| Osteoporosis is one of common bone disorders,affecting millions of people worldwide.Treatments of osteoporosis consist of pharmacotherapy and non-pharmacological interventions,such as mineral supplementation,lifestyle changes,and exercise programs.Due to the minimum side effects and favorable cost-effective therapeutic effects,herbal medicine has been widely applied in clinical practices for more than 2,000 years in China.Rhizoma drynariae is used commonly in the treatment of osteoporosis and bone nonunion in traditional Chinese medicine.Modern pharmacological research indicates that naringin is the main effective component of rhizoma drynariae,which can accelerate osteoblast proliferation and differentiation,and inhibiting osteoclast formation,and can also suppress the OP induced by drugs.So we speculated the naringin which has a dual role can correct the OP state of imbalance between osteoblasts and osteoclasts,which has the effect of the OP resistance.In this experiment,we will be taking the research of the above,the main part of the points above are as follows:1.Effects of naringin on the proliferation and osteogenic differentiation of rat calvarial osteoblasts.(1)Objective: To study the effects of naringin on the proliferation and osteogenic differentiation of mouse calvarial osteoblasts.(2)Methods: Four-day-old SD mouses(Experimental Animal Center of Chongqing Medical University)were used in the present study.Primary cultured calvarial osteoblasts from newborn mice are used as the model of late stage osteoblasts.The medium was changed every 3 days.After reaching 80%confluence,the cells were cultured in normal medium which mainly contains ?-MEM and 10% FBS(control)and various concentrations of naringin(0,1,10 mg/L).The medium was changed every three days.At 24,72,243 h post treatment with naringin,CCK-8 assay was performed.At 24 h alizarin red staining was performed.Treated with naringin for1,3,9days,cells were measured with alkaline phosphatase activity.OCN,OPN protein level was detected between the naringin group and control group using Elisa method.Total RNA was isolated from ROB using TRIZOL reagent.The RNA samples extracted were analyzed for markers of osteogenic differentiation: ALP,osteopontin,and osteocalcin.(3)Results: The result of ALP activity showed that dose-dependent naringin increased the ALP activity.The cells in naringin solution presented higher activity,and the 1 mg/L group showed the most significant differences.We also found that naringin can increase two osteogenic protein level(OC,OPN).The RT-PCR result for the detection of osteoblastic differentiation markers showed that all osteogenic differentiation genes expression were increased in naringin solutions as compared to the control group.The dose-dependent naringin(0,1,10 mg/L)increased the expression of these osteogenic differentiation markers(ALP,osteopontin,osteocalcin).The result of Alizarin red staining indicated that there was a greater calcium node formation ratio in the naringin groups than in the control one.(4)Conclusion:Naringin can enhance the proliferation and osteogenic differentiation of ROB.It is interesting to note that in our experiment,a concentration that at 1mg/L will have a significant influence.So we speculated the naringin has the effect of the OP resistance.2.Effects of Naringin on the Formation and Function of Osteoblasts and Osteoclasts in mouse calvarial organ culture.(1)Objective: Preliminary study on the mechanism of the balance between coexisting bone cells by using bone organ culture with naringin.(2)Methods:Four-day-old mouses(Experimental Animal Center of Chongqing Medical University)were used in the present study.Calvarial organ from newborn mice are used as the model,cultured in ?-MEM.late stage osteoblasts.Various concentrations of naringin(0,1,10 mg/L)were treated..Using HE staining,TRAP staining,calcium ion concentration detection,PCR detection of OPG,RANKL m RNA levels(3)Results: The effect of naringin on the formation of new bone was significantly increased after HE staining,and the thickness of slice bone was significantly increased.The increase of 1 mg / L was the most significant,followed by 10 mg / L concentration.With the passage of time,the effect of naringin on bone organ.Trap staining is more and more shallow,indicating that the number of osteoclast coloring has decreased.RT-PCR results showed that the expression of osteogenic differentiation gene OPG was increased compared with the control group,and the expression of RANKL was decreased.(4)Conclusion:The study of bone organ culture model showed that naringin had the effect of promoting osteoblasts,and also had the effect of inhibiting osteoclasts.The mechanism was related to RANKL / OPG / RANK.3 Combines micro CT and bone organ culture techniques for the study of Chinese medicine(1)Objective : Microscopic analysis of bone formation and bone resorption(2)Methods:Four-day-old SD mouses(Experimental Animal Center of Chongqing Medical University)were used in the present study.Calvarial organ from newborn mice are used as the model,cultured in ?-MEM.late stage osteoblasts.Various concentrations of naringin(0,1,10 mg/L)were treated.Using micro CT,take an analysis of bone formation and bone resorption.(3)Results: There were almost no significant changes in the BS under different concentrations of naringin at different time points.There was no significant difference between the 1 mg / L concentration group and the control group only on the 16 th day and the 23 th day Statistically significant(P <0.05).Compared with the control group,the BV and BS / BV of the different concentrations of naringin were significantly changed at different time points,and the difference was statistically significant(P <0.05)Day,23 days,all groups showed a time effect,there was statistical significance,that is,bone volume increased significantly.1 mg / L concentration group and the control group the difference was statistically significant(P <0.01).The difference between the 10 mg /L group and the control group was statistically significant(P <0.05).(4)Conclusion:Using the neonatal mouse cranial organ as a research model,the model was successfully modeled..In the three-dimensional reconstructed images,we found that the cranial gland at the same time appeared osteogenesis and osteoclast effect(ie,bone resorption),indicating that bone organ culture is conducive to retain a variety of cells(osteoblasts,osteoclasts and bone cells)between Interrelated,and also beneficial to maintain the natural relationship between cells and extracellular matrix..Naringin treated,the thickness of the cranial bone has increased. |
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