中华糖尿病杂志
中華糖尿病雜誌
중화당뇨병잡지
CHINES JOURNAL OF DLABETES MELLITUS
2015年
4期
245-249
,共5页
陆晓兰%杨慧%刘文娟%王蒙%杨叶萍%严琳玲%谢丽娜%王熠%鹿斌%胡吉%李益明%胡仁明%张朝云
陸曉蘭%楊慧%劉文娟%王矇%楊葉萍%嚴琳玲%謝麗娜%王熠%鹿斌%鬍吉%李益明%鬍仁明%張朝雲
륙효란%양혜%류문연%왕몽%양협평%엄림령%사려나%왕습%록빈%호길%리익명%호인명%장조운
糖尿病肾病%螺内酯%db/db小鼠
糖尿病腎病%螺內酯%db/db小鼠
당뇨병신병%라내지%db/db소서
Diabetic nephropathies%Spironolactone%db/db mice
目的：利用db/db 2型糖尿病小鼠模型，验证螺内酯的肾脏保护作用并探讨其机制。方法将16只db/db小鼠随机数字表法分为螺内酯干预组（DM-SPR组，n=8）和糖尿病对照组（DM组， n=8），同窝出生的db/m小鼠（NC组，n=10）作为正常对照组。小鼠6周龄时，DM-SPR组予20 mg/（kg· d）螺内酯干预，DM组和NC组予生理盐水干预，期间监测小鼠的血糖、血压、尿白蛋白/肌酐比值，干预12周后处死小鼠，留取肾脏组织。应用透射电镜观察肾脏超微结构。应用实时聚合酶链反应（RT-PCR）检测炎症相关因子单核细胞趋化蛋白（MCP）-1、白细胞介素（IL）-1β、CD68和纤维化指标胶原Ⅳ、纤连蛋白、转化生长因子（TGF）-β1。采用单因素方差分析和q检验进行组间数据比较。结果18周龄时，DM组和DM-SPR组小鼠血糖、糖化血红蛋白（HbA1c）均高于NC组（血糖：q=-12.09、-14.49，HbA1c：q=-8.86、-11.90，均P<0.01），而DM组和DM-SPR组小鼠血糖及HbA1c无显著差异（q=-2.17、-2.09，均P>0.05）。三组小鼠收缩压和舒张压差异均无统计学意义（F=2.05、2.75，均P>0.05）。三组尿白蛋白肌酐比差异有统计学意义（F=9.12，P<0.05），而DM组显著高于NC组（q=-5.79，P<0.01），DM-SPR组显著低于DM组（q=4.03，P<0.05）。与NC组小鼠相比，电镜下DM组小鼠肾脏内皮细胞窗孔显著减少，基底膜增厚，足突广泛融合，而DM-SPR组有效上述情况明显改善。RT-PCR显示3组CD68、MCP-1、IL-1β和TGF-β1、胶原Ⅳ、纤连蛋白表达差异均有统计学意义（F=4.38～9.85，均P<0.05）；与NC组比较，DM组以上因子表达均显著升高（q=-6.18～-3.53，均P<0.05），而DM-SPR组以上因子的表达较DM组均显著降低（q=0.23～4.64，均P<0.05）。结论螺内酯对2型糖尿病小鼠肾脏具有保护作用，该作用不依赖于降糖和降血压，可能与其拮抗炎症因子、抗纤维化有关。
目的：利用db/db 2型糖尿病小鼠模型，驗證螺內酯的腎髒保護作用併探討其機製。方法將16隻db/db小鼠隨機數字錶法分為螺內酯榦預組（DM-SPR組，n=8）和糖尿病對照組（DM組， n=8），同窩齣生的db/m小鼠（NC組，n=10）作為正常對照組。小鼠6週齡時，DM-SPR組予20 mg/（kg· d）螺內酯榦預，DM組和NC組予生理鹽水榦預，期間鑑測小鼠的血糖、血壓、尿白蛋白/肌酐比值，榦預12週後處死小鼠，留取腎髒組織。應用透射電鏡觀察腎髒超微結構。應用實時聚閤酶鏈反應（RT-PCR）檢測炎癥相關因子單覈細胞趨化蛋白（MCP）-1、白細胞介素（IL）-1β、CD68和纖維化指標膠原Ⅳ、纖連蛋白、轉化生長因子（TGF）-β1。採用單因素方差分析和q檢驗進行組間數據比較。結果18週齡時，DM組和DM-SPR組小鼠血糖、糖化血紅蛋白（HbA1c）均高于NC組（血糖：q=-12.09、-14.49，HbA1c：q=-8.86、-11.90，均P<0.01），而DM組和DM-SPR組小鼠血糖及HbA1c無顯著差異（q=-2.17、-2.09，均P>0.05）。三組小鼠收縮壓和舒張壓差異均無統計學意義（F=2.05、2.75，均P>0.05）。三組尿白蛋白肌酐比差異有統計學意義（F=9.12，P<0.05），而DM組顯著高于NC組（q=-5.79，P<0.01），DM-SPR組顯著低于DM組（q=4.03，P<0.05）。與NC組小鼠相比，電鏡下DM組小鼠腎髒內皮細胞窗孔顯著減少，基底膜增厚，足突廣汎融閤，而DM-SPR組有效上述情況明顯改善。RT-PCR顯示3組CD68、MCP-1、IL-1β和TGF-β1、膠原Ⅳ、纖連蛋白錶達差異均有統計學意義（F=4.38～9.85，均P<0.05）；與NC組比較，DM組以上因子錶達均顯著升高（q=-6.18～-3.53，均P<0.05），而DM-SPR組以上因子的錶達較DM組均顯著降低（q=0.23～4.64，均P<0.05）。結論螺內酯對2型糖尿病小鼠腎髒具有保護作用，該作用不依賴于降糖和降血壓，可能與其拮抗炎癥因子、抗纖維化有關。
목적：이용db/db 2형당뇨병소서모형，험증라내지적신장보호작용병탐토기궤제。방법장16지db/db소서수궤수자표법분위라내지간예조（DM-SPR조，n=8）화당뇨병대조조（DM조， n=8），동와출생적db/m소서（NC조，n=10）작위정상대조조。소서6주령시，DM-SPR조여20 mg/（kg· d）라내지간예，DM조화NC조여생리염수간예，기간감측소서적혈당、혈압、뇨백단백/기항비치，간예12주후처사소서，류취신장조직。응용투사전경관찰신장초미결구。응용실시취합매련반응（RT-PCR）검측염증상관인자단핵세포추화단백（MCP）-1、백세포개소（IL）-1β、CD68화섬유화지표효원Ⅳ、섬련단백、전화생장인자（TGF）-β1。채용단인소방차분석화q검험진행조간수거비교。결과18주령시，DM조화DM-SPR조소서혈당、당화혈홍단백（HbA1c）균고우NC조（혈당：q=-12.09、-14.49，HbA1c：q=-8.86、-11.90，균P<0.01），이DM조화DM-SPR조소서혈당급HbA1c무현저차이（q=-2.17、-2.09，균P>0.05）。삼조소서수축압화서장압차이균무통계학의의（F=2.05、2.75，균P>0.05）。삼조뇨백단백기항비차이유통계학의의（F=9.12，P<0.05），이DM조현저고우NC조（q=-5.79，P<0.01），DM-SPR조현저저우DM조（q=4.03，P<0.05）。여NC조소서상비，전경하DM조소서신장내피세포창공현저감소，기저막증후，족돌엄범융합，이DM-SPR조유효상술정황명현개선。RT-PCR현시3조CD68、MCP-1、IL-1β화TGF-β1、효원Ⅳ、섬련단백표체차이균유통계학의의（F=4.38～9.85，균P<0.05）；여NC조비교，DM조이상인자표체균현저승고（q=-6.18～-3.53，균P<0.05），이DM-SPR조이상인자적표체교DM조균현저강저（q=0.23～4.64，균P<0.05）。결론라내지대2형당뇨병소서신장구유보호작용，해작용불의뢰우강당화강혈압，가능여기길항염증인자、항섬유화유관。
Objective To investigate the renal protective effects of spironolactone(SPR)and its possible mechanism indiabetic db/db mice. Methods Sixteen diabetic db/db mice were randomly divided into two groups by random number table:DM-SPR group(n=8)and DM group(n=8). Littermate db/m mice (NC group, n=10) were usedas normal control.Six-week-old DM-SPR mice were treated with SPR [20 mg/(kg·d)], while DM and NC group were both treated with nomal saline(NS) for 12 weeks.During the experiment, blood glucose, blood pressure and urinary albumin/creatine were measured. After the treatment for 12 weeks, all the mice were sacrificed and thekidneys were collected. Glomerular ultrastructure were observed undertransmission electron microscope. The mRNA expression of inflammation cytokines monocyte chemotactic protein-1(MCP-1), interleukin(IL)-1β,CD68 and fibrosis index collagen Ⅳ, fibronectin, transforming growth factor-β(TGF-β) were detected by realtime polymerase chain reaction(RT-PCR). One-way analysisofvarianceand q test were used for dataanalysis among groups. Results At the 18-week, the levels of blood glucose and glycated hemoglobin A1c(HbA1c) were both significantly higher in DM groupand DM-SPR groupthan those in NC group(glucose:q=-12.09,-14.49;HbA1c:q=-8.86,-11.90, all P<0.05),and there was no significant difference between DM group and DM-SPR group(q=-2.17,-2.09, all P>0.05).There was no significant differences in systolic and diastolic pressure among the three groups(F=2.05, 2.75, bothP>0.05).There was significant differences in urinary albumin/creatine among the three groups(F=9.12,P<0.05), it was higher in the DM group than that in the NC group(q=-5.79, P<0.05),and it was lower in the DM-SPR group than that in the DM group(q=4.03, P<0.05). In the DM group, reduction of fenestrated endothelium, thickening of glomerular basement memebrane(GBM)and diffuse effacement of podocyte foot process were commonly found underelectron microscope, while SPR treatment effectively improved theseinjuries.RT-PCR analysis showed that there were significant differences in mRNA expression of inflammatory cytokines CD68,MCP-1,IL-1βand fibrosis indexes TGF-β1, collagenⅣ, fibronectin among the three groups(F=4.38 to 9.85, all P<0.05);the expression of above mentioned indexes were all elevated in the DM groupwhen compared with those in the NC group(q=-6.18 to-3.53, all P<0.05), and the SPR treatment effectively decreased the expression of those indexes when compared with those in DM group(q=0.23 to 4.64, all P<0.05).Conclusion SPRplays protective rolesin the diabetic nephropathy in db/db mice, which maybe mediated by attenuatinginflammation and fibrosis.
