糖尿病肾病大鼠骨骼肌蛋白消耗及低蛋白联合α酮酸的作用
당뇨병신병대서골격기단백소모급저단백연합α동산적작용
Skeletal muscle wasting in diabetic kidney disease rats and the effect of low-protein diet combined with a-keto acids
  • 万方数据
    中华肾脏病杂志 중화신장병잡지

    2013年 3期 204-209 ,共6页

    王嘉琳%谷立杰%袁伟杰%黄娟%董婷%唐知还

    왕가림%곡립걸%원위걸%황연%동정%당지환

    糖尿病肾病%肌,骨骼%能量代谢%α酮酸%低蛋白饮食 糖尿病腎病%肌,骨骼%能量代謝%α酮痠%低蛋白飲食
    당뇨병신병%기,골격%능량대사%α동산%저단백음식
    Diabetic nephropathies%Skeletal muscle%Energy metabolism%α keto acid%Low-protein diet
    目的 观察Goto-Kakizaki(GK)大鼠2型非肥胖糖尿病肾病(DKD)模型骨骼肌蛋白消耗情况,探讨低蛋白联合α酮酸对其的作用.方法 雄性GK大鼠45只,24周龄开始给予饮食干预,随机分为正常蛋白组(22％酪蛋白,NPD组)、低蛋白组(6％酪蛋白,LPD组)和低蛋白联合α酮酸组(5％酪蛋白+l％α酮酸,Keto组)；另设性别、周龄相同的Wistar大鼠15只为对照组(CTL组),予以正常蛋白饮食(22％酪蛋白).观察大鼠生存状况,每周用电子秤称重,同时检测各组第24、32、40、48周龄大鼠尿总蛋白及尿白蛋白,血清中葡萄糖、Scr、BUN、白蛋白、胆固醇、三酰甘油等变化.组织病理学观察48周龄大鼠比目鱼肌形态,并计算肌纤维横截面积.用定量PCR和Western印迹法检测比目鱼肌肌萎缩关键基因atrogin-1、MuRF-1和骨骼肌增殖分化关键基因MyoD、myogenin表达水平.结果 与CTL组相比,NPD、LPD及Keto组体质量显著下降[(317.90±13.81)、(330.38±11.96)、(390.44±12.25)g比(429.43±16.85)g,均P＜0.05]；尿白蛋白排泄增多[(14.36±5.52)、(8.12±4.61)、(5.58±3.50) mg/24 h比(0.61±0.16) mg/24 h,均P＜0.05]；Scr和BUN水平也显著增高[Scr:(81.50±7.88)、(66.32±8.36)、(63.44±8.21) μmol/L比(24.43±6.15) μmol/L; BUN:(7.53±1.05)、(5.63±1.40)、(5.54±0.97) mmol/L比(2.98±0.62) mmol/L；均P＜0.05].与CTL组相比,NPD、LPD及Keto组比目鱼肌变细,肌纤维横截面积减少(均P＜ 0.05)；肌肉组织atrogin-1和MuRF-1表达水平约为CTL组的2倍(均P＜0.05),而MyoD和myogenin表达水平均有不同程度下降(均P＜0.05).与NPD、LPD组相比,Keto组GK大鼠40周龄时体质量显著增加[(381.62±15.82)g比(331.50±17.58)、(326.60±13.43)g,均P＜0.05]；Scr、BUN及尿蛋白水平均显著降低(均P＜0.05)；比目鱼肌湿重和横截面积轻度增加；atrogin-1、MuRF-1蛋白表达水平显著下降；myogenin、MyoD蛋白表达水平高于CTL组(均P＜0.05).LPD组和NPD组组间差异无统计学意义.结论 DKD可引起骨骼肌蛋白质消耗；肌萎缩关键基因表达升高；骨骼肌卫星细胞的增殖分化功能障碍.低蛋白联合α酮酸饮食则可改善DKD肌肉蛋白消耗状态,减轻骨骼肌萎缩程度.
    목적 관찰Goto-Kakizaki(GK)대서2형비비반당뇨병신병(DKD)모형골격기단백소모정황,탐토저단백연합α동산대기적작용.방법 웅성GK대서45지,24주령개시급여음식간예,수궤분위정상단백조(22％락단백,NPD조)、저단백조(6％락단백,LPD조)화저단백연합α동산조(5％락단백+l％α동산,Keto조)；령설성별、주령상동적Wistar대서15지위대조조(CTL조),여이정상단백음식(22％락단백).관찰대서생존상황,매주용전자칭칭중,동시검측각조제24、32、40、48주령대서뇨총단백급뇨백단백,혈청중포도당、Scr、BUN、백단백、담고순、삼선감유등변화.조직병이학관찰48주령대서비목어기형태,병계산기섬유횡절면적.용정량PCR화Western인적법검측비목어기기위축관건기인atrogin-1、MuRF-1화골격기증식분화관건기인MyoD、myogenin표체수평.결과 여CTL조상비,NPD、LPD급Keto조체질량현저하강[(317.90±13.81)、(330.38±11.96)、(390.44±12.25)g비(429.43±16.85)g,균P＜0.05]；뇨백단백배설증다[(14.36±5.52)、(8.12±4.61)、(5.58±3.50) mg/24 h비(0.61±0.16) mg/24 h,균P＜0.05]；Scr화BUN수평야현저증고[Scr:(81.50±7.88)、(66.32±8.36)、(63.44±8.21) μmol/L비(24.43±6.15) μmol/L; BUN:(7.53±1.05)、(5.63±1.40)、(5.54±0.97) mmol/L비(2.98±0.62) mmol/L；균P＜0.05].여CTL조상비,NPD、LPD급Keto조비목어기변세,기섬유횡절면적감소(균P＜ 0.05)；기육조직atrogin-1화MuRF-1표체수평약위CTL조적2배(균P＜0.05),이MyoD화myogenin표체수평균유불동정도하강(균P＜0.05).여NPD、LPD조상비,Keto조GK대서40주령시체질량현저증가[(381.62±15.82)g비(331.50±17.58)、(326.60±13.43)g,균P＜0.05]；Scr、BUN급뇨단백수평균현저강저(균P＜0.05)；비목어기습중화횡절면적경도증가；atrogin-1、MuRF-1단백표체수평현저하강；myogenin、MyoD단백표체수평고우CTL조(균P＜0.05).LPD조화NPD조조간차이무통계학의의.결론 DKD가인기골격기단백질소모；기위축관건기인표체승고；골격기위성세포적증식분화공능장애.저단백연합α동산음식칙가개선DKD기육단백소모상태,감경골격기위축정도.
    Objective To observe the muscle wasting in diabetic kidney disease (DKD) model of type 2 and non-obese diabetes mellitus in Goto-Kakizaki (GK) rats,and to evaluate the effect of lowprotein diet supplemented with α-keto acids on muscle wasting.Methods Forty-five male 24-weekage GK rats were randomly divided into three groups:normal protein diet group (22％ casein diet,NPD),low protein diet group (6％ casein diet,LPD) and LPD + α-keto group (5％ casein + 1％ α-keto,Keto).Fifteen gender-and age-matched Wistar rats were served as the control group (CTL).The living condition of GK rats was observed and body weight was measured once a week.Urine albumin,serum glucose,lipids,albumin,creatinine and urea nitrogen were measured at the age of 24,32,40,48 weeks.Soleus muscle at the age of 48-week was observed to calculate the muscle size with software.Expressions of atrogin-1,MuRF-1 and MyoD,myogenin were examined by Q-PCR and Western blotting.Results Compared with the CTL group,NPD,LPD,Keto groups had lower body weight [(317.90± 13.81),(330.38±11.96),(390.44±12.25) g vs (429.43± 16.85) g,all P ＜ 0.05],higher urine albumin [(14.36±5.52),(8.12±4.61),(5.58±3.50) mg/24 h vs (0.61±0.16) mg/24 h,all P ＜ 0.05],higher serum creatinine [(81.50±7.88),(66.32±8.36),(63.44±8.21) μmol/L vs (24.43±6.15) μmol/L,all P ＜0.05] and urea nitrogen [(7.53±1.05),(5.63±1.40),(5.54±0.97) mmol/L vs (2.98±0.62) mmol/L,all P ＜0.05].The cross-sectional area of soleus muscle fibers was larger in CTL group.Compared with CTL group,the expression levels of atrogin-1 and MuRF-1 increased significantly (all P ＜ 0.05),and of MyoD and myogenin decreased significantly in NPD,LPD,Keto groups (all P ＜ 0.05).In Keto group after 40 weeks,muscle wasting was improved compared with NPD and LPD group [body weight (381.62± 15.82) g vs (331.50±17.58),(326.60± 13.43) g,all P ＜ 0.05],cross-sectional area of soleus muscle increased,levels of urine albumin,serum creatinine and urea nitrogen decreased (all P ＜ 0.05),the protein expressions of atrogin-1 and MuRF-1 decreased,and myogenin and MyoD were higher as compared to CTL group (all P ＜ 0.05).There were no significant differences between NPD and LPD group.Conclusions In DKD condition,protein degradation in the skeletal muscle is accelerated,the genes which control muscle atrophy are activated,and proliferation and differentiation of the muscle satellite cells are impaired.Low-protein diet supplemented with α-keto acids can improve muscle wasting induced by DKD.
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