摘要
目的:改进测定清醒小鼠肾小球滤过率(GFR)的异硫氰酸荧光素(FITC)标记的菊粉清除率检测方法,并应用于评估db/db小鼠28周龄内的GFR变化。方法:将FITC-菊粉以3.7μl/g体重的剂量对清醒小鼠进行内眦静脉丛匀速弹丸式注射,在注射后1、2、3、4、5、7、10、15、35、55、75min共11个时间点采集血标本检测荧光强度并计算GFR。10只8周雄性db/db小鼠及5只同窝雄性对照db/m小鼠,每4周检测体重、血糖及GFR至第28周,在20周和28周龄时检测尿糖、尿微量白蛋白并观察肾脏病理。结果:用改良方法测得的荧光值随时间变化的曲线符合两室模型,3min后缺失任意时间点的GFR差异无统计学意义(P>0.05)。db/db小鼠的体重、血糖均显著升高,GFR水平在16周龄为851.1±235.2μl/min,20周龄为796.0±338.7μl/min,显著高于对照组的440.5±182.1μl/min和299.6±74μl/min,差异均有统计学意义(P<0.05)。db/db小鼠在20周龄和28周龄尿微量白蛋白升高,并且出现肾小球肥大、系膜基质增多等病理表现。结论:改良版小鼠FITC-菊粉清除率检测法具有良好的重复性和可操作性。db/db小鼠在16~20周龄达到肾小球高滤过期,20周龄以后出现糖尿病肾病早期表现。
Objective:To improve the method of detecting fluorescein isothiocyanate(FITC)-insulin clearance which reflects glomerular filtration rate(GFR)of conscious mice,and to observe the hyperfiltration phase,the proteinuria and renal pathology of db/db mice.Methods:After bolus injection with FITC-insulin in,tail vein blood was collected in 1,2,3,4,5,7,10,15,35,55 and 75 minutes.Plasma fluorescence was tested and GFR was calculated.10Ten 8-week old male db/db mice and 5 their nondiabetic db/m littermates were tested for their weight,blood glucose and GFR in every 4 weeks.Urine microprotein and renal pathology were evaluated at 20-week and 28-week.Results:The plasma fluorescence data were fit to a two-phase exponential decay curve with good repeatability and feasibility. The body weight and blood glucose level wereas significantly higher in db/db mice. The GFRs wereas 851.1±235.2μl/min and 796.0±338.7μl/min in db/db mice at 16weeks and 20 weeks old,which were significantly higher than those of the control(P<0.05).The ratio of urine microprotein increased,and glomerular hypertrophy and enlargement of mesangial matrix were showed appeared in db/db mice at 20-week and 28-week.Conclusion:The improved FITC-insulin clearance detection method showed acceptable repeatability and feasibility,and db/db mice reached hyperfiltration phase when they were 16 to 20 weeks old.Other presentations of early diabetic nephropathy including microproteinuria,glomerular hypertrophy and enlargement of mesangial matrix were also presentedshowed in db/db mice.
作者
刘琳
姜世敏
张浩军
李鸿
杨悦
卓莉
李文歌
LIU Lin;JIANG Shi-min;ZHANG Hao-jun(Department of Nephrology,China-Japan Friendship Hospital,Beijing 100029,China)
出处
《中日友好医院学报》
CAS
2022年第4期221-225,238,F0002,共7页
Journal of China-Japan Friendship Hospital
基金
国家自然科学基金(81600547,81870495)
北京市自然科学基金(7202179)。
