Mechanistic Study of Bushen Jianpi Huoxue Formula in Improving Osteoporosis and Muscle Atrophy in Castrated Rats via the Autophagy-Apoptosis Dual Pathway
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.