CAJ | 학술논문

目的建立三聚氰胺泌尿系结石大鼠模型，观察肾脏病理改变，初步探讨三聚氰胺亚慢性肾脏毒性的可能机制。方法将3周龄刚断乳雄性Wistar大鼠60只按随机数字表法分为实验组2组（A、B组）和对照组（C组），每组20只，A、B、C组分别给予三聚氰胺质量分数为1％、2％、0的饲料。比较大鼠体重、摄食量、血清尿素氮和肌酐水平。15周时（用药期末）3组各随机选取半数处死，剩余半数停药继续喂养4周后（停药期末）处死，均行肾脏重量、系数比较并观察病理改变。采用高效液相色谱-质谱联用法检测血清和结石中三聚氰胺、三聚氰酸和尿酸含量。结果用药期末，A、B组血清尿素氮水平分别为( 13.23 ±5.10)、( 18.30 ±5.90) mmol/L,均高于C组(8.23 ±2.30) mmol/L(P＜0.01)；B组肌酐水平(19.90 ±2.90) mmoL/L高于C组(10.04±1.73) mmol/L(P＜0.01)；A、B组左右肾系数均高于C组(P＜0.01)；肾脏中可见结晶形成，结晶所在肾小管扩张明显，肾小管周围肾间质淋巴细胞浸润及间质纤维化，B组最为显著。停药期末，A、B组血清尿素氮水平分别为(17.96±2.04)、(19.20±3.36) mmol/L，均高于C组的（8.30±1.79）mmol/L(均P＜0.01)；B组肌酐水平(24.20±5.28) mmol/L高于C组(9.87 ±2.71) mmol/L(P ＜0.01)，且分别较用药期水平升高（分别为P＜0.01,P＜0.05）；两实验组肾脏系数较前下降；肾脏仍有结晶未排出，间质改变较停药前未见减轻。结论(1)饲喂3周龄刚断乳雄性Wistar大鼠含三聚氰胺质量分数为2％的饲料15周能够建立三聚氰胺泌尿系结石模型。饲喂含三聚氰胺质量分数为1％的饲料15周，仅少数大鼠可形成肉眼可见的肾脏结石，部分形成镜下结晶。结石成分主要为三聚氰胺(＞90％)、少量尿酸及极少量三聚氰酸。(2)三聚氰胺可导致大鼠肾功能受损，镜下可见肾小管结晶形成、肾小管扩张、炎症、纤维化等病理表现，并具有剂量依赖性。三聚氰胺的肾脏损害与结晶、结石梗阻有一定相关性。(3)停止摄入三聚氰胺后，肾功能及肾脏炎症、纤维化等改变短期内未见好转，结晶自行排出的过程较为缓慢，如结晶持续存在可能会造成进一步的或不可逆转的肾脏损伤。目的建立三聚氰胺泌尿繫結石大鼠模型，觀察腎髒病理改變，初步探討三聚氰胺亞慢性腎髒毒性的可能機製。方法將3週齡剛斷乳雄性Wistar大鼠60隻按隨機數字錶法分為實驗組2組（A、B組）和對照組（C組），每組20隻，A、B、C組分彆給予三聚氰胺質量分數為1％、2％、0的飼料。比較大鼠體重、攝食量、血清尿素氮和肌酐水平。15週時（用藥期末）3組各隨機選取半數處死，剩餘半數停藥繼續餵養4週後（停藥期末）處死，均行腎髒重量、繫數比較併觀察病理改變。採用高效液相色譜-質譜聯用法檢測血清和結石中三聚氰胺、三聚氰痠和尿痠含量。結果用藥期末，A、B組血清尿素氮水平分彆為( 13.23 ±5.10)、( 18.30 ±5.90) mmol/L,均高于C組(8.23 ±2.30) mmol/L(P＜0.01)；B組肌酐水平(19.90 ±2.90) mmoL/L高于C組(10.04±1.73) mmol/L(P＜0.01)；A、B組左右腎繫數均高于C組(P＜0.01)；腎髒中可見結晶形成，結晶所在腎小管擴張明顯，腎小管週圍腎間質淋巴細胞浸潤及間質纖維化，B組最為顯著。停藥期末，A、B組血清尿素氮水平分彆為(17.96±2.04)、(19.20±3.36) mmol/L，均高于C組的（8.30±1.79）mmol/L(均P＜0.01)；B組肌酐水平(24.20±5.28) mmol/L高于C組(9.87 ±2.71) mmol/L(P ＜0.01)，且分彆較用藥期水平升高（分彆為P＜0.01,P＜0.05）；兩實驗組腎髒繫數較前下降；腎髒仍有結晶未排齣，間質改變較停藥前未見減輕。結論(1)飼餵3週齡剛斷乳雄性Wistar大鼠含三聚氰胺質量分數為2％的飼料15週能夠建立三聚氰胺泌尿繫結石模型。飼餵含三聚氰胺質量分數為1％的飼料15週，僅少數大鼠可形成肉眼可見的腎髒結石，部分形成鏡下結晶。結石成分主要為三聚氰胺(＞90％)、少量尿痠及極少量三聚氰痠。(2)三聚氰胺可導緻大鼠腎功能受損，鏡下可見腎小管結晶形成、腎小管擴張、炎癥、纖維化等病理錶現，併具有劑量依賴性。三聚氰胺的腎髒損害與結晶、結石梗阻有一定相關性。(3)停止攝入三聚氰胺後，腎功能及腎髒炎癥、纖維化等改變短期內未見好轉，結晶自行排齣的過程較為緩慢，如結晶持續存在可能會造成進一步的或不可逆轉的腎髒損傷。
목적 건립삼취청알비뇨계결석대서모형，관찰신장병리개변，초보탐토삼취청알아만성신장독성적가능궤제。방법 장3주령강단유웅성Wistar대서60지안수궤수자표법분위실험조2조（A、B조）화대조조（C조），매조20지，A、B、C조분별급여삼취청알질량분수위1％、2％、0적사료。비교대서체중、섭식량、혈청뇨소담화기항수평。15주시（용약기말）3조각수궤선취반수처사，잉여반수정약계속위양4주후（정약기말）처사，균행신장중량、계수비교병관찰병리개변。채용고효액상색보-질보련용법검측혈청화결석중삼취청알、삼취청산화뇨산함량。결과 용약기말，A、B조혈청뇨소담수평분별위( 13.23 ±5.10)、( 18.30 ±5.90) mmol/L,균고우C조(8.23 ±2.30) mmol/L(P＜0.01)；B조기항수평(19.90 ±2.90) mmoL/L고우C조(10.04±1.73) mmol/L(P＜0.01)；A、B조좌우신계수균고우C조(P＜0.01)；신장중가견결정형성，결정소재신소관확장명현，신소관주위신간질림파세포침윤급간질섬유화，B조최위현저。정약기말，A、B조혈청뇨소담수평분별위(17.96±2.04)、(19.20±3.36) mmol/L，균고우C조적（8.30±1.79）mmol/L(균P＜0.01)；B조기항수평(24.20±5.28) mmol/L고우C조(9.87 ±2.71) mmol/L(P ＜0.01)，차분별교용약기수평승고（분별위P＜0.01,P＜0.05）；량실험조신장계수교전하강；신장잉유결정미배출，간질개변교정약전미견감경。결론(1)사위3주령강단유웅성Wistar대서함삼취청알질량분수위2％적사료15주능구건립삼취청알비뇨계결석모형。사위함삼취청알질량분수위1％적사료15주，부소수대서가형성육안가견적신장결석，부분형성경하결정。결석성분주요위삼취청알(＞90％)、소량뇨산급겁소량삼취청산。(2)삼취청알가도치대서신공능수손，경하가견신소관결정형성、신소관확장、염증、섬유화등병리표현，병구유제량의뢰성。삼취청알적신장손해여결정、결석경조유일정상관성。(3)정지섭입삼취청알후，신공능급신장염증、섬유화등개변단기내미견호전，결정자행배출적과정교위완만，여결정지속존재가능회조성진일보적혹불가역전적신장손상。
ObjectiveTo investigate melamine-induced pathological changes in the kidney. MethodWistar rats were fed with a diet containing 0, 1％ and 2％ melamine for 15 weeks. After melamine feeding was stopped, various outcome measures were observed for 4 weeks. Result Rats fed with melamine showed reduced caloric intake, slower weight gain and impaired renal function. The blood urea nitrogen of group A and B [ ( 13.23 ± 5. 10 ) mmol/L and ( 18.30 ± 5.90 ) mmol/L, respectively ] and serum creatinine levels of group B [ ( 19.90 ± 2.90 ) mmol/L ] were higher than that of group C [ ( 8.23 ±2.30) mmol/L and ( 10. 04 ± 1. 73) mmol/L] (P ＜0. 01, respectively). Additionally, the kidney coefficients of group A and B were higher than that of group C (P ＜ 0. 01, respectively). Crystals, tubular ectasia and interstitial inflammation and fibrosis were found in the kidneys of melamine fed rats. Four weeks after discontinuation of feeding with melamine-contained diet, the caloric intake and weight of the rats increased, the coefficients of the kidney decreased, and the blood urea nitrogen of group A and B [ ( 17.96 ± 2.04) mmol/L and ( 19.20 ± 3.36) mmol/L, respectively] and serum creatinine levels of group B [(24. 20 ±5. 28) mmoL/L], which became worse than 4 weeks before (P ＜0. 01;P ＜0.05,respectively), and were still higher than that of group C [ ( 8.30 ± 1.79) mmol/L and ( 9.87 ± 2.71 )mmol/L, P ＜0.01, respectively]. Crystals remained inside the kidney, changes in the renal interstitium did not improve. Conclusion( 1 ) Melamine-induced urinary calculus rat model can be established by feeding 3-week old male Wistar rats with a diet containing 2％ melamine for 15 weeks. The main constituent of the urinary calculus was melamine ( ＞ 90％ ), with a little uric acid and traces of cyanuric acid. (2)Melamine damaged the renal function, formed renal crystals, and led to the pathological changes of kidneys. All the influences seemed to be dose-depended and was related with the obstruction of the crystals or caculus in the kidney. (3)The renal function and the pathological changes did not improve 4 weeks after discontinuation of feeding with melamine-contained diet.