Effect of Gushukang Granule on Osteoclast Exosomes in the Treatment of Postmenopausal Osteoporosis:A Mechanism Study
Objective To explore the underlying mechanism of Gushukang Granule(GSK)in treating postmenopausal osteoporosis(PMOP)via tuning osteoclast(OC)derived exosomes.Methods OC and osteoblast(OB)models were established.Additionally,control serum and GSK-containing serum were prepared for experimentation.Blank group,control group,control exosome(control serum)group and drug exosome(GSK-containing serum)group were set up.Western Blot analysis was used to detect OC-derived exosome levels,while the morphology of the exosomes was examined using transmission electron microscopy.Additionally,fluorescence labeling was used to investigate the uptake of exosomes by OBs.Intervention experiments were then conducted on the OB model.Alkaline phosphatase(ALP)expression was detected by ALP kit,and the gene expression of osteocalcin(OCN),osteopontin(OPN)and RUNt-associated transcription factor 2(RUNX2)were detected by Real-time PCR.PMOP rat model was established in animal experiments,and the rats were divided into sham operation group,model group,and drug experimental group,6 in each group.The model group was given physiological salt gavage,and the drug experimental group was given GSK gavage.After 8 weeks,all animals were euthanized,and their femoral tissues were collected.Hematoxylin and eosin(HE)and TRAP staining were used to investigate osteocyte differentiation in these tissues.Additionally,OCN,OPN,RUNX2,receptor activator of nuclear factor-κ B ligand(RANKL),and osteoprotegerin(OPG)protein expressions in the collected tissues were detected by Western Blot analysis.Results The OC,OB,and PMOP models were successfully established.Subsequently,interventions were performed on the OC models using the control serum and the GSK-containing serum.The CD9,CD63,and CD81 protein expressions were specifically examined,and it was found that OBs effectively absorbed OC-induced exosomes.Exosomes extracted from the control exosome group and dug exosome group downregulated mRNA expressions of OB differentiation factors such as ALP,OCN,OPN,and RUNX2,the inhibitory effect on OB differentiation was markedly reduced in the drug exosome group(P＜0.01).In vivo results showed that compared with the model group,the staining results revealed that the drug experimental group had fewer broken bone lacunae in the trabecular bone,less damage to the compact bone,and less bone resorption.Furthermore,compared with the model group,the drug experimental group exhibited significantly higher expression levels of OB markers such as OCN,OPN,and RUNX2 and significantly lower RANKL and OPG expression levels in the femoral tissue(P＜0.01).Conclusions The GSK-containing serum inhibited OC differentiation and effectively improved the inhibitory effect of OC-induced exosomes on OB differentiation.Furthermore,GSK was found to significantly accelerate OB differentiation while simultaneously decelerating OC differentiation in PMOP rats.
Gushukang Granuleosteoclastsosteoblastsexosomespostmenopausal osteoporosisChinese herbal compoundChinese patent medicine
国家科技期刊平台
NSTL
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