顺铂致新西兰兔急性肾损伤模型
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摘要
目的建立顺铂诱导急性肾损伤新西兰兔模型。方法 24只雄性新西兰兔按体质量随机分为对照组及顺铂低、中、高剂量组,分别按0. 0、1. 0、2. 0、4. 0 mg/kg体质量顺铂经耳缘静脉注射染毒,低剂量组连续染毒5 d,中剂量组连续染毒3 d,高剂量组单次染毒,均1次/d,对照组不予任何处理。在染毒前和染毒第1、3、5、7天经耳中动脉采血,并留取24 h尿液,以比色法检测血清肌酐(Scr)、尿素氮(BUN)水平,以电感耦合等离子体-质谱法测血浆铂、24 h尿铂和肾铂水平,以酶联免疫吸附实验法测24 h尿肾损伤分子-1(KIM-1)水平。实验结束后处死新西兰兔,取左肾进行病理组织学检查。结果染毒第7天时,低、中、高剂量组新西兰兔体质量均低于同时间点对照组和同组染毒前(P <0. 05)。3个剂量组新西兰兔在染毒第3天起Scr水平均高于同时间点对照组(P <0. 05);中、高剂量组新西兰兔在染毒第3、5天Scr水平均高于同时间点低剂量组(P <0. 05)。中、高剂量组新西兰兔在染毒第3和5天BUN水平均高于同时间点对照组和低剂量组(P <0. 05); 3个剂量组新西兰兔在染毒第7天BUN水平均高于同时间点对照组(P <0. 05)。3个剂量组新西兰兔在染毒第1、3、5、7天血浆铂及24 h尿铂水平均高于同时间点对照组(P <0. 05),且均高于同组染毒前水平(P <0. 05)。高剂量组新西兰兔在染毒第3天24 h尿KIM-1水平高于对照组(P <0. 05),中剂量组新西兰兔在染毒第5天24 h尿KIM-1水平高于对照组(P <0. 05)。3个剂量组新西兰兔肾铂水平均高于对照组(P <0. 05)。顺铂引起兔肾脏病理改变主要为肾小管扩张、蛋白管型、嗜碱性变和间质性肾炎。结论顺铂可诱导兔急性肾损伤,其损伤程度与染毒剂量相关。顺铂剂量为1. 0 mg/kg体质量连续染毒5 d的造模方式接近顺铂临床使用方式,推荐以此方式造模。
Objective To establish a New Zealand rabbit model of acute kidney injury induced by cisplatin. Methods A total of 24 male New Zealand rabbits were randomly divided into control group,low-,medium-and high-dose cisplatin group according to the body mass. Rabbits were injected with cisplatin at 0. 0,1. 0,2. 0,4. 0 mg/kg body weight by auricular vein. Rabbits in low-dose group was continuously injected for 5 days,medium-dose group was continuously injected for 3 days,and the high-dose group was injected for once per day. Rabbits in the control group did not receive any treatment. Blood was collected from the middle ear artery and 24 h urine was taken before exposure and on day 1,day 3,day 5 and day 7 of injection. The serum creatinine( Scr) and urea nitrogen( BUN) were detected by colorimetric method,and 24 h urine kidney injury molecule 1( KIM-1) was measured by enzyme-linked immunosorbent assay. Plasma platinum,24 h urinary platinum and renal platinum level were detected by inductively coupled plasma mass spectrometry.At the end of the experiment,rabbits were sacrificed and the left kidney was taken for histopathological examination.Results The body mass of rabbits of the low-,medium-and high-dose groups on day 7 after cisplatin exposure was lower than that of the control group( P < 0. 05),and lower than that of the same group before exposure( P < 0. 05). After 3 days of exposure,the Scr level in each dose group was higher than that of the control group( P < 0. 05),the Scr level on day 3and day 5 in medium-and high-dose groups were higher than that of the low-dose group( P < 0. 05). The BUN levels on day 3 and day 5 in medium-and high-dose group were higher than that of the control group and low-dose group( P <0. 05),the BUN levels on day 7 in three dose groups were higher than that of the control group( P < 0. 05). The levels of plasma platinum and 24 h urinary platinum in the three doses groups of New Zealand rabbits on day 1,day 3,day 5 and day 7 after exposure were higher than that of the control group( P < 0. 05),and were higher than the pre-treatment levels of the same group( P < 0. 05). The level of 24 h urinary KIM-1 in the meclium-dose group of New Zealand rabbits was higher than that of the control group on day 3 of exposure( P < 0. 05). The level of 24 h urinary KIM-1 in the mediumdose group of New Zealand rabbits on the 5th day after exposure was higher than that of the control group( P < 0. 05). The renal platinum levels in the three groups of New Zealand rabbits were higher than that in the control group( P < 0. 05).The pathological changes of rabbit kidney caused by cisplatin are mainly tubular dilatation,protein cast,alkalophilic and interstitial nephritis. Conclusion Cisplatin can induce acute kidney injury in rabbits,and the degree of injury is dosedependent. The dose of 1. 0 mg/kg body weight continuous injection for 5 days is closely related to clinical use of cisplatin,which is recommended for model establishment.
Objective To establish a New Zealand rabbit model of acute kidney injury induced by cisplatin. Methods A total of 24 male New Zealand rabbits were randomly divided into control group,low-,medium-and high-dose cisplatin group according to the body mass. Rabbits were injected with cisplatin at 0. 0,1. 0,2. 0,4. 0 mg/kg body weight by auricular vein. Rabbits in low-dose group was continuously injected for 5 days,medium-dose group was continuously injected for 3 days,and the high-dose group was injected for once per day. Rabbits in the control group did not receive any treatment. Blood was collected from the middle ear artery and 24 h urine was taken before exposure and on day 1,day 3,day 5 and day 7 of injection. The serum creatinine( Scr) and urea nitrogen( BUN) were detected by colorimetric method,and 24 h urine kidney injury molecule 1( KIM-1) was measured by enzyme-linked immunosorbent assay. Plasma platinum,24 h urinary platinum and renal platinum level were detected by inductively coupled plasma mass spectrometry.At the end of the experiment,rabbits were sacrificed and the left kidney was taken for histopathological examination.Results The body mass of rabbits of the low-,medium-and high-dose groups on day 7 after cisplatin exposure was lower than that of the control group( P < 0. 05),and lower than that of the same group before exposure( P < 0. 05). After 3 days of exposure,the Scr level in each dose group was higher than that of the control group( P < 0. 05),the Scr level on day 3and day 5 in medium-and high-dose groups were higher than that of the low-dose group( P < 0. 05). The BUN levels on day 3 and day 5 in medium-and high-dose group were higher than that of the control group and low-dose group( P <0. 05),the BUN levels on day 7 in three dose groups were higher than that of the control group( P < 0. 05). The levels of plasma platinum and 24 h urinary platinum in the three doses groups of New Zealand rabbits on day 1,day 3,day 5 and day 7 after exposure were higher than that of the control group( P < 0. 05),and were higher than the pre-treatment levels of the same group( P < 0. 05). The level of 24 h urinary KIM-1 in the meclium-dose group of New Zealand rabbits was higher than that of the control group on day 3 of exposure( P < 0. 05). The level of 24 h urinary KIM-1 in the mediumdose group of New Zealand rabbits on the 5th day after exposure was higher than that of the control group( P < 0. 05). The renal platinum levels in the three groups of New Zealand rabbits were higher than that in the control group( P < 0. 05).The pathological changes of rabbit kidney caused by cisplatin are mainly tubular dilatation,protein cast,alkalophilic and interstitial nephritis. Conclusion Cisplatin can induce acute kidney injury in rabbits,and the degree of injury is dosedependent. The dose of 1. 0 mg/kg body weight continuous injection for 5 days is closely related to clinical use of cisplatin,which is recommended for model establishment.
引文
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[9]公伟,刘丹,岳会敏,等.冬虫夏草菌丝体提取物抑制顺铂诱导的肾小管上皮细胞损伤[J].中国免疫学杂志,2016,32(5):669-672.
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[11]魏伟,吴希美,李元建.药理实验方法学[J].现代生物医学进展,2012,12(7):1380-1383.
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[13]丘岳,杨玉芳,黄振光,等.三七总皂苷对顺铂肾损害大鼠的保护作用[J].中国实验方剂学杂志,2016,22(21):104-109.
[14]杨慧海,王露露,何晓凤,等.梅花鹿鹿茸总蛋白对顺铂诱导肾损伤小鼠的保护作用及其机制[J].中国药理学与毒理学杂志,2016,31(6):561-566.
[15]原丽欣.五味子乙素对顺铂所致大鼠肾氧化损伤的保护作用[J].中国医院药学杂志,2014,34(10):826-828.
[16]曹思思,龚双,颜苗,等.肾复舒颗粒对顺铂肾毒性小鼠的保护作用研究[J].中国临床药理学杂志,2017,33(6):535-538.
[17]赵华叶,戚姝娅,李朝峰,等.艾迪注射液对顺铂所致大鼠肾毒性的保护作用研究[J].中国药学杂志,2016,51(22):1947-1951.
[18]舒婷婷.还原型谷胱甘肽对顺铂致大鼠急性肾损伤炎症反应的干预研究[D].南宁:广西医科大学,2014.
[19]KHWAJA A.Kdigo clinical practice guidelines for acute kidney injury[J].Nephron Clin Pract,2012,120(4):179-184.
[20]RYU H G,JEONG S J,KWON H Y,et al.Penta-O-galloyl-β-D-glucose attenuates cisplatin-induced nephrotoxicity via reactive oxygen species reduction in renal epithelial cells and enhances a.ntitumor activity in Caki-2 renal cance cells[J].Toxicol In Vitro,2012,26:206-214.
[21]WEI Q,DONG G,FRANKLIN J,et al.The pathological role of bax in cisplatin nephrotoxicity[J].Kidney Int,2007,72(1):53-62.
[22]ZHANG B,RAMESH G,UEMATSU S,et al.TLR4 signaling mediates inflammation and tissue injury in nephrotoxicity[J].J Am Soc Nephrol,2008,19(5):923-932.
[23]NEMATBAKHSH M,EBRAHIMIAN S,TOOYSERKANI M,et al.Gender difference in cisplatin-induced nephrotoxicity in arat model:greater intensity of damage in male than female[J].Nephro-Urology Monthly,2013,5(3):818-821.
[24]刘剑,徐韶东.常见铂类抗肿瘤药物不良反应的比较与防治[J].中国药房,2013,24(30):2857-2858.
[25]TOWNSEND D M,DENG M,ZHANG L,et al.Metabolism of cisplatin to a nephrotoxin in proximal tubule cells[J].J Am Soc Nephrol,2003,14(1):1-10.
[26]朱妍蒨,于锋,葛卫红.基于肾功能状况调整顺铂剂量的研究进展[J].药学与临床研究,2011,19(6):528-531.
[27]申俊,刘妍,张金晓,等.尿中肾损伤分子1水平升高对大鼠早期肾损伤的预测作用[J].中国药理学与毒理志,2012,26(2):212-217.
[2]MOTAMEDI F,NEMATBAKHSH M,MONAJEMI R.et al.Effect of pomegranate flower extract on cisplatin-induced nephrotoxicity in rats[J].J Nephropathol,2014,3(4):133-138.
[3]HANIGAN M H,DEVARAJAN P C.Cisplatin nephrotoxicity:molecular mechanisms[J].Cancer Therapy,2003,l:47-61.
[4]MOUSTAFA F E,SOBH M A,ABOUELKHEIR M,et al.Study of the effectof route of administrationof mesenchymal stem cells on cisplatin-induced acute kidney injury in spraguedawley rats[J].Int JStem Cells,2016,9(1):79-89.
[5]BULACIO R P,TORRES A M.Organic anion transporter 5(Oat5)renal expression and urinaryexcretion in rats treated with cisplatin:a potential biomarker of cisplatin-induced nephrotoxicity[J].Arch Toxicol,2013,87:1953-1962.
[6]CHOI D E,JIN Y J,LIM B J,et al.Pretreatment with darbepoetin attenuates renal injury in a rat model of cisplatin-induced nephrotoxicity[J].Korean J Intern Med,2009,24(3):238-246.
[7]KATAGIRI D,HAMASAKI Y,DOI K,et al.et al.Protection of glucagon-like peptide-1 in cisplatin-induced renal injury elucidates[J].J Am Soc Nephrol,2013,24(12):2034-2043.
[8]KIM H J,RAVICHANDRAN K,OZKOK A,et al.The watersoluble triptolide derivative pg490-88 protects against cisplatininduced acute kidney injury[J].J Pharmacol Exp Ther,2014,349(3):518-525.
[9]公伟,刘丹,岳会敏,等.冬虫夏草菌丝体提取物抑制顺铂诱导的肾小管上皮细胞损伤[J].中国免疫学杂志,2016,32(5):669-672.
[10]中华人民共和国卫生部合理用药专家委员会.中国医师药师临床用药指南[M].2版.重庆:重庆出版社,2014:483.
[11]魏伟,吴希美,李元建.药理实验方法学[J].现代生物医学进展,2012,12(7):1380-1383.
[12]陈浩,宓穗卿,赵威.药物引起肾功能损伤的研究进展[J].现代生物医学进展,2012,12(7):1380-1383.
[13]丘岳,杨玉芳,黄振光,等.三七总皂苷对顺铂肾损害大鼠的保护作用[J].中国实验方剂学杂志,2016,22(21):104-109.
[14]杨慧海,王露露,何晓凤,等.梅花鹿鹿茸总蛋白对顺铂诱导肾损伤小鼠的保护作用及其机制[J].中国药理学与毒理学杂志,2016,31(6):561-566.
[15]原丽欣.五味子乙素对顺铂所致大鼠肾氧化损伤的保护作用[J].中国医院药学杂志,2014,34(10):826-828.
[16]曹思思,龚双,颜苗,等.肾复舒颗粒对顺铂肾毒性小鼠的保护作用研究[J].中国临床药理学杂志,2017,33(6):535-538.
[17]赵华叶,戚姝娅,李朝峰,等.艾迪注射液对顺铂所致大鼠肾毒性的保护作用研究[J].中国药学杂志,2016,51(22):1947-1951.
[18]舒婷婷.还原型谷胱甘肽对顺铂致大鼠急性肾损伤炎症反应的干预研究[D].南宁:广西医科大学,2014.
[19]KHWAJA A.Kdigo clinical practice guidelines for acute kidney injury[J].Nephron Clin Pract,2012,120(4):179-184.
[20]RYU H G,JEONG S J,KWON H Y,et al.Penta-O-galloyl-β-D-glucose attenuates cisplatin-induced nephrotoxicity via reactive oxygen species reduction in renal epithelial cells and enhances a.ntitumor activity in Caki-2 renal cance cells[J].Toxicol In Vitro,2012,26:206-214.
[21]WEI Q,DONG G,FRANKLIN J,et al.The pathological role of bax in cisplatin nephrotoxicity[J].Kidney Int,2007,72(1):53-62.
[22]ZHANG B,RAMESH G,UEMATSU S,et al.TLR4 signaling mediates inflammation and tissue injury in nephrotoxicity[J].J Am Soc Nephrol,2008,19(5):923-932.
[23]NEMATBAKHSH M,EBRAHIMIAN S,TOOYSERKANI M,et al.Gender difference in cisplatin-induced nephrotoxicity in arat model:greater intensity of damage in male than female[J].Nephro-Urology Monthly,2013,5(3):818-821.
[24]刘剑,徐韶东.常见铂类抗肿瘤药物不良反应的比较与防治[J].中国药房,2013,24(30):2857-2858.
[25]TOWNSEND D M,DENG M,ZHANG L,et al.Metabolism of cisplatin to a nephrotoxin in proximal tubule cells[J].J Am Soc Nephrol,2003,14(1):1-10.
[26]朱妍蒨,于锋,葛卫红.基于肾功能状况调整顺铂剂量的研究进展[J].药学与临床研究,2011,19(6):528-531.
[27]申俊,刘妍,张金晓,等.尿中肾损伤分子1水平升高对大鼠早期肾损伤的预测作用[J].中国药理学与毒理志,2012,26(2):212-217.