Effects Of Eicosapentaenoic Acid On The Vitality And Adipogenic Differentiation Ability Of Bone Marrow Mesenchymal Stem Cells

Background:The prevalence of osteoporosis(OP)is increasing year by year,and its incidence in my country is closely followed by cardiovascular disease and diabetes,ranking third among chronic diseases,seriously endangering the health of middle-aged and elderly people.Drugs for the treatment of osteoporosis have been greatly developed in recent years,but there are still some problems to be solved in these drugs,and it is of certain significance to discover new methods.Most of the current research proves that there is an inverse relationship between bone marrow fat mass and bone mass.In the bone marrow,bone marrow mesenchymal stem cells(BMMSCs)are responsible for the differentiation into bone marrow adipocytes and osteoblasts.There is a lot of evidence that inhibiting the adipogenic differentiation of BMMSCs may be one of the potential treatments of osteoporosis.Omega-3 long-chain polyunsaturated fatty acids are considered to be highly biologically active in our body.Among them,Eicosapentaenoic Acid(EPA)is ubiquitous in marine animals and plants,and it has been found to have potential applications in many diseases,including bone health.Purpose:To explore the effects of different concentrations of EPA on the vitality and adipogenic differentiation ability of mouse BMMSCs,and to clarify the therapeutic and preventive significance of EPA on osteoporosis at the cellular level.Methods:1.Different concentrations of EPA were added to the BMMSCs of C57BL/6J mice to co-culture for 24 hours,and compared with the blank control group,the effect on the ability of BMMSCs was detected.2.Oil red O staining was used to observe the effect of different concentrations of EPA on the number of BMMSCs differentiated into adipocytes.3.The size and state of lipid droplets in adipocytes differentiated from BMMSCs by different concentrations of EPA was observed by optical microscope.4.RT-qPCR was used to detect the effect of different concentrations of EPA on the gene expression of peroxisome-activated receptor y(PPARy)and CCAAT enhancer-binding protein a(C/EBPa)during adipogenic differentiation of BMMSCs.5.The effect of different concentrations of EPA on the expression of peroxisome-activated receptor y protein(PPARy)and adipose tissue fatty acid binding protein(FABP4)during the adipogenic differentiation of BMMSCs was detected by Western-Blot method.Results:1.Concentrations of 30 μM and 60 μM can significantly promote the proliferation of BMMSCs after 24h treatment,with statistical differences(P<0.001);150 μ,200 μM,and 300 μM have obvious inhibitory effects on the activity of BMMSCs,with statistical differences.(P<0.001).2.On the 7th day of induction of adipogenic differentiation of BMMSCs,there was no significant difference in the number of adipocytes between 60μM and the blank control group,while the number of adipocytes at 100μM concentration was significantly reduced compared with the blank control group.3.Under microscope observation,EPA inhibited the fusion of lipid droplets in adipocytes differentiated from BMMSCs in a concentration-dependent manner.4.On the 7th day of adipogenic induction of BMMSCs,the expression of PPARy gene was significantly inhibited in the 60μM group compared with the blank control group(P<0.05),while the expression of C/EBPa gene had no significant difference;the 100μM group and the control group had no significant difference.Compared with the control group,the expression of PPARy(P<0.01)and C/EBPα gene(P<0.05)were significantly inhibited.Conclusion:In this study,we concluded that EPA can inhibit the adipogenic differentiation of BMMSCs into adipocytes and the fusion of lipid droplets in adipocytes,and provide new evidence for the development of EPA on osteoporosis.