Experimental Study On The Synergistic Effect Of Curcumin And Magnesium Sulfate On The Reverse Differentiation Of Inflammatory Chondrocytes

Background and ObjectiveOsteoarthritis is a common cause of pain and disability in the elderly.As the population ages and people's living standards increase,the incidence increases gradually.The underlying causes are obesity,inappropriate mechanical stress,inflammatory transmitters and biochemical factors.The synergy.The treatment of early osteoarthritis includes oral medication,intra-articular injection,autologous chondrocyte transplantation through minimally invasive surgery,osteochondral allograft bone implantation,and arthroscopic microfracture treatment.At present,the research shows that the curative effect is acceptable for a period of time,and it has been found that it has limited curative effect on cartilage degradation,regeneration and repair,and finally needs to be solved by joint replacement.However,surgery brings a large economic burden and more mental and psychological stress to patients,so early drug treatment to solve the degeneration of osteoarthritis still has important clinical significance.Curcumin is extracted from the rhizome of Zingiberaceae,and its main component is behenyl methane.It has been proven to have anti-oxidation,inflammation,antibacterial,anti-tumor,neuroprotective,cardioprotective,radiation protection and inhibition of joint retreat.The role of change.Magnesium,as one of the essential elements of life,plays an indispensable role in the human body.Magnesium sulfate(MgSO4)is often used in clinical medicine to treat diseases such as warts and pre-eclampsia,and it also relieves joint pain and inhibits joint degeneration.There is currently no research on the effects of curcumin and magnesium sulfate on inflammatory chondrocytes and its mechanisms.This study will collect inflammatory cartilage from patients with clinical osteoarthritis,and perform isolation and original culture.The expression of inflammatory chondrocyte protein and gene level was detected by immunofluorescence staining and quantitative PCR.In vitro 3D culture chondrocyte differentiation-specific staining and TUNEL cell apoptosis staining were performed to detect the degree of inflammatory cartilage reverse differentiation.To explore the effect of curcumin and MgSO4 on the reverse differentiation of inflammatory chondrocytes and its related mechanisms.For the early clinical treatment of inflammatory chondrocyte degeneration,provide new drug treatment ideas and methods.Method1.Isolation and culture of proto-inflammatory chondrocytes,MTS cell proliferation assay to detect the proliferation of progenitor inflammatory chondrocytes in the second and third generation cells under monolayer culture conditions.2.Combination of curcumin and MgSO4 can promote the expression of proteoglycan,a cartilage differentiation marker,and chondrocytes can secrete more cartilage matrix.3.Cell slide,immunofluorescence staining and QPCR quantitative detection of monolayer cell culture,4 groups of cells into cartilage marker protein and gene level expression.4.The cells in 4 groups were induced to differentiate in vitro by 3D for 18 days,and 4 groups of cell pellets were harvested.After measuring the cell mass size of each group,paraffin-embedded sections,Alcian blue,Safranin O staining and TUNEL cell apoptosis detection(color development method),the area of the cartilage differentiation positive area and the area of apoptosis-positive area of each group.5.All trials were repeated 3 times or more.Data results were expressed by x±s.Data processing and statistical analysis were performed using SPSS 22.0 software.One-way analysis of variance was used for comparison between groups.P<0.05 was considered statistically significant.Results1.MTS cell proliferation experiment results show that the second generation of inflammatory chondrocytes has a higher rate of early proliferation and a lower degree of differentiation.2.Cell slides and immunofluorescence staining showed that the expression levels of type ? collagen in the chondrogenic differentiation markers were higher in the combination therapy group,curcumin group and MgSO4 group than in the inflammatory chondrocytes group(P<0.01).3.QPCR quantitative results showed that the expression levels of type ? collagen,proteoglycan and SOX9 mRNA in the combined treatment group were higher than those in the curcumin group or MgSO4 group(P<0.05).The expression level of the gene SOX9 was better in the MgSO4 group than in the curcumin group(P<0.01).4.After in vitro 3D-induced differentiation,the cell size,cartilage reverse differentiation,Alcian blue,and Safranin-positive positive staining were significantly higher in the combination group than in the other 3 groups(P<0.01).The results of TUNEL apoptosis staining showed that the area of positive area of the combined treatment group and MgSO4 group was significantly smaller than the other two groups(P<0.01).ConclusionThe combined application of curcumin and MgSO4 has a synergistic effect on promoting the reverse differentiation of inflammatory chondrocytes,which may be achieved by inducing the differentiation of inflammatory chondrocytes into normal chondrocytes,inducing the up-regulation of SOX9 gene of inflammatory chondrocytes by MgSO4,and inhibiting the apoptosis of inflammatory chondrocytes.