补肾健脾活血方基于自噬-凋亡双通路改善去势大鼠骨质疏松和肌肉萎缩的机制研究OA
Mechanistic Study of Bushen Jianpi Huoxue Formula in Improving Osteoporosis and Muscle Atrophy in Castrated Rats via the Autophagy-Apoptosis Dual Pathway
目的:探讨补肾健脾活血方通过自噬-凋亡双通路改善去势大鼠骨质疏松和肌肉萎缩的机制.方法:60只雌性SD大鼠随机分为正常组(Control组)、假手术组(Sham组)、模型组(OVX组)、补肾健脾活血方组(OVX+BSF组,2.979 g/kg)和雌二醇组(OVX+E2 组,0.104 mg/kg).OVX组、OVX+BSF组和OVX+E2组采用双侧卵巢切除法造模,造模成功后,给予相应药物干预.每周按时测量大鼠体质量和前爪抓力;灌胃3个月后,采用骨密度仪检测大鼠体成分、全身和局部骨密度;micro CT检测大鼠股骨远端松质骨形态变化;HE染色观察股骨和腓肠肌病理改变;免疫荧光检测股骨LC3B的荧光强度;透射电镜检测股骨自噬小体数量变化;TUNEL荧光染色检测股骨凋亡水平;qRT-PCR检测股骨凋亡基因表达水平.结果:与Control组和Sham组比较,OVX组大鼠前爪抓力、骨骼肌质量指数、全身及局部骨密度明显下降,股骨远端松质骨结构破坏严重;与OVX组比较,OVX+BSF组去势大鼠的前爪抓力、骨骼肌质量指数、全身及局部BMD升高,股骨远端骨微结构改善,股骨自噬蛋白LC3B表达降低,自噬小体数量减少,凋亡细胞减少,抗凋亡基因Bcl-2和Bcl-xL表达增加,促凋亡基因Bad表达降低.结论:去势可引起大鼠骨质疏松和肌萎缩,BSF明显改善去势大鼠的骨质疏松和肌萎缩,作用机制可能与降低大鼠股骨自噬-凋亡水平有关.
Objective:To explore the mechanism by which Bushen Jianpi Huoxue Formula improves osteoporosis and muscle atrophy in castrated rats through the autophagy-apoptosis dual pathway.Methods:Sixty female SD rats were randomly divided into five groups:Control group,Sham group,OVX group(model group),OVX+BSF group(2.979 g/kg),and OVX+E2 group(0.104 mg/kg).OVX,OVX+BSF,and OVX+E2 groups were subjected to bilateral ovariectomy to establish the model.After successful modeling,the corresponding drug interventions were administered.Rat body weight and forepaw grip strength were measured weekly.After 3 months of gavage,bone composition,whole-body and local bone mineral density(BMD)were assessed using a bone densitometer;micro-CT was used to detect changes in trabecular bone morphology at the distal femur;HE staining was performed to observe pathological changes in the femur and gastrocnemius muscle;immunofluorescence was used to measure LC3B fluorescence intensity in the femur;transmission electron microscopy was used to detect changes in the number of autophagosomes in the femur;TUNEL fluorescence staining was used to assess apoptosis levels in the femur;and qRT-PCR was used to determine the expression levels of apoptosis-related genes in the femur.Results:Compared to the Control and Sham groups,the OVX group exhibited significantly decreased forepaw grip strength,muscle mass index,and whole-body and local BMD,with severe damage to the trabecular bone structure at the distal femur.Compared to the OVX group,the OVX+BSF group showed increased forepaw grip strength,muscle mass index,and whole-body and local BMD,with improved bone microstructure at the distal femur.The expression of the autophagy protein LC3B and the number of autophagosomes were reduced,while apoptotic cells decreased.The expression of anti-apoptotic genes Bcl-2 and Bcl-xL increased,while the pro-apoptotic gene Bad decreased.Conclusion:Castration induces osteoporosis and muscle atrophy in rats.Bushen Jianpi Huoxue Formula significantly improves osteoporosis and muscle atrophy in castrated rats,likely through its effects on reducing autophagy-apoptosis levels in the femur.
马江涛;涂潇雅;叶茂林;柴爽;刘源;黄宏兴;王吉利;李小玉
河南省洛阳正骨医院(河南省骨科医院),河南 郑州 450046郑州人民医院,河南 郑州 450003广州中医药大学第三附属医院,广东 广州 510375河南省中医院,河南 郑州 50053
补肾健脾活血方去势大鼠骨质疏松肌肉萎缩自噬凋亡
Bushen Jianpi Huoxue FormulaCastrated ratsOsteoporosisMuscle atrophyAutophagyApoptosis
《中医药信息》 2024 (010)
19-26 / 8
河南省自然科学基金青年科学基金项目(222300420198);河南省重点研发与推广专项(科技攻关)(242102310522,242102310539)
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