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AIM: The effects of YIGU capsule on proliferation and IGF-I mRNA protein expressions in osteoblasts were studied. METHODS: (1) Forty 12-month old Sprague-Dawley female rats were divided randomly into four groups (YIGU capsule high dose group, medium dose group and low dose group; saline group), the drug-containing serum and control serum were prepared. (2) The new-born Sprague-Dawley rat osteoblasts were cultured with different YIGU capsule drug-containing serum at different concentrations and different exposure time. MTT method was used to observe proliferation of osteoblasts. (3) RT-PCR method was used to measure the relative IGF-I mRNA levels and ELISA method was used to measure IGF-I secretion at different exposure time. (4) ELISA method was used to measure IGF-I secretion at different exposure time. RESULTS: (1) Proliferation of osteoblasts was more than the control groups after 48, 72 and 96 h, respectively (P
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AIM: The effects of Yigu capsule on tartrate resistant acid phosphatase (TRAP) secretion and apoptosis in rat osteoclasts were investigated in order to further explore its mechanism of preventing and treating osteoporosis. METHODS: (1) Twenty-month-old Sprague-daweley rats were randomly divided into two groups(Yigu capsule group and saline group), and the drug-containing serum and control serum were prepared. (2) The newborn Sprague-daweley rat osteoclasts were cultured with different concentrations of Yigu capsule drug-containing serum. At different time point, TRAP activity was measured and the survival osteoclast was counted under reverse microscope.The percentage of osteoclast apoptosis was observed under fluorescence microscope after acridine orange staining. RESULTS: TRAP activity was lower and the percentage of osteoclast apoptosis was higher in drug-containing serum group than in control group at 24, 48 and 72 h( P
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AIM:To observe the effect of TNF-? and Yigu capsule drug-containing sera on osteoblasts apoptosis in osteoblasts-osteoclasts coculture system.METHODS:(1)Twenty female Sprague-Dawley rats,10 months old,were randomly assigned into 2 groups,NS and Yigu capsule groups,to prepare blank sera and drug-containing sera.(2)The DNA gel electrophoresis method was used to detect apoptosis of osteoblasts treated with different concentration of TNF-? in order to determine the best dosage in the co-culture system.(3)The cells were divided into four groups,TNF-? group,normal group,TNF-? + blank serum group and TNF-? + drug-containing serum group.DNA gel electrophoresis,acridine orange staining and flow cytometry were used to observe osteoblast apoptosis in these groups.RESULTS:(1)After induced by TNF-? for 72 h at a concentration of 60 ?g/L,relatively typical DNA ladder appeared in TNF-? group.(2)Only two DNA brands appeared and most of cells were well-proportioned stained in TNF-? + drug-containing serum group,the rate of osteoblasts apoptosis in TNF-? + drug-containing serum group(9.60%?0.26%)was obviously lower than that in the TNF-? group(26.90%?0.06%)and TNF-? + blank serum group(18.10%?0.06%).CONCLUSION:TNF-? induces osteoblast apoptosis in the co-culture system,and Yigu capsule drug-containing serum prevents osteoblast apoptosis induced by TNF-?.
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AIM: To investigate the effect of yigu capsule drug-containing serum on proliferation, ALP activity and expression of interleukin-11 mRNA in Sprague-Dawley rat osteobalsts in the co-culture system. METHODS: (1) Osteoblasts and osteoclasts were isolated from 1 and 5-day-old Sprague-Dawley rats, respectively. The osteoblasts-osteoclasts co-culture system was built to prevent the two kinds of cells from contact and allow the media to exchange. The experiment included two groups, drug-containing sera group and control group. (2) The osteoblasts proliferation, ALP activity and expression of interleukin-11 mRNA were detected by the MTT, 4-aminoantipyrine spectrometric methods and FQ-PCR,respectively. RESULTS: In drug-containing sera group, the osteoblasts proliferation, ALP activity, expression of IL-11 mRNA were higher than those of control group (P
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AIM:To analyze the regulatory effect of Yigu capsule on core binding factor alpha 1 (cbf ?-1) gene expression in bone of ovariectomized osteoporosis (OP) rats. METHODS: Thirty-six 10-month old Sprague-Dawley female rats were randomized into three groups: sham-operated group, model group and drug group. After intervention by the corresponding methods, the femurs were collected, SYBR green Ⅰ fluorescence quantitative PCR technique was applied with the internal control of GAPDH according to the relative quantitative formula (2-Ct), the differentially expressed multiples between the model group or drug group and sham-operated group were calculated. RESULTS: Quantitative formula analysis showed that the level of cbf ?-1 gene expression in bone of model groups was decreased than that in sham-operated rats [compared with sham-operated group, P0.05). CONCLUSION: The results of this study show that cbf ?-1 gene expression in bone tissue of ovariectomized OP rat is decreased, and Yigu capsule increases the level of cbf ?-1 gene expression in bone of OP, indicating that Yigu capsule induces bone marrow mesenchymal stem cells into osteoblasts.