| Objective:Based on the establishment of an animal model of postmenopausal osteoporosis,the mechanism of daphnetin in the treatment of postmenopausal osteoporosis was explored based on the inflammatory related factor regulation pathway TRAF6/NF-κBp65/NFATc1,and the factors related to inflammation and bone metabolism were used as the observation indexes,so as to provide experimental basis for the prevention and treatment of osteoporosis.Methods: Sixty 3-month-old female SD rats were randomly divided into: daphnetin low(group A),daphnetin medium(group B),daphnetin high dose group(group C),control group(group D),model group(group E)and blank group(group K),with 10 rats in each group.Ovariectomy was performed in all rats except the blank group.Building after the success of low,medium and high dose groups,respectively be daphnetin 35mg/kg·d,70mg/kg·d,140mg/kg·d,the control shall be Allen phosphonic acid sodium lavage 7.3mg/kg· week,model group normal saline70 mg/kg·d to fill the stomach,not to interfere in blank group,continuous lavage after 12 weeks,bone mineral density using dual energy X-ray detection rats,rats using Elisa to detect serum Ca,P,ALP,IL-6,TNF alpha,PICP,CTX1,the expression of PTH.The protein expressions of TRAF6,NF-κBp65 and NFATC1 were detected by WB,and the gene expressions of TRAF6,NF-κBp65 and NFATC1 were analyzed by RT-PCR Results:1.Dual energy X results showed that after 12 weeks of administration,the BMD of medium dose,high dose,control group and blank group were all higher than that of model group,and the difference was statistically significant(P<0.05).2.ELISA results showed that IL-6 content in low-dose and high-dose groups was lower than that in model group and higher than that in blank group and control group,and the differences were statistically significant(P<0.05);The content of TNF-α in low dose,high dose,control group and blank group was lower than that in model group,and the difference was statistically significant(P<0.05).The content of Ca in high-dose group was higher than that in control group and model group,and the difference was statistically significant(P <0.05;P content in low-dose,medium-dose and high-dose groups was higher than that in model group and blank group,and lower than that in control group,and the difference was statistically significant(P < 0.05).ALP content in medium-dose and high-dose groups was lower than that in model group,and the difference was statistically significant(P < 0.05).PICP content in low-dose,medium-dose and high-dose groups was higher than that in model group,and the difference was statistically significant(P < 0.05).The difference of CTX1 content between low-dose group and control group,model group and blank group was statistically significant(P < 0.05).The difference of CTX1 content between medium-dose group and control group and blank group was statistically significant(P <0.05).Compared with model group,the content of PTH in low-dose and high-dose groups had statistical significance(P < 0.05).4.Western blot results showed that the protein content of TRAF6 in low-dose and medium-dose groups was lower than that in control group,model group and blank group,and the difference was statistically significant(P < 0.05).The protein content of TRAF6 in high-dose group was significantly different from that in model group(P < 0.05).The protein content of NF-κBp65 in low-dose and medium-dose groups was lower than that in control group,model group and blank group,and the difference was statistically significant(P < 0.05).The protein content of NF-κBp65 in high-dose group was lower than that in model group,and the difference was statistically significant compared with that in control group and model group(P < 0.05).Compared with the control group,model group and blank group,the protein content of NFATC1 in low-dose and medium-dose groups had statistical significance(P < 0.05).4.RT-PCR results showed that the expression levels of TRAF6,NF-κBp65 and NFATC1 in low-dose,medium-dose and high-dose groups were significantly different from those in model group and blank group(P < 0.05).Conclusion:1.Daphnetin can reduce the expression of IL-6 and TNF-α inflammatory factor in serum of osteoporosis rats.2.Daphnetin can improve bone metabolism and alleviate the process of osteoporosis by inhibiting the bone resorbing factor CTX1、 ALP and increasing the bone forming factor PICP in osteoporosis rats3.Daphnetin can inhibit the inflammatory response and the activation of osteoclasts in rats by inhibiting the expression of TRAF6 /NF-κBp65/ NFATC1 pathway protein and gene,thus inhibiting bone resorption,improving bone metabolism and playing an antiosteoporosis role.These results suggest that daphnetin may be a potential new drug for the treatment of postmenopausal osteoporosis. |
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