Study On The Effect Of Glucosamine Sulfate On Chondrocyte Degeneration Of Cervical Cartilage Endplate

Objective Study on the effect of glucosamine sulfate on chondrocyte degeneration of cervical cartilage endplate Methods 1.According to the Pfirrmann intervertebral disc degeneration rating(quasi I-V)5 type of human cervical intervertebral disc cartilage endplate,the cartilage cells in the cartilage endplate of cervical intervertebral disc were obtained by enzymolysis and digestion,and identification of chondrocytes on cells by toluidine blue staining.The growth of chondrocytes was observed by phase contrast microscope and the growth curve was plotted by colorimetric method.2.Cartilage cells were successfully isolated from cartilage endplates and cultured in vitro for 3 generations.A typical group of cells in each group was labeled as A,B,C,D,E,five group,Each group was divided into the control group and the experimental group(experimental group was 10?M,100?M,1000?M concentration of glucosamine sulfate)intervened for 72 hours,the expression of type II collagen,polyproteoglycan(Aggrecan),MMP-3 and MMP-13 genes were detected by q-PCR.Results ??The enzyme digestion method was used to obtain a normal form,to obtain a large number of pure chondrocytes,and to be able to proliferate in vitro.? ? 1 ? Gene expression of Aggrecan by GS(1)Group A : The GS concentration was 10?M expression down-regulation of Aggrecan gene,The GS concentration was 100 ? 1000?M expression up-regulation of Aggrecan gene;Group B?C:The GS concentration was 10?M expression up-regulation of Aggrecangene,The GS concentration was 100 ? 1000?M expression down-regulation of Aggrecan gene;Group D: The GS concentration was 10? 100 ? ? expression up-regulation of Aggrecan gene,The GS concentration was 1000?M expression up-regulation of Aggrecan gene;Group E:The GS concentration was 10?100 ??expression up-regulation of Aggrecan gene,The GS concentration was 1000?M expression up-regulation of Aggrecan gene;(2)GS is 0?10?M,the expression of B,C,D,E and A compared with the up regulation of Aggrecan gene;GS is1000?M,the expression of B,D,E and A compared the up-regulation of Aggrecan gene,the expression of C and A compared with the down-regulation of Aggrecan gene.2? Gene expression of type II collagen by GS(1)Group A ? B : The GS concentration was 10 ? 100?M,up regulate the expression of type II collagen gene,The GS concentration was 1000?M,up regulate the expression of type II collagen gene;Group C?D?E:The GS concentration was 10 ?100?1000?M,up regulate the expression of type II collagen gene;(2)GS at 0? 10?M,the expression of B,C,D to A compared to the upregulation of collagen type II gene expression of E,compared with the down-regulation of collagen II gene,GS is1000?M,the expression of B,D,E and A compared to the upregulation of type II collagen gene,expression of C and A reduced compared to type II collagen gene.3?Expression of MMP-3 gene by GS(1)Group A?D:MMP-3 gene expression was up-regulated when the concentration of GS was 10,100,and1000?M;Group B:MMP-3 gene expression was down-regulated when the concentration of GS was 10,100?M;MMP-3 gene expression was up-regulated when the concentration of GS was 1000?M;Group C: MMP-3 gene expression was up-regulated when the concentration of GS was 10,100?M,MMP-3 gene expression was down-regulated when the concentration of GS was 1000?M;Group E:MMP-3 gene expression was up-regulated when the concentration of GS was 10,1000?M;MMP-3 gene expression was down-regulated when the concentration of GS was 100?M;(2)The concentration of GS 10 ?M: B,C,D,E up-regulated the expression of MMP-13 gene compared with A;the concentration of GS was 100 ?M: B,E compared with A up-regulated the expression of MMP-13 gene,C,D and Compared with A,the expression of MMP-13 gene was down-regulated;the concentration of GS was 1000 ?M: B,D compared with A up-regulated the expression of MMP-13 gene;C,E compared with A down-regulated the expression of MMP-13 gene.4?Expression of MMP-13 gene by GS(1)Group A?B?C:MMP-13 gene expression was up-regulated when the concentration of GS was 10,100,and1000?M;Group D : MMP-13 gene expression was down-regulated when the concentration of GS was 10,100,?M,MMP-13 gene expression was up-regulated when the concentration of GS was 10,100,?M;(2)GS reduced the expression of MMP-13 gene at 0 and 10 ?M:B,C,D,E compared with A;when GS was 100 ?M,B,C,D,and MMP-3 gene were down-regulated compared with A,E.A compared with down-regulated MMP-3 gene expression;GS is 1000??,C,D,E and A compared with down-regulation of MMP-3 gene expression,B compared with A down-regulated MMP-3 gene expression.Conclusion 1?Gene expression of Aggrecan by GS:Aggrecan gene expression in cartilage cartilage endplate chondrocytes of I-V degenerated cervical cartilage was up-regulated at 10 ?M;Aggrecan gene expression in cartilage cartilage endplate chondrocytes of I-V degenerated cervical cartilage was down-regulated at100?1000 ?M.2? Gene expression of type II collagen by GS: type II collagen in cartilage cartilage endplate chondrocytes of I-V degenerated cervical cartilage wasup-regulated at 10 ? 100 ?M,;type II collagen in cartilage cartilage endplate chondrocytes of I-V degenerated cervical cartilage was up-regulated at 1000?M,;3 ? Expression of MMP-3 gene by GS : GS up-regulated the expression of MMP-3 gene in cartilage of I-III-,IV-,and V-type degenerative cartilage endplates,at the concentration of 10 ?M,and down-regulated the expression of MMP-3 gene in type II degenerated cervical cartilage end-plate chondrocytes.GS up-regulated the expression of MMP-3 gene in cartilage of I-III-and V-type degenerative cartilage endplates,at the concentration of 100 ?M,and down-regulated the expression of MMP-3 gene in type II and V degenerated cervical cartilage end-plate chondrocytes.GS up-regulated the expression of MMP-3 gene in cartilage of I-II-IV and V-type degenerative cartilage endplates,at the concentration of 1000?M,and down-regulated the expression of MMP-3gene in type III degenerated cervical cartilage end-plate chondrocytes.4 ? Expression of MMP-3 gene by GS : GS up-regulated the expression of MMP-13 gene in cartilage of I-II and III-type degenerative cartilage endplates,at the concentration of 10 ?M,and down-regulated the expression of MMP-13 gene in type IV and V-type degenerated cervical cartilage end-plate chondrocytes.GS up-regulated the expression of MMP-13 gene in cartilage of I and II-type degenerative cartilage endplates,at the concentration of 100 ?M,and down-regulated the expression of MMP-13 gene in type III-IV and V-type degenerated cervical cartilage end-plate chondrocytes.GS up-regulated the expression of MMP-13 gene in cartilage of I-II-and IV-type degenerative cartilage endplates,at the concentration of 1000 ?M,and down-regulated the expression of MMP-13 gene in type III and V-type degenerated cervical cartilage end-plate chondrocytes.It shows that different concentrations of GS have certain protective effects on some degenerative types of cervical cartilage endplates,but the mechanism is more complex.