孟丹心 李益民 美合日阿依·约麦尔 陆治平 黄进 MENG Dan-xin;LI Yi-min;Meiheriayi·Yuemaier;LU Zhi-ping;HUANG Jin(Department of Cardiovascular,The Affiliated Brain Hospital of Nanjing Medical University,Nanjing 210029,Jian-gsu,China;Department of Cardiovascular,The Affiliated Sir Run Run Hospital of Nanjing Medical University,Nan-jing 211166,Jiangsu,China)

机构地区：[1]南京医科大学附属脑科医院心血管内科,南京210029 [2]南京医科大学附属逸夫医院心血管内科,南京211166

出　　处：《医学研究生学报》2022年第8期841-846,共6页Journal of Medical Postgraduates

基　　金：南京市医学重点科技发展项目(ZKX16066)。

摘　　要：目的 Elabela活性片段ELA32在对比剂肾病中可减轻肾损伤,但ELA14在对比剂肾病中作用仍不明确。文章探究Elabela活性片段ELA14在对比剂致大鼠肾小管上皮细胞(NRK-52E细胞)凋亡中作用及机制。方法 实验分组:对照组、碘普罗胺(IOP)组(加入21 mgI/mL IOP)、IOP+高、低剂量ELA32组(21 mgI/mL IOP与30 nmol/L、3μmol/L ELA32共同孵育)、IOP+高、低剂量ELA14组(21 mgI/mL IOP与30 nmol/L、3μmol/L ELA14共同孵育)。用流式细胞仪检测细胞凋亡率、CCK-8评估细胞活力,酶联免疫法测定炎症因子,线粒体活性氧法测定ROS强度,Western blot、qRT-PCR检测细胞mRNA和蛋白表达量。结果 IOP组细胞凋亡率[(17.306±0.848)%]较对照组[(4.860±0.182)%]明显升高(P<0.05),且ROS强度、炎性因子水平(IL-6、TNF-α、NRF2 mRNA及ICAM-1mRNA)及DNA损伤相关蛋白(caspase、CHK1、ATR)的表达均较对照组明显升高(P<0.05)。高、低剂量的ELA14[(8.813±0.397)%、(11.800±0.756)%]可明显改善IOP[(17.306±0.848)%]诱导的细胞凋亡率(P<0.05),但与同等剂量的ELA32效果差异无统计学意义(P>0.05),另外ELA32和ELA14均可改善ROS水平、炎症因子水平及DNA损伤相关蛋白的表达。IOP组细胞荧光强度(3 842.780±444.146)较对照组(240.18±80.941)明显升高,而IOP+高、低剂量ELA32组和IOP+高、低剂量ELA14组细胞荧光强度(807.810±98.798、2 008.460±332.571、1 362.716±415.551、2 325.856±455.404)较IOP组明显减弱(P<0.05)。结论 与ELA32相似,ELA14也可抑制IOP诱导的NRK-52E细胞氧化应激、炎症反应及细胞凋亡,并呈剂量依赖性,该作用可能通过抑制氧化炎症反应实现。Objective The active fragment of Elabela ELA32 can reduce renal injury in contrast-induced nephropathy, but the role of ELA14 in it remains unclear. This paper aims to explore the role and mechanism of Elabela active fragment ELA14 in the apoptosis of rat renal tubular epithelial cells(NRK-52 E cells) induced by contrast medium iopromide. Methods NRK-52 E cells were divided into 6 groups, control group, IOP group(21 mgI/mL IOP) and IOP+ high-dose and low-dose ELA32 group(21 mgI/mL IOP incubated with 30 nmol/L and 3 μmol/L ELA32), IOP+ high-dose and low-dose ELA14 groups(21 mgI/mL IOP incubated with 30 nmol/L and 3 μmol/L ELA14). After incubating cells, flow cytometer was used to detect apoptosis rate, CCK-8 to assess cell viability, enzyme-linked immunosorbent assay to detect inflammatory factors, mitochondrial reactive oxygen methods to detect ROS intensity, Western-blot and qRT-PCR to detect expression of mRNA and protein in cells. Results The apoptosis rate of IOP group [(17.306±0.848) %] was significantly higher than that of control group [(4.860±0.182) %](P<0.05). The levels of ROS, inflammatory factors(IL-6, TNF-α, NRF2 mRNA and ICAM-1 mRNA) and the expressions of DNA damage-related proteins(caspase, CHK1 and ATR) were significantly increased compared with the control group(P<0.05). High and low doses of ELA14[(8.813±0.397) %(11.800±0.756) %] could significantly improve the apoptosis rate induced by IOP[(17.306±0.848) %](P<0.05), but there was no significant difference with the same dose of ELA32(P>0.05). In addition, both ELA32 and ELA14 could improve ROS levels, inflammatory factors and the expression of DNA damage-related proteins. The fluorescence intensity of IOP group(3 842.780±444.146) was significantly higher than that of control group(240.18±80.941). The fluorescence intensity of IOP+ high-dose and low-dose ELA32 groups and IOP+ high-dose and low-dose ELA14 groups were(807.810±98.798, 2008.460±332.571, 1362.716±415.551, 2325.856±455.404) was significantly lower than that in IOP group(

关 键 词：Elabela 肾损伤 对比剂 氧化应激 

分 类 号：R692[医药卫生—泌尿科学]