尿微量蛋白在对比剂肾病早期肾损伤中的变化及意义
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摘要
目的探讨尿液中β-2微球蛋白(β2-MG)、Tamm-Horsfall蛋白(THP)、肾损伤分子-1(KIM-1)、血肌酐(Scr)等指标,在对比剂肾病(CIN)模型大鼠早期肾损伤中的变化及意义。方法选择成年雄性大鼠50只,随机分为实验组和对照组,受试大鼠禁食水12 h后,经尾静脉注射吲哚美辛10 mg/kg,15 min后注射左旋硝基精氨酸甲酯10 mg/kg;再过15 min注射碘必乐(3 g I/kg),采用该方法建立造影剂肾病大鼠模型30例(实验组),对照组20例将碘必乐改为相同体积0. 9%氯化钠溶液,其余处理相同。分别于造模前、造模后2 h、6 h、12 h、24 h、48 h经大鼠内眦静脉抽血1 ml,4℃以3 000 r/min离心10 min,取血清测定SCr,代谢笼收集尿液,以3 000 r/min离心20 min后取上清,测定各时间点尿β2-MG、尿THP、尿KIM-1。在各个时间点将实验组大鼠各取2只,脱臼处死后取肾脏,制作肾脏组织切片,400倍光学显微镜下观察大鼠肾脏病理变化。结果大鼠在造模后12 h、24 h、48 h时,实验组的尿β2-MG浓度高于对照组,差异有统计学意义(P <0. 05);造模后24 h、48 h时,实验组的尿THP浓度高于对照组,差异有统计学意义(P <0. 05);造模后6 h、12 h、24 h、48 h时,实验组的尿KIM-1浓度高于对照组,差异有统计学意义(P <0. 05);造模后24 h、48 h时,实验组的Scr浓度高于对照组,差异有统计学意义(P <0. 05)。此外,大鼠肾脏在造模后2 h开始出现病理变化,并在12 h左右最为明显,在造模后12 h时,大鼠肾脏中脱落的肾小管细胞数量增加,小管结构紊乱、破坏。结论反映近端肾小管损伤的指标尿KIM-1、尿β2-MG,在对比剂肾病早期肾损伤的识别中具有较高价值。
Objective To investigate the changes and significance of β2-microglobulin( β2-MG),tamm-horsfall protein( THP),kidney injury molecule-1( KIM-1) and Serum creatinine( Scr) in early renal injury of contrast-induced nephropathy( CIN) in rats. Methods A total of 50 adult male SD rats were randomly divided into experimental group( n =30) and control group( n = 20). After 12-hour fasting,the rats were injected with indomethacin 10mg/kg via tail vein,15 min later,were injected with L-NAME 10mg/kg,15 min later,were injected with iodobiline 3mg/kg. The 30 rat models of contrast medium nephropathy were established by this method( experimental group). The rats in control group were given the same volume of normal saline in stead of iodobilol. Before establishing the models and at 2h,6h,12 h,24h,48 h after establishing models,the vein blood specimens were collected to detect the levels of SCr,and the urine specimens were collected to detect the levels of β2-MG,THP,KIM-1 in different time points. The two rats in experimental group in different time points were sacrificed,and the kidneys were taken out to make the kidneys tissue section,then the pathological changes of kidney were observed under the optical microscope( 400 ×). Results Th elevels β2-MG in urine of rats at 12 h,24h,48 h after establishing the models were significantly higher than those in control group( P < 0. 05). At 24 h,48h after establishing the models,the urine levels of THP in experimental group were significantly higher than those in control group( P < 0. 05). The urine levels of KIM-1 at 6h,12 h,24h,48 h after establishing the models in experimental group were significantly higher than those in control group( P < 0. 05). In additionthe levels of Scr at 24 h,48h after establishing the models in experimental group were significantly higher than those in control group( P < 0. 05). Moreover the pathological changes of kidney were observed at2 h after establishing the models in experimental group,which were most obvious at 12 h,at the same time,the tubular cells shed in the kidney were increasing,the organization of tubules was disorganized and destroyed. Conclusion The urine KIM-1and β2-MG can reflex the impairment of proximal tubule in rats with contrast-agent nephropathy,which is of great value in early diagnosis of renal injury of contrast-agent nephropathy.
Objective To investigate the changes and significance of β2-microglobulin( β2-MG),tamm-horsfall protein( THP),kidney injury molecule-1( KIM-1) and Serum creatinine( Scr) in early renal injury of contrast-induced nephropathy( CIN) in rats. Methods A total of 50 adult male SD rats were randomly divided into experimental group( n =30) and control group( n = 20). After 12-hour fasting,the rats were injected with indomethacin 10mg/kg via tail vein,15 min later,were injected with L-NAME 10mg/kg,15 min later,were injected with iodobiline 3mg/kg. The 30 rat models of contrast medium nephropathy were established by this method( experimental group). The rats in control group were given the same volume of normal saline in stead of iodobilol. Before establishing the models and at 2h,6h,12 h,24h,48 h after establishing models,the vein blood specimens were collected to detect the levels of SCr,and the urine specimens were collected to detect the levels of β2-MG,THP,KIM-1 in different time points. The two rats in experimental group in different time points were sacrificed,and the kidneys were taken out to make the kidneys tissue section,then the pathological changes of kidney were observed under the optical microscope( 400 ×). Results Th elevels β2-MG in urine of rats at 12 h,24h,48 h after establishing the models were significantly higher than those in control group( P < 0. 05). At 24 h,48h after establishing the models,the urine levels of THP in experimental group were significantly higher than those in control group( P < 0. 05). The urine levels of KIM-1 at 6h,12 h,24h,48 h after establishing the models in experimental group were significantly higher than those in control group( P < 0. 05). In additionthe levels of Scr at 24 h,48h after establishing the models in experimental group were significantly higher than those in control group( P < 0. 05). Moreover the pathological changes of kidney were observed at2 h after establishing the models in experimental group,which were most obvious at 12 h,at the same time,the tubular cells shed in the kidney were increasing,the organization of tubules was disorganized and destroyed. Conclusion The urine KIM-1and β2-MG can reflex the impairment of proximal tubule in rats with contrast-agent nephropathy,which is of great value in early diagnosis of renal injury of contrast-agent nephropathy.
引文
1 Stacul F,van der Molen AJ,Reimer P,et al. Contrast induced nephropathy:updated ESUR contrast media safety committee guidelines.Eur Radiol,2011,21:2527-2541.
2 吴凤丽,马晓光.联合检测血清CysC、血清hs CRP、和尿β2-MG在2型糖尿病早期肾病中的诊断价值.中国实验诊断学,2014,35:778-780.
3 Lane BR. Molecular markers of kidney injury. Urol Oncol,2013,31:682-5.
4 Gutierrez-Escolano A,Santacruz-Vazquez E,Gomez-Perez F. Dysregulated microRNAs involved in contrast-induced acute kidney injury in rat and human. Ren Fail,2015,37:1498-1506.
5 Ozcan OU,Adanir Er H,Gulec S,et al. Impact of Metabolic Syndrome on Development of Contrast-Induced Nephropathy After Elective Percutaneous Coronary Intervention Among Nondiabetic Patients. Clin Cardiol,2015,38:150-156.
6 Limbruno U,Picchi A,Micheli A,et al. Refining the assessment of contrast-induced acute kidney injury:the load-to-damage relationship. J Cardiovasc Med(Hagerstown),2013,15:587.
7 Won AJ,Kim S,Kim YG,et al. Discovery of urinary metabolomic biomarkers for early detection of acute kidney injury. Mol Biosyst,2016,12:133-144.
8 Schrezenmeier EV,Barasch J,Budde K,et al. Biomarkers in acute kidney injury-pathophysiological basis and clinical performance. Actaphysiol,2017,219:556-574.
9 Duncan L,Heathcote J,Djurdjev O,et al. Screening for renal disease using serum creatinine:who are we missing. Nephrol Dial Transplant,2001,16:1042-1046.
10 Swedko PJ,Clark HD,Paramsothy K,et al. Serum creatinine is an inadequate screening test for renal failure in elderly patients. Arch Intern Med,2003,163:356-360.
11 Vercellino M,Bezante GP,Balbi M. Contrast medium induced nephropathy:new insights into prevention and risk management. Cardiovasc Hematol Agents Med Chem,2009,7:166-180.
12 李东韬,郑建勇,赵力,等.β2微球蛋白、KIM-1和NGAL对大鼠对比剂肾病模型早期诊断的价值.医药论坛杂志,2015,36:17-19.
13 魏宇鹏,张季,杨金萍.血胱抑素C和尿中4种蛋白联合检测对早期糖尿病肾病的诊断价值.中国临床研究,2011,24:722-722.
14 刘文花,刘海霞,胡文博.糖尿病肾病患者血清胱抑素C、β2-微球蛋白水平及其与e GFR的相关性.中国老年学,2015,35:3002-3003.
15 付锴,赵久阳,滕思远.急性肾损伤早期生物标志物研究进展.大连医科大学学报,2016,38:392-396.
16 van Timmeren MM,van den Heuvel MC,Bailly V,et al. Tubular kidney injury molecule-1(KIM-1)in human renal disease. J Pathol,2007,212:209-217.
17 Ma X,Zhang BR,Li DT. Value of urinary kidney injury molecule-1protein in early diagnosis of radiocontrast-induced nephropathy in rats.Nan Fang Yi Ke Da Xue Xue Bao,2011,31:357-360.
18 李慧凛,张金元.肾损伤分子1在低渗造影剂肾病模型大鼠肾组织的表达及缺氧诱导因子1α对其的影响.肾脏病与透析肾移植杂志,2010,19:42-49.
19 Vlasakova K,Erdos Z,Troth SP,et al. Evaluation of the relative performance of 12 urinary biomarkers for renal safety across 22 rat sensitivity and specificity studies. Toxicol Sci,2014,138:3.
2 吴凤丽,马晓光.联合检测血清CysC、血清hs CRP、和尿β2-MG在2型糖尿病早期肾病中的诊断价值.中国实验诊断学,2014,35:778-780.
3 Lane BR. Molecular markers of kidney injury. Urol Oncol,2013,31:682-5.
4 Gutierrez-Escolano A,Santacruz-Vazquez E,Gomez-Perez F. Dysregulated microRNAs involved in contrast-induced acute kidney injury in rat and human. Ren Fail,2015,37:1498-1506.
5 Ozcan OU,Adanir Er H,Gulec S,et al. Impact of Metabolic Syndrome on Development of Contrast-Induced Nephropathy After Elective Percutaneous Coronary Intervention Among Nondiabetic Patients. Clin Cardiol,2015,38:150-156.
6 Limbruno U,Picchi A,Micheli A,et al. Refining the assessment of contrast-induced acute kidney injury:the load-to-damage relationship. J Cardiovasc Med(Hagerstown),2013,15:587.
7 Won AJ,Kim S,Kim YG,et al. Discovery of urinary metabolomic biomarkers for early detection of acute kidney injury. Mol Biosyst,2016,12:133-144.
8 Schrezenmeier EV,Barasch J,Budde K,et al. Biomarkers in acute kidney injury-pathophysiological basis and clinical performance. Actaphysiol,2017,219:556-574.
9 Duncan L,Heathcote J,Djurdjev O,et al. Screening for renal disease using serum creatinine:who are we missing. Nephrol Dial Transplant,2001,16:1042-1046.
10 Swedko PJ,Clark HD,Paramsothy K,et al. Serum creatinine is an inadequate screening test for renal failure in elderly patients. Arch Intern Med,2003,163:356-360.
11 Vercellino M,Bezante GP,Balbi M. Contrast medium induced nephropathy:new insights into prevention and risk management. Cardiovasc Hematol Agents Med Chem,2009,7:166-180.
12 李东韬,郑建勇,赵力,等.β2微球蛋白、KIM-1和NGAL对大鼠对比剂肾病模型早期诊断的价值.医药论坛杂志,2015,36:17-19.
13 魏宇鹏,张季,杨金萍.血胱抑素C和尿中4种蛋白联合检测对早期糖尿病肾病的诊断价值.中国临床研究,2011,24:722-722.
14 刘文花,刘海霞,胡文博.糖尿病肾病患者血清胱抑素C、β2-微球蛋白水平及其与e GFR的相关性.中国老年学,2015,35:3002-3003.
15 付锴,赵久阳,滕思远.急性肾损伤早期生物标志物研究进展.大连医科大学学报,2016,38:392-396.
16 van Timmeren MM,van den Heuvel MC,Bailly V,et al. Tubular kidney injury molecule-1(KIM-1)in human renal disease. J Pathol,2007,212:209-217.
17 Ma X,Zhang BR,Li DT. Value of urinary kidney injury molecule-1protein in early diagnosis of radiocontrast-induced nephropathy in rats.Nan Fang Yi Ke Da Xue Xue Bao,2011,31:357-360.
18 李慧凛,张金元.肾损伤分子1在低渗造影剂肾病模型大鼠肾组织的表达及缺氧诱导因子1α对其的影响.肾脏病与透析肾移植杂志,2010,19:42-49.
19 Vlasakova K,Erdos Z,Troth SP,et al. Evaluation of the relative performance of 12 urinary biomarkers for renal safety across 22 rat sensitivity and specificity studies. Toxicol Sci,2014,138:3.