阿托伐他汀调控PERK/eIF2α/CHOP通路减轻造影剂诱导的大鼠肾小管上皮细胞凋亡

doi:10.3877/cma.j.issn.2095-3216.2019.03.002

目的

研究造影剂肾病大鼠肾脏中葡萄糖调节蛋白78(GRP78)、内质网调节激酶(PERK)、真核起始因子2α(eIF2α)及C/EBP同源蛋白质(CHOP)的表达情况，探讨内质网应激在造影剂肾病发病中的作用及阿托伐他汀的干预作用。

方法

60只大鼠随机分为4组：对照组、模型组和高、低剂量阿托伐他汀组(80 mg，40 mg)，每组15只。分别于注射造影剂后24、48、72 h留取血清；检测各组大鼠的血清尿素氮(BUN)、血清肌酐(Scr)；TUNEL法及Western印迹法测casepase-3的表达检测肾小管上皮细胞凋亡；免疫组化和Western印迹法检测各组大鼠肾组织GRP78、p-eIF2α、p-PERK及CHOP的表达。

结果

与对照组相比，模型组大鼠BUN、Scr显著升高，细胞凋亡严重，GRP78、p-eIF2α、p-PERK及CHOP的表达均显著升高(P< 0.05);与模型组相比，高、低剂量阿托伐他汀组，BUN、Scr显著下降，凋亡指数降低，GRP78、p-eIF2α、p-PERK及CHOP的表达显著下调，但仍高于对照组，差异均达到统计学意义(P<0.05)；高、低剂量阿托伐他汀组之间上述各指标差异均不显著。

结论

PERK/eIF2α/CHOP通路介导的内质网应激可能参与大鼠造影剂肾病的发生发展；阿托伐他汀在造影剂诱导的肾脏损伤中发挥保护作用，这可能与其调节PERK/eIF2α/CHOP通路，从而减轻内质网应激有关。

关键词: 造影剂, 内质网应激, 葡萄糖调节蛋白78, 阿托伐他汀

Objective

To investigate the expression of glucose regulated protein 78 (GRP78), protein kinase R-like ER kinase (PERK), eukaryotic initiation factor 2α (eIF2α), and C/EBP homologous protein (CHOP), in order to evaluate the effect of endoplasmic reticulum stress in rats with contrast induced-acute kidney injury (CI-AKI), and observe the possible protective effect of atorvastatin in CI-AKI rats.

Methods

Sixty Wistar rats were randomly divided into control group, CI-AKI model group, high-dose atorvastatin group (80 mg), and low-dose atorvastatin group (40 mg) with 15 rats each. At 24 h, 48 h, and 72 h after the rat model was established, BUN and Scr were detected. TUNEL and Western blotting were used to detect the apoptosis of renal tubular epithelial cells and caspase-3 expression, respectively. Immunohistochemistry and Western blotting were used to detect the expression of GRP78, p-eIF2α, p-PERK, and CHOP in the rat renal tissues.

Results

Compared with the control group, the model group had higher levels of BUN and Scr, more apoptotic cells, and more expression of GRP78, p-eIF2α, p-PERK, and CHOP (P<0.05). Both the high- and low-dose atorvastatin groups had lower levels of BUN and Scr, fewer apoptotic cells, and less expression of GRP78, p-eIF2α, p-PERK, and CHOP than the model group, which were yet higher than those of the control group (P<0.05). The differences of the levels of BUN, Scr, apoptosis of tubular cells, and expression of GRP78, p-eIF2α, p-PERK and CHOP proteins were not significant between the high-dose and low-dose atorvastatin groups.

Conclusion

Endoplasmic reticulum stress mediated by the PERK/eIF2α/CHOP pathway may be involved in the development of contrast- enhanced nephropathy in rats. Atorvastatin may play a protective role in contrast-induced renal injury by its regulation of the PERK/eIF2α/CHOP pathway leading to reduced endoplasmic reticulum stress.

Key words: Contrast medium, Endoplasmic reticulum stress, Glucose-regulated protein 78, Atorvastatin

图1 大鼠肾组织光镜(HE 400×)及电镜(5 000×)图片

表1 造模后不同时间点各组大鼠BUN和Scr水平(±s, n＝15)

时间	血肌酐(μmol/L)				血尿素氮(mmol/L)
时间	对照组	模型组	高剂量阿托伐他汀组	低剂量阿托伐他汀组	对照组	模型组	高剂量阿托伐他汀组	低剂量阿托伐他汀组
术前	34.6±2.9	35.5±3.2	33.9±2.1	34.8±2.4	7.4±4.5	7.2±3.6	7.2±3.5	7.6±4.5
术后24 h	36.9±1.9	67.8±5.9^a	47.9±6.0^ab	48.7±4.2^ab	7.6±3.7	35.4±3.9^a	17.8±4.8^ab	18.1±3.9^ab
术后48 h	36.8±3.2	78.8±6.4^a	57.8±5.7^ab	59.6±6.1^ab	7.3±5.5	35.6±5.1^a	18.5±5.0^ab	18.3±5.7^ab
术后72 h	35.6±2.8	76.2±3.8^a	55.3±3.9^ab	57.1±2.1^ab	7.8±3.2	36.1±4.6^a	17.8±5.3^ab	18.2±4.4^ab

表1 造模后不同时间点各组大鼠BUN和Scr水平(±s, n＝15)

时间	血肌酐(μmol/L)				血尿素氮(mmol/L)
时间	对照组	模型组	高剂量阿托伐他汀组	低剂量阿托伐他汀组	对照组	模型组	高剂量阿托伐他汀组	低剂量阿托伐他汀组
术前	34.6±2.9	35.5±3.2	33.9±2.1	34.8±2.4	7.4±4.5	7.2±3.6	7.2±3.5	7.6±4.5
术后24 h	36.9±1.9	67.8±5.9^a	47.9±6.0^ab	48.7±4.2^ab	7.6±3.7	35.4±3.9^a	17.8±4.8^ab	18.1±3.9^ab
术后48 h	36.8±3.2	78.8±6.4^a	57.8±5.7^ab	59.6±6.1^ab	7.3±5.5	35.6±5.1^a	18.5±5.0^ab	18.3±5.7^ab
术后72 h	35.6±2.8	76.2±3.8^a	55.3±3.9^ab	57.1±2.1^ab	7.8±3.2	36.1±4.6^a	17.8±5.3^ab	18.2±4.4^ab

图2 造模48 h时各组大鼠肾脏细胞凋亡情况(TUNEL 400×)

图3 各组大鼠Caspase－3活性蛋白质表达(Western印迹)

图4 各组大鼠肾组织GRP78、p－eIF2α、p－PERK及CHOP蛋白的表达(Western印迹)

图5 各组大鼠肾组织GRP78、p－eIF2α、p－PERK及CHOP的表达(免疫组化400×)

图6 各组大鼠肾组织细胞色素c和Fas的蛋白表达(Western印迹)

[1]	Nash K, Hafeez A, Hou S. Hospital-acquired renal insufficiency [J]. Am J Kidney Dis, 2002, 39(5): 930-936.
[2]	Peng PA, Wang L, Ma Q, et al. Valsartan protects HK-2 cells from contrast media-induced apoptosis by inhibiting endoplasmic reticulum stress [J]. Cell Biol Int, 2015, 39(12): 1408-1417.
[3]	Wu CT, Sheu ML, Tsai KS, et al. The role of endoplasmic reticulum stress-related unfolded protein response in the radiocontrast medium-induced renal tubular cell injury [J]. Toxicol Sci, 2010, 114(2): 295-301.
[4]	Han Y, Zhu G, Han L, et al. Short-term rosuvastatin therapy for prevention of contrast-induced acute kidney injury in patients with diabetes and chronic kidney disease [J]. J Am Coll Cardiol, 2014, 63(1): 62-70.
[5]	Leoncini M, Toso A, Maioli M, et al. Early high-dose rosuvastatin for contrast-induced nephropathy prevention in acute coronary syndrome: Results from the PRATO-ACS Study [J]. J Am Coll Cardiol, 2014, 63(1): 71-79.
[6]	Su X, Xie X, Liu L, et al. Comparative effectiveness of 12 treatment strategies for preventing contrast-induced acute kidney injury: a systematic review and Bayesian network meta-analysis [J]. Am J Kidney Dis, 2017, 69(1): 69-77.
[7]	Quintavalle C, Fiore D, De Micco F, et al. Impact of a high loading dose of atorvastatin on contrast-induced acute kidney injury [J]. Circulation, 2012, 126(25): 3008-3016.
[8]	Breder I, Coope A, Arruda AP, et al. Reduction of endoplasmic reticulum stress-a novel mechanism of action of statins in the protection against atherosclerosis [J]. Atherosclerosis, 2010, 212(1): 30-31.
[9]	Wu CT, Weng TI, Chen LP, et al. Involvement of caspase-12-dependent apoptotic pathway in ionic radiocontrast urografin-induced renal tubular cell injury [J]. Toxicol Appl Pharmacol, 2013, 266(1): 167-175.
[10]	Giacoppo D, Madhavan MV, Baber U, et al. Impact of contrast-induced acute kidney injury after percutaneous coronary intervention on short- and long-term outcomes: pooled analysis from the HORIZONS-AMI and ACUITY trials [J]. Circ Cardiovasc Interv, 2015, 8(8): e002475.
[11]	Michael A, Faga T, Pisani A, et al. Molecular mechanisms of renal cellular nephrotoxicity due to radiocontrast media [J]. Biomed Res Int, 2014, 2014: 249810.
[12]	Andreucci M, Lucisano G, Faga T, et al. Differential activation of signaling pathways involved in cell death, survival and inflammation by radiocontrast media in human renal proximal tubular cells [J]. Toxicol Sci, 2011, 119(2): 408-416.
[13]	袁杨刚，丁桂霞，张爱华. 内质网应激与肾脏损伤[J]. 中华肾脏病杂志，2011, 27(12): 949-952.
[14]	Zhang K, Kaufman RJ. From endoplasmic reticulum stress to the inflammatory response [J]. Nature, 2008, 454(7203): 455-462.
[15]	Lu TH, Tseng TJ, Su CC, et al. Arsenic induces reactive oxygen species-caused neuronal cell apoptosis through JNK/ERK- mediated mitochondria-dependent and GRP78/CHOP- regulated pathways [J]. Toxicol Lett, 2014, 224(1): 130-140.
[16]	Xinwei J, Xianghua F, Jing Z, et al. Comparison of usefulness of simvastatin 20 mg versus 80 mg in preventing contrast-induced nephropathy in patients with acute coronary syndrome undergoing percutaneous coronary intervention [J]. Am J Cardiol, 2009, 104(4): 519-524.

[1]	宋勇, 李东炫, 王翔, 李锐. 基于数据挖掘法分析3 种超声造影剂不良反应信号[J/OL]. 中华医学超声杂志（电子版）, 2024, 21(09): 890-898.
[2]	覃斯, 刘一铭, 周静雯, 李昀芸, 刘广健. Sonazoid^®超声造影不同参数成像效果的体外研究[J/OL]. 中华医学超声杂志（电子版）, 2023, 20(12): 1287-1293.
[3]	赵鑫, 郝磊, 朱丽静, 土继政, 王博娟, 张凯, 王兴华. 超声造影评价肺腺癌与肺鳞癌血流灌注特征的价值研究[J/OL]. 中华医学超声杂志（电子版）, 2023, 20(11): 1181-1185.
[4]	蒋清凌, 覃斯, 胡美玉, 谢佩怡, 刘广健. 超声检查对肠系膜上动脉综合征的诊断价值[J/OL]. 中华医学超声杂志（电子版）, 2023, 20(11): 1174-1180.
[5]	董湘如, 初银珠, 黎富新, 张锋, 张玉莹, 吴长君. 超声造影与多层增强螺旋CT对肾占位性病变诊断价值的对比研究[J/OL]. 中华医学超声杂志（电子版）, 2022, 19(11): 1238-1243.
[6]	孙鸿坤, 艾虹, 陈正. 内质网应激介导的牙周炎骨改建失衡的研究进展[J/OL]. 中华口腔医学研究杂志(电子版), 2024, 18(04): 211-218.
[7]	许娟, 张党锋. 尼达尼布对肺纤维化小鼠肺功能及内质网应激反应的影响[J/OL]. 中华肺部疾病杂志(电子版), 2023, 16(05): 673-675.
[8]	王刚, 单伟根, 刘娟, 李长庆, 王新民. 常规治疗联合阿托伐他丁钙片对支气管扩张症临床分析[J/OL]. 中华肺部疾病杂志(电子版), 2023, 16(04): 538-540.
[9]	陈意志. 核磁共振钆造影剂导致的肾源性系统性纤维化[J/OL]. 中华肾病研究电子杂志, 2024, 13(06): 358-358.
[10]	杜霞, 马梦青, 曹长春. 造影剂诱导的急性肾损伤的发病机制及干预靶点研究进展[J/OL]. 中华肾病研究电子杂志, 2024, 13(05): 279-282.
[11]	桑田, 赵磊, 佟琰, 欧阳清, 陈香美. 急性肾损伤的内质网应激相关基因和通路的生物信息学分析[J/OL]. 中华肾病研究电子杂志, 2024, 13(01): 26-33.
[12]	林庆喜, 曾钦霖, 谢琛璠, 魏梁锋. 封闭式钻孔引流联合阿托伐他汀治疗慢性硬膜下血肿的疗效观察[J/OL]. 中华神经创伤外科电子杂志, 2024, 10(02): 91-96.
[13]	温绍敏, 王雅晳, 施依璐, 段莎莎, 云书荣, 张小杉. 靶向超声造影技术在动脉粥样硬化治疗中的应用进展[J/OL]. 中华临床医师杂志(电子版), 2024, 18(05): 496-499.
[14]	郭如烨, 孟黎明, 陈楠, 宋玉莹, 尹海燕, 郭岩. Apelin/APJ系统对帕金森病模型的神经保护作用及机制研究进展[J/OL]. 中华诊断学电子杂志, 2023, 11(04): 276-282.
[15]	于乐林, 尚海龙, 杜红娣, 王莺, 王一超, 徐长贺, 叶娟, 赵世伟, 郑芳慧, 沈慧, 沈海林. 基于CT平扫的影像组学特征预测急性大脑中动脉闭塞机械取栓术后造影剂外渗的价值[J/OL]. 中华脑血管病杂志(电子版), 2024, 18(03): 215-223.

阅读次数

全文

摘要

选择文件类型/文献管理软件名称

选择包含的内容

Atorvastatin decreased the contrast-induced renal tubular epithelial cell apoptosis through regulation of the PERK/eIF2α/CHOP signaling pathway

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

Atorvastatin decreased the contrast-induced renal tubular epithelial cell apoptosis through regulation of the PERK/eIF2α/CHOP signaling pathway