摘要
目的观察慢性肾脏病（CKD）患者骨骼肌萎缩现象，并初步探讨自噬在骨骼肌萎缩中的可能作用。方法对2012年本市4家医院确诊为CKD5期的22例尿毒症患者在腹膜透析置管术中行腹直肌活检。将8例诊断为子宫腺肌病并拟行经腹全子宫切除术患者及6例确诊为腹壁疝并拟行腹壁疝修补术患者设为对照组，于手术时留取少许腹直肌标本。所有标本行HE染色，观察肌纤维形态并计算肌纤维横截面积，透射电子显微镜检测骨骼肌自噬小体。采用RT—PCR法及Western印迹法分别检测自噬相关基因微管相关蛋白1轻链3B（LC3B）、Beelin．1及Bcl2-腺病毒E1B结合蛋白3（Bnip3）的mRNA及蛋白表达情况。结果CKD患者腹直肌肌纤维的横截面积显著小于对照组[（2982．20±629．42）μm^2比（3928．01±836．9）μm^2，t=-2．86，P〈0．051，LC3B、Beclin-1及Bnip3的mRNA表达量显著高于对照组（t=2．23、3．50、6．76，均P〈0．05），且LC3B（I及Ⅱ型）、Beclin-1、Bnip3的蛋白表达量亦显著高于对照组（t=3．51、3．97、2．46、2．49，均P〈0．05）。电镜检查发现CKD患者腹直肌中存在大量的自噬小体，而对照组少见。结论CKD患者存在骨骼肌萎缩及自噬活化现象，推测自噬活化可能参与了CKD患者骨骼肌萎缩的发生。
Objective To study skeletal muscle atrophy and the change of autophagy in skeletal muscle of patients with chronic kidney disease. Methods Mean muscle cross sectional area, mRNA and protein expression of autophagy markers Bcl-2-adenovirus E1B interacting protein 3 （LC3B）, Bcl-2-adenovirus E1B interacting protein 3 （Bnip3）, Beclin- 1 were measured in rectus abdominis biopsies obtained from 22 consecutive patients with stage 5 CKD scheduled for peritoneal dialysis from 4 hospitals in Shanghai. Control biopsies were obtained from another 8 healthy subjects during elective surgery for adenomyosis and 6 subjects during elective surgery for abdominal wall hernias. Rectus ahdominis muscles were obtained at the beginning of surgery. HE staining was performed and mean cross sectional area （CSA） was calculated. Electron microscopy was used to confirm the changes of autophagy, mRNA levels of LC3B, Beelin- 1, Bnip3 were evaluated by RT-PCR and protein levels of those parameters were evaluated by Western blotting. Results Compared with control group, mean CSA of muscle fibers was decreased and the transcript levels of LC3B, Beelin- 1, Bnip3 were up-regulated in CKD group. Similarly, protein levels of LC3BI, LC3B II, Beclin-1 and Bnip3 were increased in CKD group. Additionally, activation of autophagy was confirmed by the appearance of autophagosomes by electron microscopy. Conclusion Chronic kidney disease may cause skeletal muscle atrophy and lead to activation of autophagy, which may contribute to muscle atrophy.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2013年第5期333-338,共6页
Chinese Journal of Nephrology
关键词
肾功能不全
慢性
骨骼肌
自噬
蛋白质-能量消耗
Renal insufficiency, chronic
Muscle, skeletal
Autophagy
Protein- energy wasting
