去卵巢大鼠肌肉衰减症模型构建及雌激素补充治疗的实验研究  

Establishment of rat model of sarcopenia by ovariectomy and estrogen replacement therapy

作　　者：黄昱彬 凌丽[1] 熊正爱[1] HUANG Yubin;LING Li;XIONG Zheng'ai(Department of Gynecology and Obstetrics,the Second Affiliated Hospital of Chongqing Medical University,Chongqing,400010,China)

机构地区：[1]重庆医科大学附属第二医院妇产科,重庆400010

出　　处：《陆军军医大学学报》2023年第18期1937-1946,共10页Journal of Army Medical University

摘　　要：目的建立SD大鼠绝经后肌肉衰减症模型,观察雌激素补充对肌肉衰减症的治疗作用。方法①44只12周龄雌性SD大鼠(体质量220±20 g)按体质量分层随机抽样分为假手术(SHAM)组、去卵巢(ovariectomized,OVX)组,每组22只。OVX组行双侧卵巢切除术,SHAM组行假手术,监测体质量,电子抓力仪测量大鼠前肢抓力,12周后通过双能X射线分析仪(DXA)分析身体成分,通过相对瘦质量判定建模是否成功,而后每组各处死10只取腓肠肌,用腓肠肌质量/体质量得出肌肉衰减症指数(sarcopenia index,SI),HE染色观察腓肠肌肌纤维横截面积,电镜下观察肌原纤维超微结构。②余下大鼠进一步分为假手术(SHAM)组、假手术+油剂注射(SHAM+O)组、去卵巢(OVX)组、去卵巢+雌激素补充(OVX+E_(2))组,每组6只,OVX+E_(2)组每日予以17β-雌激素腹部皮下注射,SHAM+O组每日予以10%DMSO+90%芝麻油皮下注射,监测体质量、前肢抓力,干预8周后DXA分析体成分,ELISA法测量血清E_(2)、FSH,HE染色观察腓肠肌肌纤维横截面积,电镜下观察肌原纤维超微结构,免疫荧光染色分析Bax、Bcl-2的表达。结果①建模12周后,OVX组相对瘦质量显著降低(P<0.001),提示建模成功,体质量(P<0.001)、体脂率显著升高(P<0.001),相对抓力(P<0.001)、SI值、肌纤维横截面积显著降低(P<0.01),电镜下线粒体密度降低(P=0.012)。②经雌激素补充治疗后,OVX+E_(2)组抓力、相对瘦质量、体质量、体脂率、肌纤维横截面积、电镜下线粒体密度均恢复到SHAM组水平;免疫荧光显示OVX组较SHAM与OVX+E_(2)组Bcl-2/Bax显著降低(P<0.01)。结论成功建立了大鼠绝经后肌肉衰减症模型,雌激素补充治疗能显著改善去卵巢大鼠的肌肉衰减,可能与雌激素抑制肌细胞凋亡有关。Objective To establish a rat model of postmenopausal sarcopenia and explore the therapeutic effect of estrogen supplementary therapy on the disease.Methods①Forty-four 12-week-old female SD rats(220±20 g)were randomly divided into sham group and ovariectomized(OVX)group by stratified random sampling based on body weight,with 22 rats in each group.The rats of the OVX group were given bilateral ovariectomy,while those of the SHAM group underwent sham surgery.The body weight was monitored,and the grip strength of the forelimb was measured using an electronic grip strength meter.In 12 weeks after surgery,the body composition was analyzed by X-ray absorptiometry(DXA)to identify the model establishment by relative lean mass.Ten rats in each group were sacrificed to harvest the gastrocnemius,and then sarcopenia index(SI)was calculated as the ratio of gastrocnemius mass to body weight.HE staining and transmission electron microscopy were applied respectively to observe the cross-sectional area of the muscle fibers and the structure of myofibrils were determined.②The remaining rats were further divided into SHAM group,SHAM+O group,OVX group and OVX+E_(2)group,with 6 rats in each group.17β-estradiol was injected subcutaneously in OVX+E_(2)group,while control solvent(10%DMSO+90%sesame oil)was injected subcutaneously in SHAM+O group.The grip strength and body composition were measured and body composition was analyzed.ELISA was used to measure the serum levels of E_(2)and FSH,HE staining was employed to observe the cross-sectional area of gastrocnemius muscle fibers,electron microscopy was conducted to observe the ultrastructure of myofibrils,and immunofluorescence staining was applied to analyze the expression of Bax and Bcl-2.Results①After 12 weeks of modeling,the rats in the OVX group had significant lower relative lean mass(P<0.001),indicating the successful establishment of sarcopenia model,and had obviously increased body weight(P<0.001)and body fat rate(P<0.001),but decreased relative grip(P<0.001),SI value,ave

关 键 词：肌肉衰减症 雌激素替代疗法 大鼠模型 卵巢切除术 凋亡 

分 类 号：R589.305[医药卫生—内分泌] R711.51[医药卫生—内科学] R977.12[医药卫生—临床医学]