Bushen Huoxue Fufang Tenghuang Jiangu Capsule Attenuates Postmenopausal Osteoporosis via Activating Sirt1/FoxO3a in Ovariectomized Rats

OBJECTIVE To observe the effects of Bushen Huoxue Fufang Tenghuang Jiangu Capsule on the expression levels of Sirt1, FoxO3a, and the expression levels of autophagy molecules P62, Beclin1, and LC3-Ⅱ, so as to investigate the mechanism of Bushen Huoxue Fufang Tenghuang Jiangu Capsule in postmenopausal osteoporosis rats.

METHODS  The rats postmenopausal osteoporosis model was established using bilateral ovariectomy method, the general situation was observed. The microstructure changes were observed, and the bone volume fraction(BV/TV) , the trabecular thickness(Tb.Th) of femoral bone was checked using micro CT. The ALP, CTX-Ⅰ and BGP content were detected by ELISA method. The gene levels of Sirt1, FoxO3a, and P62 were detected using real time fluorescence quantitative PCR. The expression of Sirt1, FoxO3a, P62, Beclin1, LC3-Ⅱ was analyzed using Western blotting.

RESULTS  The general situation and weight results showed that the general situation significantly improved, the weight significantly reduced in the high-dose group of Bushen Huoxue Fufang Tenghuang Jiangu Capsule(P<0.05). The micro CT results showed that the Bushen Huoxue Fufang Tenghuang Jiangu Capsule could significantly reduce the degree of femoral bone microstructure damage, the medium-dose and high-dose groups of the Bushen Huoxue Fufang Tenghuang Jiangu Capsule could significantly increase BV/TV(P<0.05 or P<0.01), while the high-dose group could significantly increase Tb.Th(P<0.01). The detection results of dual energy X-ray absorptiometry showed that the Bushen Huoxue Fufang Tenghuang Jiangu Capsule could significantly increase bone mineral density(P<0.01). The ELISA results showed that the ALP contents significantly increased in the each-dose group of Bushen Huoxue Fufang Tenghuang Jiangu Capsule(P<0.01), the BGP contents significantly increased in the the medium-dose and high-dose groups of Bushen Huoxue Fufang Tenghuang Jiangu Capsule(P<0.01), the CTX-Ⅰ contents significantly decreased in the high-dose group of Bushen Huoxue Fufang Tenghuang Jiangu Capsule(P<0.01). The detection results of gene and protein of key molecules in signal axis showed that the gene and protein expressions of key molecule Sirt1, FoxO3a in the Sirt1/FoxO3a signal axis significantly raised in the each-dose group of Bushen Huoxue Fufang Tenghuang Jiangu Capsule(P<0.05 or P<0.01). The detection results of gene and protein of autophagy molecule showed that each-dose group of Bushen Huoxue Fufang Tenghuang Jiangu Capsule could significantly reduce the gene and protein expression of autophagy molecule P62(P<0.01), and significantly increase the protein expression of autophagy molecule Beclin1(P<0.05 or P<0.01), the medium-dose and high-dose groups could significantly increase the protein expression of autophagy molecule LC3-Ⅱ(P<0.01).

CONCLUSION Bushen Huoxue Fufang Tenghuang Jiangu Capsule can alleviate microstructural damage to femoral bones and treat postmenopausal osteoporosis, possibly through activating the Sirt1 and FoxO3a expression of key molecules in Sirt1/FoxO3a signal axis, and further down-regulating the expression level of autophagy molecule P62 and upregulating the expression levels of autophagy molecules Beclin1 and LC3-Ⅱ.