中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2013年
11期
824-829
,共6页
王嘉琳%袁伟杰%谷立杰%黄娟%董婷
王嘉琳%袁偉傑%穀立傑%黃娟%董婷
왕가림%원위걸%곡립걸%황연%동정
糖尿病肾病%肌,骨骼%能量代谢%线粒体%膳食,限制蛋白质
糖尿病腎病%肌,骨骼%能量代謝%線粒體%膳食,限製蛋白質
당뇨병신병%기,골격%능량대사%선립체%선식,한제단백질
Diabetic nephropathies%Muscle,skeletal%Energy metabolism%Mitochondrion%Diet,protein-restricted
目的 观察糖尿病肾病(DKD)动物模型Goto-Kakizaki(GK)大鼠骨骼肌蛋白消耗相关线粒体损伤情况,及低蛋白联合α酮酸对其作用.方法 45只24周龄雄性GK大鼠随机分入正常蛋白组(NPD组)、低蛋白组(LPD组)和低蛋白联合α酮酸组(Keto组);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.每周称重,同时第24、32、40、48周时检测大鼠尿总蛋白及尿白蛋白、血清葡萄糖、Scr、BUN等变化.光镜下观察48周龄大鼠比目鱼肌琥珀酸脱氢酶(SDH)、烟酰胺腺嘌呤二核苷酸四唑氧化还原酶(NADH)染色后的形态、酶活性,计算肌纤维横截面积和Ⅰ、Ⅱ型肌纤维比例.透射电镜下观察组织超微结构,用分光光度计法检测比目鱼肌柠檬酸合成酶活性,定量PCR法检测线粒体DNA表达.结果 与CTL组相比,NPD、LPD及Keto组体质量显著下降,尿白蛋白排泄增多,Scr和BUN水平也显著增高;肌纤维横截面积减少(均P＜ 0.05),Ⅱ型肌纤维比例增加[(37.01±1.85)％、(38.72±1.67)％、(26.77±2.23)％比(18.65±2.37)％,均P＜0.05];电镜下肌丝部分断裂,线粒体肿胀变形明显;NPD及LPD组肌肉组织柠檬酸合成酶活性降低[(19 260.83±3522.13)、(21 313.11±2266.89) U· min-1·g1比(24 787.47±1833.76) U· min-1·g-1,均P＜ 0.05],线粒体DNA数量减少.与NPD、LPD组相比,Keto组GK大鼠体质量增加显著,血清Scr、BUN及尿蛋白水平均显著降低;比目鱼肌湿重和横截面积轻度增加,Ⅱ型肌纤维比例降低,电镜下肌丝完整,线粒体形态趋于正常,柠檬酸合成酶活性及线粒体DNA表达显著增加(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,伴线粒体肿胀变形,DNA表达减少及氧化磷酸化功能障碍.低蛋白联合α酮酸饮食则可改善DKD骨骼肌线粒体损伤,缓解蛋白质消耗状况,减轻骨骼肌萎缩程度.
目的 觀察糖尿病腎病(DKD)動物模型Goto-Kakizaki(GK)大鼠骨骼肌蛋白消耗相關線粒體損傷情況,及低蛋白聯閤α酮痠對其作用.方法 45隻24週齡雄性GK大鼠隨機分入正常蛋白組(NPD組)、低蛋白組(LPD組)和低蛋白聯閤α酮痠組(Keto組);性彆、週齡相同的Wistar大鼠15隻為對照組(CTL組),予以正常蛋白飲食.每週稱重,同時第24、32、40、48週時檢測大鼠尿總蛋白及尿白蛋白、血清葡萄糖、Scr、BUN等變化.光鏡下觀察48週齡大鼠比目魚肌琥珀痠脫氫酶(SDH)、煙酰胺腺嘌呤二覈苷痠四唑氧化還原酶(NADH)染色後的形態、酶活性,計算肌纖維橫截麵積和Ⅰ、Ⅱ型肌纖維比例.透射電鏡下觀察組織超微結構,用分光光度計法檢測比目魚肌檸檬痠閤成酶活性,定量PCR法檢測線粒體DNA錶達.結果 與CTL組相比,NPD、LPD及Keto組體質量顯著下降,尿白蛋白排洩增多,Scr和BUN水平也顯著增高;肌纖維橫截麵積減少(均P＜ 0.05),Ⅱ型肌纖維比例增加[(37.01±1.85)％、(38.72±1.67)％、(26.77±2.23)％比(18.65±2.37)％,均P＜0.05];電鏡下肌絲部分斷裂,線粒體腫脹變形明顯;NPD及LPD組肌肉組織檸檬痠閤成酶活性降低[(19 260.83±3522.13)、(21 313.11±2266.89) U· min-1·g1比(24 787.47±1833.76) U· min-1·g-1,均P＜ 0.05],線粒體DNA數量減少.與NPD、LPD組相比,Keto組GK大鼠體質量增加顯著,血清Scr、BUN及尿蛋白水平均顯著降低;比目魚肌濕重和橫截麵積輕度增加,Ⅱ型肌纖維比例降低,電鏡下肌絲完整,線粒體形態趨于正常,檸檬痠閤成酶活性及線粒體DNA錶達顯著增加(均P＜0.05).LPD組和NPD組組間差異無統計學意義.結論 DKD可引起骨骼肌蛋白質消耗,伴線粒體腫脹變形,DNA錶達減少及氧化燐痠化功能障礙.低蛋白聯閤α酮痠飲食則可改善DKD骨骼肌線粒體損傷,緩解蛋白質消耗狀況,減輕骨骼肌萎縮程度.
목적 관찰당뇨병신병(DKD)동물모형Goto-Kakizaki(GK)대서골격기단백소모상관선립체손상정황,급저단백연합α동산대기작용.방법 45지24주령웅성GK대서수궤분입정상단백조(NPD조)、저단백조(LPD조)화저단백연합α동산조(Keto조);성별、주령상동적Wistar대서15지위대조조(CTL조),여이정상단백음식.매주칭중,동시제24、32、40、48주시검측대서뇨총단백급뇨백단백、혈청포도당、Scr、BUN등변화.광경하관찰48주령대서비목어기호박산탈경매(SDH)、연선알선표령이핵감산사서양화환원매(NADH)염색후적형태、매활성,계산기섬유횡절면적화Ⅰ、Ⅱ형기섬유비례.투사전경하관찰조직초미결구,용분광광도계법검측비목어기저몽산합성매활성,정량PCR법검측선립체DNA표체.결과 여CTL조상비,NPD、LPD급Keto조체질량현저하강,뇨백단백배설증다,Scr화BUN수평야현저증고;기섬유횡절면적감소(균P＜ 0.05),Ⅱ형기섬유비례증가[(37.01±1.85)％、(38.72±1.67)％、(26.77±2.23)％비(18.65±2.37)％,균P＜0.05];전경하기사부분단렬,선립체종창변형명현;NPD급LPD조기육조직저몽산합성매활성강저[(19 260.83±3522.13)、(21 313.11±2266.89) U· min-1·g1비(24 787.47±1833.76) U· min-1·g-1,균P＜ 0.05],선립체DNA수량감소.여NPD、LPD조상비,Keto조GK대서체질량증가현저,혈청Scr、BUN급뇨단백수평균현저강저;비목어기습중화횡절면적경도증가,Ⅱ형기섬유비례강저,전경하기사완정,선립체형태추우정상,저몽산합성매활성급선립체DNA표체현저증가(균P＜0.05).LPD조화NPD조조간차이무통계학의의.결론 DKD가인기골격기단백질소모,반선립체종창변형,DNA표체감소급양화린산화공능장애.저단백연합α동산음식칙가개선DKD골격기선립체손상,완해단백질소모상황,감경골격기위축정도.
Objective To observe the mitochondrial damage associated with protein-energy wasting of skeletal muscle in diabetic kidney disease (DKD) model of Goto-Kakizaki(GK) rats and evaluate the effects of low-protein diet supplemented with α-keto acids on muscle wasting.Methods Forty-five male 24-week-age GK rats were randomly divided into three groups,normal protein diet group (NPD),low-protein diet group (LPD) and LPD +or-keto group (Keto).Fifteen gender and age matched Wistar rats were served as control group (CTL).The living condition of GK rats was observed and the weight was measured once a week.Urine albumin,serum glucose,creatinine and urea nitrogen were measured at 24,32,40,48 week age.Soleus muscle was observed to calculate the muscle size and the percentage of Ⅰ and Ⅱ type muscle fiber with software after SDH and NADH staining at 48-week-age.Tissue ultrastructure was observed under the transmission electron microscopy.The activity of citrate synthase was detected by spectrophotometer.Expression of mitochondrial DNA was examined by Q-PCR.Results Compared with the CTL group,NPD,LPD and Keto groups had lower body weight,higher urine albumin,higher serum creatinine and urea nitrogen (P ＜ 0.05).The crosssectional area of muscle fibers was larger in CTL group.Compared with CTL group,the muscle fiber was partly broken,the mitochondrial morphology was obviously changed,the percentage of type Ⅱmuscle fiber was increased significantly (P ＜ 0.05),and the activity of citrate synthase and the number of mitochondrial DNA were decreased significantly in NPD,LPD and Keto groups (P ＜ 0.05).In Keto group,muscle wasting was improved compared with NPD and LPD group (P ＜ 0.05),the crosssectional area of soleus muscle increased and the percentage of type Ⅱ muscle fiber decreased,levels of urine albumin,semm creatinine and urea nitrogen decreased (P ＜ 0.05).Under transmission electron microscopy,the muscle fiber of keto group was intact and mitochondiral morphology was close to that of CTL group.The activity of citrate synthase and number of mitochondiral DNA were higher as compared to CTL group (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,mitochondrion is swelling,the number of mitochondrial DNA is decreased and mitochondrial function is impaired.Low-protein diet supplemented with α-keto acids can improve mitochondrial damage and muscle wasting induced by DKD.
