| Objective1. To explore the regularties of traditional Chinese medicine treatment of postmenopausal osteoporosis.2. To explore the mechanism of Curculigoside promoting bone formation.Methods1. The clinical literature of traditional Chinese medicine therapy for postmenopausal osteoporosis which met the inclusion and exclusion criteria were collected to establish a database with EXCEL2010 software. Each article is a record. we input the source of literature, fist author and list each Chinese herb medicine from the Chinese herbal compounds, and recorded the dosage of the Chinese herb. Gram was used as the dosage unit. The Chinese traditional medicine in our database were being regularized according to the teaching material "Traditional Chinese pharmacology" and "Chinese traditional medicine name" which published in 2010. The screening prescriptions were input to the table of ECXEL 2010 to create a new database of traditional Chinese medicine. Using frequency analysis method to analyse the data, and calculate the frequency of each medicine. Every Chinese herb was be classified to the suitable category. The drugs which were used more than 10 times or more were being analysed by cluster analysi according to the herbal efficiency.2. The UMR106 cells were cultured in the medium containing penicillin-streptomycin (10000U/ml) and 10%fetal bovine serum, then put the cells into the culture incubator in 37 C and 5%CO2 to continue culturing. The medium was changed about 2-3 days according to the growth of cells. The cells in iogarithmic growth phase were seeded in 96 hole plate. The cells’ proliferation was colorimetrically assed by MTT. Cellular morphology was observed under the microscope. Alizarin red staining was performed to identify mineralization of cells. The cells in iogarithmic growth phase were cultured were used in experiment. They were divided into seven groups, namely blank control group, estrogen group, high dose of Curculigoside group, low dose of Curculigoside group, estrogen-estrogen receptor inhibitor group, high dose of Curculigoside-estrogen receptor inhibitor group, low dose of Curculigoside-estrogen receptor inhibitor group. The different groups were intervention by corresponding drugs, the cells proliferation were records by MTT. After drug intervention 24h,48h,72h, the ALP activity of cells were detected, respectively. The concentration of BGP was detected after drug intervention the 4th and the 8th day. The logarithmic phase cells were cultured and divided into four groups, namely blank control group, estrogen group, high dose of Curculigoside group, low dose of Curculigoside group. The protein expression of ER a and ER β of UMR106 cells were determined by Western blot.Rasults1. Research on Data MiningThere were 13 categories (total 138 herbs and 1773 counts of frequency) in the compound herbal treating PMOP. The herbs which were used for more than 10 times were total 8 categories, including 41 herbs. The counts of frequency were 1521. The commonly used drugs for treating PMOP were traditional Chinese Medicine "Reinforcing agents". Kidney drugs were the mainly first-line drugs, and in kidney yang drugs. In addition, the first-line drugs included Rehmannia, angelica, Salvia miltiorrhiza ect. In addition to Reinforcing agents, the second-line drugs also included tonifying drugs, and Qufengshi, strong bones, reinforcing Kidney and incigorating Sleep, clearing heat and activating blood and soothe the nerves, relieving effect.2. The basic experimental research①The results of proliferation at 24h＞48h、72h:Being compared to the control group, Estrogen group, low dose of curculigoside group and high dose of curculigoside group all can promote the proliferation of UMR106 cells (P＜0.01), but the Estrogen+inhibitor group＞high dose of curculigoside+ inhibitor group、low dose of curculigoside+inhibitor group had no difference (P＞0.05). The effect of high dose of curculigoside was more greater than the estrogen group and low dose group of Curculigoside (P＜0.01). After adding the estrogen receptor inhibitor can inhibit the proliferation of UMR106 cells (P＜0.01). ②The results of ALP activity at 24h、48h、72h:The ALP activity in estrogen, low dose of curculigoside and high dose of curculigoside group was higher than it in the control group (P＜0.01), but the Estrogen+inhibitor group、high dose of curculigoside+inhibitor group、low dose of curculigoside +inhibitor group had no difference compared with the control group (P＞0.05).And the ALP activity in the high dose of curculigoside group was higher than it in the estrogen group (P＜0.05). After adding the estrogen receptor inhibitor, ALP activity can be significantly inhibit than the group without adding the estrogen receptor inhibitor (P＜0.01). ③The results of the concentration at 4d、8d:The concentration of BGP in estrogen, low dose of curculigoside and high dose of curculigoside group was higher than it in the blank control group (P＜0.01).And The concentration of BGP in the high dose of curculigoside group was higher than it in the estrogen group (P＜0.01). ④The protein expression of ERa in the high dose of curculigoside was higher than the blank control group (P<0.05), but there was no significant difference among the estrogen group, low dose of curculigoside group and the blank control group. The protein expression of ER β was no difference among the four groups.Conclusion1. The results of the descriptive analysis were consistent with the cluster analysis results. The commonly used drugs for treating PMOP were traditional Chinese Medicine "Reinforcing agents", anlong which the recuperating kidney yang drugs was the most common. It suggest that warming the kidney to strengthen yang is the most commonly treatment principle that was used to treat the postmenopausal osteoporosis. The results indicated that kidney yang deficiency was the most mainly syndrome in postmenopausal osteoporosis. In addition, both Replenishing Kidney and Essence and activating blood circulation to dissipate blood stasis were the commonly treatment principle. It indicated that the syndrome also included kidney essence deficiency and clood stasis.2. Xianmao was the first-line drug, and its function was invigorating kidney and strengthen yang3. Curculigoside may be through the role of estrogen receptor and regulate the proliferation and differentiation of osteoblasts. Curculigoside may interact with estrogen receptor ER a to regulate the proliferation and differentiation of osteoblasts. |
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