镉暴露致大鼠肾损伤的量效关系及其机制

doi:10.3969/j.issn.1004-616x.2016.06.007

癌变·畸变·突变 ›› 2016, Vol. 28 ›› Issue (6): 446-452.doi: 10.3969/j.issn.1004-616x.2016.06.007

镉暴露致大鼠肾损伤的量效关系及其机制

王乐乐, 刘江正, 刘萌萌, 孔德钦, 张涛, 于卫华, 刘瑞, 张晓迪, 王欣, 海春旭

第四军医大学预防医学院毒理学教研室, 陕西省自由基生物学与医学重点实验室, 陕西西安 710032

收稿日期:2016-07-19 修回日期:2016-09-28 出版日期:2016-11-30 发布日期:2016-11-30
通讯作者: 海春旭,E-mail:cx-hai@fmmu.edu.cn E-mail:cx-hai@fmmu.edu.cn
作者简介:王乐乐,E-mail:diana1147824730@163.com。
基金资助:
国家自然科学基金（81473010）；陕西省自然科学基金（2016JM8024）

Dose-dependent induction of renal injury by cadmium in rats

WANG Lele, LIU Jiangzheng, LIU Mengmeng, KONG Deqin, ZHANG Tao, YU Weihua, LIU Rui, ZHANG Xiaodi, WANG Xin, HAI Chunxu

Department of Toxicology, School of Public Health, Fourth Military Medical University, Shaanxi Key Lab of Free Radical Biology and Medicine, Xi'an 710032, Shaanxi, China

Received:2016-07-19 Revised:2016-09-28 Online:2016-11-30 Published:2016-11-30

摘要/Abstract

摘要： 目的：探讨不同剂量氯化镉（CdCl₂）染毒对大鼠肾损伤的量效关系及其机制。方法：雄性SD大鼠40只，随机分为0、1、3和5mg/kg CdCl₂染毒组，每天灌胃1次。连续染毒4周后检测血清尿素氮（BUN）和肌酐（SCr）水平评价肾功能；HE染色观察肾脏病理损伤；透射电镜观察肾组织超微结构变化；DHE及MitoSOX荧光探针检测肾组织中活性氧（ROS）含量变化；免疫印迹检测SOD1、SOD2、GPx-1、CAT、Bax、Bcl-2以及GRP78蛋白的表达。结果：与对照组比较，镉暴露后大鼠体质量及肾体比差异无统计学意义（P>0.05）；随CdCl₂剂量增加，血清BUN含量逐渐增加（r=0.463，P<0.05）；肾脏病理损伤进行性加重，出现肾小球肾小管肿胀，肾小管管腔狭窄、上皮细胞坏死脱落堆积于管腔、管腔内可见管型，肾间质充血，炎细胞浸润；超微结构观察显示，肾小管上皮细胞线粒体损伤，肿胀、变形、空泡样变程度逐渐加重；肾组织中ROS含量逐渐增加；抗氧化酶SOD2、GPx-1和CAT表达逐渐增加（r依次为0.854、0.975、0.918，P均<0.05），SOD1表达逐渐减少（r=-0.987，P<0.05）；肾脏组织促凋亡蛋白Bax表达在0~3 mg/kg CdCl₂处理组逐渐增加（r=0.226，P<0.05），抑凋亡蛋白Bcl-2表达逐渐减少（r=-0.85，P<0.05），Bax/Bcl-2呈剂量依赖性升高（r=0.83，P<0.05）；同时，内质网应激标志蛋白GRP78的表达也随CdCl₂剂量增加而升高（r=0.913，P<0.05）。结论：不同剂量镉暴露致雄性SD大鼠肾脏损伤存在剂量效应关系，其作用机制可能是镉暴露导致氧化还原稳态失衡，从而引起氧化应激，进一步引起凋亡和组织损伤。

关键词: 镉暴露, 肾损伤, 剂量效应关系, 氧化应激, 活性氧

Abstract: OBJECTIVE: This study was designed to investigate the dose-dependent relationship and the possible mechanisms of renal injury by cadmium in rats. METHODS: Forty male rats were randomly divided into four groups (0,1,3 and 5 mg/kg CdCl₂) groups. The rats were intra-gastrically administrated CdCl₂ daily to investigate the induction of renal injury. Blood urea nitrogen (BUN) and serum creatinine (SCr) were measured to evaluate renal function. Kidney tissues were stained with hematoxylin-eosin (HE) and observed for histopathological changes. Transmission electron microscopye was used to detect ultrastructural abnormalities. Levels of ROS in kidney was detected by DHE and MitoSOX staining. The levels of SOD1,SOD2,GPx-1,CAT,Bax,Bcl-2,GRP78 were measured by Western blotting. RESULTS: After exposure to CdCl₂ for 4 weeks,the relative kidney weights showed no significant difference among the 4 groups. As the dose of CdCl₂ increased,the levels of BUN elevated gradually (r=0.463,P<0.05). Pathological damages to kidneys were progressively aggravated. Swelling of glomerulus and renal tubule,and necrosis of tubular epithelial cells were observed under light microscopy. And it was also observed that the lumina of renal tubules contained necrotic epithelial cellular debris and casts. Interstitial hyperemia and inflammatory infiltration were found as well. The ultrastructure of mitochondria in the epithelium of proximal convoluted tubule showed swelling,deformation and vacuolation in a dose-dependent manner. The level of kidney ROS elevated gradually. Levels of SOD2,GPx-1 and CAT were upregulated (r=0.854,0.975,0.918,P all<0.05) while SOD1 were downregulated (r=-0.987,P<0.05) as the dose of CdCl₂ increased. Meanwhile,levels of the apoptosis-promoting Bax increased (r=0.226,P<0.05) while the anti- apoptotic Bcl-2 decreased (r=-0.85,P<0.05) and the ratio of Bax/Bcl-2 increased (r=0.83,P<0.05). The levels of GRP78 also increased when the dose of CdCl₂ increased (r=0.913,P<0.05). CONCLUSION: The results demonstrate that there was a dose-dependent toxic effect of CdCl₂ on renal structure and function. Moreover,oxidative stress might be the key mechanism involved in the toxicity of cadmium because of increased antioxidase expression and levels of ROS.

Key words: cadmium exposure, renal injury, dose-dependent, oxidative stress, reactive oxygen species

中图分类号:

R994.6

王乐乐, 刘江正, 刘萌萌, 孔德钦, 张涛, 于卫华, 刘瑞, 张晓迪, 王欣, 海春旭. 镉暴露致大鼠肾损伤的量效关系及其机制[J]. 癌变·畸变·突变, 2016, 28(6): 446-452.

WANG Lele, LIU Jiangzheng, LIU Mengmeng, KONG Deqin, ZHANG Tao, YU Weihua, LIU Rui, ZHANG Xiaodi, WANG Xin, HAI Chunxu. Dose-dependent induction of renal injury by cadmium in rats[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2016, 28(6): 446-452.

参考文献

[1] Johri N,Jacquillet G,Unwin R. Heavy metal poisoning:the effects of cadmium on the kidney[J]. Biometals,2010, 23(5):783-792.
[2] Yang H,Shu Y. Cadmium transporters in the kidney and cadmium-induced nephrotoxicity[J]. Int J Mol Sci,2015, 16(1):1484-1494.
[3] Wang Y,Wu Y,Luo K,et al. The protective effects of selenium on cadmium-induced oxidative stress and apoptosis via mitochondria pathway in mice kidney[J]. Food Chem Toxicol, 2013,58:61-67.
[4] Wang C,Blough ER,Arvapalli R,et al. Metabolic syndrome-induced tubulointerstitial injury:Role of oxidative stress and preventive effects of acetaminophen[J]. Free Radic Biol Med,2013,65:1417-1426.
[5] Mitchell T,Saba H,Laakman J,et al. Role of mitochondrial-derived oxidants in renal tubular cell cold-storage injury[J]. Free Radic Biol Med,2010,49(8):1273-1282.
[6] Latchoumycandane C,Nagy LE,McIntyre TM. Chronic ethanol ingestion induces oxidative kidney injury through taurine-inhibitable inflammation[J]. Free Radic Biol Med, 2014,69:403-416.
[7] Xu W,Neckers L. Gr(i)p the ER to stress out melanoma[J]. Cancer Cell,2016,29(6):769-771.
[8] Chen S,Ren Q,Zhang J,et al. N-acetyl-L-cysteine protects against cadmium-induced neuronal apoptosis by inhibiting ROS-dependent activation of Akt/mTOR pathway in mouse brain[J]. Neuropath Appl Neuro,2014,40(6):759-777.
[9] Zhai Q,Narbad A,Chen W. Dietary strategies for the treatment of cadmium and lead toxicity[J]. Nutrients,2015, 7(1):552-571.
[10] Bernhoft RA. Cadmium toxicity and treatment[J]. Sci World J,2013,2013:1-7.
[11] Baba H, Tsuneyama K, Kumada T, et al. Histopathological analysis for osteomalacia and tubulopathy in itai-itai disease[J]. J Toxicol Sci,2014,39(1):91-96.
[12] Adiele RC,Stevens D,Kamunde C. Features of cadmium and calcium uptake and toxicity in rainbow trout (Oncorhynchus mykiss) mitochondria[J]. Toxicol in Vitro,2012,26(1):164-173.
[13] Jorge-Nebert LF,Galvez-Peralta M,Landero Figueroa J,et al. Comparing gene expression during cadmium uptake and distribution:untreated versus oral Cd-treated wild-type and ZIP14 knockout mice[J]. Toxicol Sci,2014,143(1):26-35.
[14] Prozialeck WC,Edwards JR. Mechanisms of cadmium-induced proximal tubule injury:new insights with implications for biomonitoring and therapeutic interventions[J]. J Pharmacol Exp Ther,2012,343(1):2-12.
[15] Matović V,Buha A,Bulat Z,et al. Cadmium toxicity revisited:focus on oxidative stress induction and interactions with zinc and magnesium[J]. Arch Toxicol,2011,62(1):65-75.
[16] Wang J,Zhu H,Liu X,et al. N-acetylcysteine protects against cadmium-induced oxidative stress in rat hepatocytes[J]. J Vet Sci,2014,15(4):485.
[17] 海春旭. 自由基医学[M]. 西安: 第四军医大学出版社, 2006.
[18] Ben P,Zhang Z,Xuan C,et al. Protective effect of l-theanine on cadmium-induced apoptosis in PC12 cells by inhibiting the mitochondria-mediated pathway[J]. Neurochem Res,2015, 40(8):1661-1670.
[19] Luevano J,Damodaran C. A review of molecular events of cadmium-induced carcinogenesis[J]. J Environ Pathol Toxicol Oncol,2014,33(3):183-194.

镉暴露致大鼠肾损伤的量效关系及其机制

Dose-dependent induction of renal injury by cadmium in rats

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	艾多, 徐安琦, 孔德钦, 刘颖, 于卫华, 王钊, 彭洁, 刘瑞, 张晓迪, 海春旭, 李文丽, 王欣, 刘江正. 催化性抗氧化剂AEOL-10150对氮芥诱导小鼠急性肝损伤的保护作用[J]. 癌变·畸变·突变, 2022, 34(1): 40-46,61.
[2]	尹亚萍, 王宇, 田雨阳, 马焜, 李百祥. 褪黑素在帕金森病治疗中的研究进展[J]. 癌变·畸变·突变, 2022, 34(1): 75-78,81.
[3]	曹家豪, 彭越, 段佳琪, 刘桥松, 刘启玲, 秦绪军. 酒精性脂肪肝的线粒体作用机制研究进展[J]. 癌变·畸变·突变, 2021, 33(6): 475-481.
[4]	李宏伟, 孔德钦, 于卫华, 吴昊, 王钊, 刘瑞, 海春旭, 王欣, 刘江正, 李文丽. 齐墩果酸对酒精诱导的大鼠胃壁氧化损伤的保护作用[J]. 癌变·畸变·突变, 2021, 33(5): 365-369.
[5]	于卫华, 孔德钦, 龙子, 刘瑞, 李文丽, 海春旭. 抗氧化悖论真的成立吗？[J]. 癌变·畸变·突变, 2021, 33(5): 388-392.
[6]	蒋杉, 杜凤, 康秉文, 殷草草, 周健, 石莹, 王玥, 周耿瑶, 秦绪军. 异鼠李素通过减轻氧化应激延缓D-半乳糖诱导的人脐静脉内皮细胞衰老[J]. 癌变·畸变·突变, 2021, 33(2): 95-100.
[7]	陈廷玉, 陈大印, 沈洪, 富徐燕, 卢春凤. 槲皮素对异烟肼诱导肝细胞毒性的保护作用及其机制[J]. 癌变·畸变·突变, 2021, 33(1): 12-16.
[8]	赵俊伟, 华辰凤, 尚平平, 谢复炜, 李翔. 亚砷酸As (Ⅲ)诱导人肺癌A549细胞氧化应激反应的实验研究[J]. 癌变·畸变·突变, 2021, 33(1): 28-31.
[9]	张力, 张爱华, 王文娟. 甲基汞暴露致学习记忆功能障碍作用机制的研究进展[J]. 癌变·畸变·突变, 2020, 32(6): 490-493.
[10]	孙娇娇, 车碧众, 罗秋林, 翟兵中, 信丽丽. 1,25-二羟基维生素D₃对PM_2.5所致HBE细胞氧化损伤的保护作用[J]. 癌变·畸变·突变, 2020, 32(2): 92-97.
[11]	师盼, 林重华, 舒易方, 瞿家权, 宋淑亮. 柠檬酸转运体SLC25A1对食管鳞癌细胞体外放射敏感性的影响及其分子机制[J]. 癌变·畸变·突变, 2020, 32(2): 132-138.
[12]	金磊, 周珈右, 韩家诚, 管浩军, 王帅, 彭洁, 王欣, 海春旭, 李文丽. 降脂酮对棕榈酸诱导BRL-3A细胞胰岛素抵抗的保护作用[J]. 癌变·畸变·突变, 2020, 32(1): 21-28.
[13]	张旭光, 杜瑞琥, 陈彦平, 赵艳. 维生素E琥珀酸酯通过酸性神经磷脂酶-神经酰胺诱导胃癌细胞凋亡的机制研究[J]. 癌变·畸变·突变, 2019, 31(5): 379-384.
[14]	徐海明, 金林, 张凡, 史新琛, 邢娜, 汪岭, 张鹏举, 德小明, 张银凤. 改进型生物慢滤池对饮用水中氟及相关污染物的去除效果研究[J]. 癌变·畸变·突变, 2019, 31(3): 238-241,248.
[15]	廖鼐, 龙子, 海春旭, 王欣. 2,3,7,8-四氯二苯并二噁英染毒或联合高脂饮食致小鼠糖脂代谢紊乱的实验研究[J]. 癌变·畸变·突变, 2019, 31(2): 111-118.