Mechanism Of Jingui Shenqi Pills In Preventing And Treating PMOP By Regulating BMP/Smad Signaling Pathway

Objective: To study the mechanism of Jingui Shenqi Pills in preventing and treating postmenopausal osteoporosis,and to explore the application of traditional Chinese medicine theory of "warming and tonifying kidney yang,in order to generate marrow and strengthen bone" in postmenopausal osteoporosis.Methods:SPF female SD rats were randomly selected as the sham operation group,and only the same amount of fat near the ovary was removed.The other rats were ovariectomized to duplicate the rat model of postmenopausal osteoporosis.The operated rats were randomly divided into model group,Jinkui Shenqi Pills high,medium and low dose estradiol groups,8 rats in each group.The rats were weighed weekly to calculate the weight gain of rats in each group.After 3 months of administration,the rats were sacrificed and the uterine coefficient was calculated;bone mineral density and bone microstructure;HE staining was used to observe the pathological changes of the femur;automatic biochemical analyzer was used to detect the changes of calcium and phosphorus in the serum of rats in each group;Elisa method was used to detect the changes of ALP,OPG and IL-6 expression in the serum of rats in each group;The m RNA expression of BMP-2,Smad1 and Runx2 in rat bone tissues was detected by Q-PCR;the protein expression of BMP-2,Smad1 and Runx2 in rat bone tissues was detected by Western blot.Results: Compared with the sham group,the body weight gain of the model group was significant(P<0.01),and the uterine coefficient was significantly decreased(P<0.01),while the Jinkui Shenqi Pills and estradiol groups effectively reduced the body weight gain caused by OVX,and the uterine coefficient of the estradiol group was significantly increased(P<0.05).Micro-CT results showed that after 3 months of treatment,the bone mineral density of the model group was significantly decreased compared with the sham group(P<0.01);compared with the model group,the bone mineral density of the Jinkui Shenqi Pills group and the estradiol group was significantly increased(P<0.05).Three-dimensional imaging and HE staining of the femur showed that the size and thickness of the bone in the sham operation group were moderate,neatly arranged,and the bone connectivity was good;the femoral bone in the model group became shorter and thinner,sparse,fractured,and significantly osteoporotic;after treatment with Jinkui Shenqi Pills,the bone microstructure was significantly improved,and the bone structure was intact.The biochemical results showed that compared with the sham operation group,the serum calcium and phosphorus levels in the model group were significantly decreased(P<0.01);while the expression in the Jinkui Shenqi Pills model group was significantly increased(P<0.05),and there was no statistical difference between the Jinkui Shenqi Pills group and the estradiol group.Elisa results showed that compared with the sham operation group,the contents of ALP and IL-6 in the serum of rats in the model group were significantly increased,and the contents of OPG were significantly decreased;compared with the model group,the high and medium doses of Jinkui Shenqi Pills effectively decreased the contents of ALP and IL-6 in the serum and increased the contents of OPG.Q-PCR results showed that the expressions of BMP-2,Smad1 and Runx2 m RNA in the bone tissues of rats in the model group were significantly lower than those in the sham group(P<0.01);compared with the model group,the expressions of BMP-2,Smad1 and Runx2 m RNA in the bone tissues of rats in the high and medium dose estradiol groups of Jinkui Shenqi Pills were significantly increased(P<0.05).Westernblot results showed that the expression of BMP-2,Smad1 and Runx2 protein in the bone tissue of rats in the model group was significantly reduced compared with the sham group(P<0.01);compared with the model group,the expression of BMP-2,Smad1 and Runx2 protein in the bone tissue of rats in the Jinkui Shenqi Pills and Estradiol groups was significantly increased(P<0.05),and there was no statistically significant difference between the Jinkui Shenqi Pills group and the estradiol group.Conclusions: Jingui Shenqi Pills can increase bone mineral density in order to improve osteoporosis.The mechanism may be through regulating BMP/Smad signaling pathway and enhancing the expression of osteogenic factors BMP-2,Smad1 and Runx2.