| Background:Clinically,bone injury is very common,it is a clinical frequently occurring and common disease,which brings great suffering to patients.Fracture is the most common bone injury.Fracture nonunion or delayed union brings great pressure to patients,and consumes a lot of manpower,material and finacial resources.With age comes irreversible osteoporosis,loss of bone mass and changes in bone microstructure,and a significant increase in the risk of fracture.The key factors of fracture are the stable fixation of the fracture end and the good local blood supply to promote the repair of the damaged part.Osteoporosis can occur at any age,but it is more common in older people,especially in older women after menopause.Osteoporosis has become a worldwide problem.This disease is one of the diseases that cause serious harm to human health after cancer and cardiovascular diseases.With the increasing number of elderly people and brings inconvenience to their daily life.With the increase of bone fragility of elderly people,fractures are more likely to occur.Osteoporotic fracture belongs to a kind of pathological fracture.Due to the existence of basic diseases,the increase of bone fraglity,though a slight external force,it can cause fracture,which seriously affects the health and quality of life ofthe elderly.Once the elderly have secondary fractures,they need to stay in bed,which is prone to various complications,such as lung infections and bedsores.Because of the lack of local blood supply,the fracture is difficult to heal,which may lead to disability.Therfore,the prevention and treatment of osteoporotic fracture is particularly important.Salvianolic acid B(SalB)is the root and rhizome extract of Salvia miltiorrhiza,which is condensed by 1 molecule of caffeic acid and 3 molecules of Danshensu.Salvia miltiorrhiza has the effect of promoting blood circulation,regulating meridians,removing stasis,relieving pain,cooling blood and eliminating carbuncle.It is often used in the treatment of chest pain,abdominal pain,heat accumulation,insomnia,irregular menstruation,dysmenorrhea,amenorrhea,swelling and pain.The treatment of osteoporotic fracture by TCM syndrome differentiation is divided into three stages.In the early stage,blood stasis and swelling should be eliminated,blood stasis and swelling should be eliminated,so that blood stasis can be removed,new blood can be generated.In addition,there is a research have also shown that salB can promote the osteogeic differentiation of human bone marrow mesenchy stem cells(hBMSCs)through the activation of ERK signaling pathway.MSCs are widely used in a variety of tissue and organ repair engineering due to their strong amplification abilility under the condition of i vitro culture,and they are one of the very ideal seed cells.With stable phenotype,mesenchymal stem cells can repair bone damage with high quality after implantation in vivo.At the same time,mesenchymal stem cells have the potential of osteogenic differentiation and can also be used as gene carriers,making them a promising source of seed cells for bone tissue repair engineering.Osteogenic differentiation and chondroblast differentiation are inseparable processes,which interact with each other to complete fracture repair.ObjectiveTo observe the internal mechanism of the effect of salvianolic acid B,one of the active monomers of Salvia miltiorrhiza,on the osteogenic differentiation of human bone marrow mesenchymal stem cells(hBMSCs),so as to promote angiogenesis and enhance the osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs),so as to provide theoretical and experimental basis for the treatment of osteoporotic fracture by activating blood circulation and removing blood stasis.Methods1.The primary human bone marrow mesenchymal stem cells were cultured in α-MEM complete culture medium,and the cells of the 3-8 generation(p3-8)were transferred for experiment.The cell growth was observed under an inverted microscope,and the cell growth process was captured and recorded.2.Total RNA was collected from the cells after drug intervention and the control group,and real-time fluorescence quantitative PCR was performed to screen out the target genes and signal molecules with different expressions.3.The small interfering RNA(siRNA)interference technology was used to reduce the expression of target genes.After the siRNA interfering target genes were detected by real-time quantitative PCR,the expression levels of osteogenic genes such as ALP,BMP2 and OCN in this group of cells were detected.4.Immunofluorescence was used to detect the expression of Endomucin(EMCN)HIF1-alpha(HIF1A)and other proteins in adult bone marrow mesenchymal stem cells 48 hours after treatment with salB.5.A rat model of osteoporotic fracture was established,and salvianolic acid B aqueous solution was injected locally into the fracture to further investigate the therapeutic effect of salB in vivo.Results1.Human bone marrow mesenchymal stem cells cultured in vitro had active proliferation and good growth status.2.Real-time quantitative PCR results showed that the expression of long non-coding RNA-MALAT1(IncRNA-MALAT1)was differentially expressed in the drug intervention group and showed an upward trend.3.Real-time quantitative PCR results showed that interference with lncRNA-MALAT1 gene expression could inhibit osteogenic differentiation of human BMSCs.4.Immunofluorescence showed that salvianolic acid B could significantly promote the expression of EMCN and HIF1 A.5.MicroCT analysis showed that in the rat model of osteoporotic fracture,three-dimensional imaging of bone mineral density,etc.,all confirmed that the drug group could promote fracture healing.ConclusionSalB,one of the extracts of Salvia Miltiorrhiza,may improve the local blood supply by promoting the expression of IncRNA-MALAT1 and thus accelerate the healing of osteoporotic fractures. |
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