| We know that insufficient bone volume of the alveolar bone can lead to problems such as poor retention of the removable denture and difficulty in repairing the implant denture.Therefore,good alveolar bone mass is one of the basic conditions for successful restoration.In the clinic,osteomyelitis of the jaw,jaw cyst,benign and malignant tumor of the jaw,trauma and physiological bone resorption in the edentulous area may lead to insufficient alveolar bone volume,which will bring d'ifficulties to the repair process and affect the success of oral repairs in a long time.Therefore,finding ways to increase bone mass has become a research hotspot.Bone morphogenetic proteins(BMPs)are members of the transforming growth factor beta(TGF-?)superfamily,play a regulatory role in the proliferation and differentiation of many organs.In the 1965,Urist and other scholars extracted a class of protein components that contribute to osteogenesis from decalcified bone,there are a variety of bone morphogenetic proteins found with the function of promoting osteogenesis.BMP4 plays an important role in promoting bone tissue formation and regeneration.Some scholars have found that osteoporosis occurs after BMP4 signaling activation is inhibited in BMSCs.In vivo studies by using mouse ovariectomized models,revealing that BMP4 signaling is an important target of bone tissue regeneration and osteogenic treatment.Experiments have shown that BMP4 signaling promotes the transfer of osteoblasts from the growth plate to the ossification center,which is equivalent to the sustained motility of bone growth;bone repair materials with BMP4 also have significant osteogenesis in animal models of skull defects.In recent years,the research on osteogenesis of traditional Chinese medicine has received more and more attention.According to the theory of "The kidney is responsible for promoting bone and bone marrow production",the use of Chinese medicine for tonifying kidneys and strengthen the body is a theoretical basis of traditional Chinese medicine preventing osteoporosis and promoting osteogenesis.With the deepening of research on traditional Chinese medicine,more and more Chinese medicines and traditional Chinese medicine monomers with kidney-reinforcing effects have been confirmed to have osteogenesis effects.However,the traditional Chinese medicine is often mixed with the application of the formula,and its own extract is a mixture of various monomers.Therefore,there is a great significance to screen out the effective monomer components and achieve drug development for specific gene targets.First,we screened a variety of Chinese herbs monomers with the function of nourishing kidney that promote the transcription of the BMP4 promoter by using pGL2-BMP4 promoter luciferase reporter gene recombinant plasmids that have been successfully prepared by the our group previously.Subsequently,the regulation of BMP4 protein expression by these traditional Chinese medicines was examined by Western blot.Through the above experiments,we found that schisandrin C is effective in promoting the transcription of the BMP4 promoter and the expression of the BMP4 protein and the promoting effect is related to the drug concentration.Then,we designed a more precise concentration gradient experimental group and repeated the above experiment.We found that 10-8mol/L is the optimal concentration of schisandrin C on promoting BMP4 promoter transcription and protein expression.In order to further verify the promoting effect of Schisandrin C on the proliferation and differentiation of Mc3T3-E1 cells,we designed MTT assay and ALP assay.We found that in terms of promoting proliferation,there was no significant difference between the drug-administered group and the non-administered group on the first day and the second day after administration;from the third day,the OD value of the drug-administered group was significantly higher than that of the non-administered group(P<0.05),which showed that Schisandra chinensis had a pro-proliferative effect on Mc3T3-E1 cells.In terms of promoting differentiation,ALP was used as a detection index for differentiation,and we found that ALP activity in the drug-administered group was always higher than that in the non-administered group,which showed significant difference on the third day(P<0.05).It confirmed that schisandrin C had a certain differentiation promoting effect on Mc3T3-E1 cellsTo investigate whether BMP4 is a target for the proliferation and differentiation of Mc3T3-E1 cells,we performed a BMP4 gene silencing assay.We found that after silencing the BMP4 gene,the effect of schisandrin C on proliferation was significantly reduced,and the same result was obtained in the detection of ALP activity.Therefore,we have reason to speculate that up-regulation of BMP4 expression is a possible pathway for schisandrin C to promote proliferation and differentiation of Mc3T3-E1 cells.Through the above experiments,we concluded that Schisandra C can promote the transcription of BMP4 promoter and the expression of BMP4 protein,and its optimal concentration is 10-8mol/L.Schisandrin C also can promote the proliferation and differentiation of Mc3T3-E1 cells,and up-regulation of BMP4 expression is a possible pathway for Schisandrin C to promote proliferation and differentiation of Mc3T3-E1 cells.This provides a new idea for the clinical application of Chinese medicine monomer for osteogenic treatment. |
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