The Study On Delaying The Degeneration Of Articular Cartilage By Targeted Blocking SDF-1/CXCR4 Signaling Pathway With Three Antagonists In Vivo

Objective:To explore the possibility and mechanism through targeted blocking SDF-1/CXCR4 signaling Pathway with three antagonists TN14003,T140,AMD3100 in vivo,and to investigate the function of three antagonists to delay degeneration process of articular cartilage.Methods:96 male Duncan-Hartley guinea pigs(6 months old)were divided into group A,B,C and D randomly.Alzet trace pump was implanted in the back subcutaneous tissue of pigs in group A and TN14003 with concentration of 180ug/ml was pumped every day.Alzet trace pump was implanted in the back subcutaneous tissue of pigs in group B and T140 with concentration of 180ug/ml was pumped every day.Alzet trace pump was implanted in the back subcutaneous tissue of pigs in group C and AMD3100 with concentration of 180ug/r(?)was pumped every day.Hartley guinea pigs in group D received no treatment.In the 2nd,4th,6,8th,10th,12th week,the levels of SDF-1 in serum were quantified with ELIS A.All the pigs were sacrificed after 12-week conventional breeding and the cartilage of both knees was removed immediately,Their cartilage tissue was taken for histology examination(HE and Safranin-O staining)and gotten Mankin histological score.Cartilage tissue was also taken for IL-1,TNF-a immunohistochemistry;ELISA was used to determine of the levels of SDF-1 in serum;Real Time Quantitative PCR was used to test the expression of MMP-3,9,13,Aggrecan,type ? collagen mRNA in the cartilage tissue;Western Blot was taken to test the type ? collagen of the cartilage tissue;SPSS19.0 was used for data analysis.Results:(1)Histology Test(HE staining and Safranin staining):the Mankin scores,results showed that groupA<groupB<groupC<groupD,with the difference statistically significant(P<0.05).2 Immunohistochemistry:Positive cells of the expression of IL-1 and TNF-a showed A<B<C<D,and there was statistically significant(P<0.05).3 ELISA:the serum levels of SDF-1 of the group A,B,C decreased gradually and group A showed significantly,but increased in group D.At the same time,the serum levels of SDF-1 of the group A were significantly lower than that of group B,those of group B were significantly lower than that of group C,which was significantly lower than that of group D,and there was statistically significant(P<0.05).In group A,The levels of SDF-1 in serum were significantly lower than that of groupB,groupB was lower than that of groupC,groupC was lower than that of groupD,and there was statistically significant(P<0.05).4 Real Time Quantitative PCR:The mRNA levels of MMPs in group A were significantly lower than the group B,those of group B were significantly lower than group C,which was significantly lower than group D,and there was statistically significant(P<0.05);The mRNA levels of type II collagen,aggrecan in group A were significantly more than group B,those of group B were significantly more than group C,which was significantly more than group D,and there was statistically significant(P<0.05).5 Western Blot:The collagen II expression in group A were significantly more than in group B,those of group B were significantly more than group C,which was significantly more than group D,and there was statistically significant(P<0.05).Conclusions:1.TN14003,T140,AMD3100 could targeted blocking the SDF-1/CXCR4 signaling pathway,and they could delay the degeneration of articular cartilage.2.Three antagonists could block the SDF-1/CXCR4 signal pathway and reduce the expression and secretion of MMP-3,9,13 in vivo to delay the articular cartilage degeneration.3.Three antagonists could block the SDF-1/CXCR4 signal pathway and reduce the degradation of the collagen ? and aggrecan in vivo to delay the articular cartilage degeneration.4.TN14003 had better effect on preventing OA cartilage degeneration by blocking the SDF-1/CXCR4 signal pathway than T140 and AMD3100.