中国老年学杂志
中國老年學雜誌
중국노년학잡지
CHINESE JOURNAL OF GERONTOLOGY
2014年
10期
2770-2773
,共4页
张涛%迟雁青%李亚%刘茂东%李英
張濤%遲雁青%李亞%劉茂東%李英
장도%지안청%리아%류무동%리영
糖尿病肾病%抗增殖因子%凋亡诱导因子%凋亡%肾小管上皮细胞%别嘌醇
糖尿病腎病%抗增殖因子%凋亡誘導因子%凋亡%腎小管上皮細胞%彆嘌醇
당뇨병신병%항증식인자%조망유도인자%조망%신소관상피세포%별표순
Diabetic nephropathy%Prohibitin%Apoptosis inducing factor%Apoptosis%Renal tubular epithelia cell%Allopurinol
目的：探讨抑制尿酸药物别嘌醇对2型糖尿病db/db小鼠肾小管上皮细胞prohibitin表达及细胞凋亡的影响。方法将 T2 DM db/db 小鼠随机分为：db/db 小鼠对照组（ db/db组）、db/db小鼠＋别嘌醇50 mg · kg-1· d-1治疗组（ db/db ＋A 组）；将 db/m 小鼠作为对照组（ db/m组），干预12 w后检测血糖、血尿素氮、血尿酸、血甘油三酯、24 h尿蛋白定量水平；电镜下观察肾小管上皮细胞病理改变；Western 印迹及免疫组化法检测肾皮质抗增殖因子（prohibitin）、凋亡诱导因子（AIF）的定位与表达水平；分光光度计检测肾皮质丙二醛（MDA）的含量；原位末端脱氧核糖核酸转移酶介导的脱氧尿苷三磷酸标记法（ TUNEL法）检测肾小管间质细胞凋亡情况。结果①12 w 末，db/db组小鼠血糖、血尿酸、血甘油三酯、血尿素氮、24 h尿蛋白定量、肾组织匀浆MDA含量较db/m组均有不同程度的升高（ P＜0.05），而db/db＋A组血尿酸、血尿素氮、24 h尿蛋白定量、肾组织MDA含量均较同时期db/db组降低（ P＜0.05）。②免疫组化与Western印迹检测结果均提示，db/db组肾皮质 prohibitin蛋白表达量较db/m组明显降低（P＜0.01），db/db＋A组表达比db/db组有所增强（P＜0.01）。 db/db组肾皮质AIF蛋白表达明显高于 db/m 组（P＜0.01），db/db＋A 组表达比db/db组有所降低（P＜0.01）。③电镜下db/db组小鼠肾小管上皮细胞线粒体可见肿胀、空泡变，嵴结构消失等病变，db/db＋A组上述病变减轻。④TUNEL检测结果：db/db组小鼠肾小管上皮细胞及间质细胞凋亡数量较db/m组明显增多，db/db＋A组肾小管上皮细胞凋亡率较 db/db组降低。结论别嘌醇可抑制T2DM小鼠肾小管间质细胞凋亡，其机制可能与其降低血尿酸水平后提高肾组织 prohibitin的表达，减少局部氧化应激反应，抑制AIF增多并向核内转位有关。
目的：探討抑製尿痠藥物彆嘌醇對2型糖尿病db/db小鼠腎小管上皮細胞prohibitin錶達及細胞凋亡的影響。方法將 T2 DM db/db 小鼠隨機分為：db/db 小鼠對照組（ db/db組）、db/db小鼠＋彆嘌醇50 mg · kg-1· d-1治療組（ db/db ＋A 組）；將 db/m 小鼠作為對照組（ db/m組），榦預12 w後檢測血糖、血尿素氮、血尿痠、血甘油三酯、24 h尿蛋白定量水平；電鏡下觀察腎小管上皮細胞病理改變；Western 印跡及免疫組化法檢測腎皮質抗增殖因子（prohibitin）、凋亡誘導因子（AIF）的定位與錶達水平；分光光度計檢測腎皮質丙二醛（MDA）的含量；原位末耑脫氧覈糖覈痠轉移酶介導的脫氧尿苷三燐痠標記法（ TUNEL法）檢測腎小管間質細胞凋亡情況。結果①12 w 末，db/db組小鼠血糖、血尿痠、血甘油三酯、血尿素氮、24 h尿蛋白定量、腎組織勻漿MDA含量較db/m組均有不同程度的升高（ P＜0.05），而db/db＋A組血尿痠、血尿素氮、24 h尿蛋白定量、腎組織MDA含量均較同時期db/db組降低（ P＜0.05）。②免疫組化與Western印跡檢測結果均提示，db/db組腎皮質 prohibitin蛋白錶達量較db/m組明顯降低（P＜0.01），db/db＋A組錶達比db/db組有所增彊（P＜0.01）。 db/db組腎皮質AIF蛋白錶達明顯高于 db/m 組（P＜0.01），db/db＋A 組錶達比db/db組有所降低（P＜0.01）。③電鏡下db/db組小鼠腎小管上皮細胞線粒體可見腫脹、空泡變，嵴結構消失等病變，db/db＋A組上述病變減輕。④TUNEL檢測結果：db/db組小鼠腎小管上皮細胞及間質細胞凋亡數量較db/m組明顯增多，db/db＋A組腎小管上皮細胞凋亡率較 db/db組降低。結論彆嘌醇可抑製T2DM小鼠腎小管間質細胞凋亡，其機製可能與其降低血尿痠水平後提高腎組織 prohibitin的錶達，減少跼部氧化應激反應，抑製AIF增多併嚮覈內轉位有關。
목적：탐토억제뇨산약물별표순대2형당뇨병db/db소서신소관상피세포prohibitin표체급세포조망적영향。방법장 T2 DM db/db 소서수궤분위：db/db 소서대조조（ db/db조）、db/db소서＋별표순50 mg · kg-1· d-1치료조（ db/db ＋A 조）；장 db/m 소서작위대조조（ db/m조），간예12 w후검측혈당、혈뇨소담、혈뇨산、혈감유삼지、24 h뇨단백정량수평；전경하관찰신소관상피세포병리개변；Western 인적급면역조화법검측신피질항증식인자（prohibitin）、조망유도인자（AIF）적정위여표체수평；분광광도계검측신피질병이철（MDA）적함량；원위말단탈양핵당핵산전이매개도적탈양뇨감삼린산표기법（ TUNEL법）검측신소관간질세포조망정황。결과①12 w 말，db/db조소서혈당、혈뇨산、혈감유삼지、혈뇨소담、24 h뇨단백정량、신조직균장MDA함량교db/m조균유불동정도적승고（ P＜0.05），이db/db＋A조혈뇨산、혈뇨소담、24 h뇨단백정량、신조직MDA함량균교동시기db/db조강저（ P＜0.05）。②면역조화여Western인적검측결과균제시，db/db조신피질 prohibitin단백표체량교db/m조명현강저（P＜0.01），db/db＋A조표체비db/db조유소증강（P＜0.01）。 db/db조신피질AIF단백표체명현고우 db/m 조（P＜0.01），db/db＋A 조표체비db/db조유소강저（P＜0.01）。③전경하db/db조소서신소관상피세포선립체가견종창、공포변，척결구소실등병변，db/db＋A조상술병변감경。④TUNEL검측결과：db/db조소서신소관상피세포급간질세포조망수량교db/m조명현증다，db/db＋A조신소관상피세포조망솔교 db/db조강저。결론별표순가억제T2DM소서신소관간질세포조망，기궤제가능여기강저혈뇨산수평후제고신조직 prohibitin적표체，감소국부양화응격반응，억제AIF증다병향핵내전위유관。
Objective To observe the effect of allopurinol on prohibitin expression and cell apoptosis of renal tubulointerstitium in type 2 diabetic mice and investigate the possible mechanism .Methods 18 mice were divided into db/m ( normal contral ) ,db/db ( diabetic contral), db/db+A (db/db mice treated with allopurinol 50 mg· kg-1· d-1)groups.After treatment for 12 weeks, blood glucose, uric acid, urea nitrogen, creatinine, triglyceride, proteinuria and level of MDA in the renal cortex were measured .Tubulointersititial pathology injury were observed by electron microscope .The protein location and expressions of prohibitin ,AIF in kidney were observed by immunohisto-chemistry and Western blot .Tubulointersititial cell apoptosis was assessed by TUNEL .Results Compared with those of db/m group, the levels of blood glucose, uric acid, urea nitrogen, triglyceride, proteinuria and cortical MDA content were increased in the DM models .The parameters such as blood uric acid , urea nitrogen , proteinuria and cortical MDA content were remarkably improved with allopurinol interven -tion.The protein expression of prohibitin in the kidney of db/db group was reduced obviously than that of db /m group, while the expression of AIF was increased .The changes of above two parameters were improved in allopurinol intervention group .TUNEL results showed that cell apoptosis of renal tubulointerstitium in db/db group was obvious ,while the apoptosis rate was down-regulated in db/db+A group than that of db/db group .Conclusions Allopurinol treatment could significantly inhibit the cell apoptosis of renal tubulointerstitium in type 2 diabetic mice,which maybe through the mechanism of reducing the blood uric acid level ,and increasing prohibitin expression in renal tissue , decrea-sing the local oxidative stress and inhibiting AIF expression and its nuclear translocation .
