The Study Of Blocking The SDF-1/CXCR4Signaling Pathway To Regulate The Degeneration Of Articular Cartilage With AMD3100

Objective:To explore the SDF-1/CXCR4signal pathway mechanism in primary osteoarthritis of the Hartley guinea pig by dynamic observing the degenerative change and the morphological change and the relative blood and cartilage biochemical indexes in Hartley guinea pig articular cartilage,On this basis.to explore the the feasibility that blocking SDF-1/CXCR4signal pathway with AMD3100to delay the degenetation of cartilage and find the mechanism of AMD3100.Methods:1.40Hartley guinea pigs were divided into two large groups (each n=20):Female group and Male group. and each group was devided into3,6.9.12months sub group.5in each sub group.The tibial plateau cartilage tissue were taken for histology examination (HE and Safranin-O staining) and gotten Mankin histological score; ELISA was used to evaluate of the levels of SDF-1and estradiol in serum; Reverse transcription PCR was used to test the expression of MMP-3,9,13, CXCR4, Aggrecan, type II collagen in the cartilage tissue;Western Blot was taken to test the type II collagen of the cartilage tissue; SPSS17.0was used for date analysis.2.36male Hartley guinea pigs(6-month-old) were divided randomly into three groups: the experimental group(Group A), the experimental control group(Group B), the blank control group (Group C)(each n=12), Group A was subcutaneously implanted Alzet micro-pump, which contained and pumped into the diluted PBS with AMD3100(the concentration of AMD3100was180ug/ml.d).Group B was subcutaneously implanted Alzet micro-pump, which contained and pumped the diluted PBS.Group C was received no treatment. ELISA was used to test of the levels of SDF-1in serum after 2,4,8,10,12weeks;And after12weeks,The Hartley guines pigs were sacrificed and the tibial plateau cartilage tissue was taken for histology examination (HE and Safranin-O staining) and gotten Mankin histological score and the tibial plateau cartilage tissue was also taken for IL-1, TNF-a immunohistochemistry; ELISA was used to determine of the levels of SDF-1in serum; Real Time Quantitative PCRwas used to test the expression of MMP-3,9,13,CXCR4,Aggrecan,type II collagen mRNA in the cartilage tissue;Western Blot was taken to test the type II collagen of the cartilage tissue;SPSS17.0was used for date analysis.Results:1.1-1Histology Test (HE staining and Safranin staining):With the month of the Hartley guinea pig increase,the degeneration of the cartilage tissue increased gradually,the Mankin histological score was increased gradually, the difference was statistically significant(P<0.05).1-2ELISA examination:The levels of SDF-1and estradiol in the different age groups of female was statistically significant (P<0.05); The levels of SDF-1in the different age groups of male was statistically significant (P <0.05); The levels of estradiol in the different age groups of male was no significant (P>0.05).1-3Reverse transcription PCR: With the increase in months of age,the expression of CXCR4, MMP-3,9,13cartilaginous tissue increased gradually,The comparison each month group was significantly different (P<0.05); the expression of proteoglycan of cartilage tissue, type II collagen mRNA decreased gradually, The comparison each month group was significantly different (P<0.05);1-4Western Blot: With the increase of month, the level of type II collagen decreased gradually, The comparison each month group was significantly different (P<0.05);1-5The levels of estradiol is positively correlated with the levels of SDF-1in the different age groups of female,and positively correlated with the expression of CXCR4.2.2-1Histology Test (HE staining and Safranin staining):By the Mankin scores, the experimental group had lower than the experimental control group and blank control group,with thedifference statistically significant (P<0.05);2-2Immunohistochemistry: The expression of IL-1, TNF-a was weakly positive in the experimental group,and the expression of IL-1.TNF-a was positive or strongly positive in the experimental control group and blank control group.2-3ELISA:In the experimental group.The levels of SDF-1in serum were significantly lower than the experimental control group and blank control group,the difference was statistically significant (P<0.05).2-4Real Time Quantitative PCR:The expression of MMP-3,9,13mRNA was lower in experimental group compared with experimental control group and blank control group, the difference was statistically significant (P<0.05); The expression of type II collagen, aggrecan mRNA in experimental group were higher than the experimental control group and the blank control group,the difference was statistically significant (P<0.05);2-5Western Blot:The level of type II collagen in experimental group was higher than the experimental control group and the blank control group, the difference was statistically significant (P<0.05).Conclusion:1. With the increase of months, the osteoarthritis occured in Hartley guinea pigs in knee joint cartilage,the3-month-old guinea pig knee joint was normal;6-month-old guinea pig knee joint got degenerately;9-month-old guinea pig could be the typical primary osteoarthritis of pathological changes;12-month-old guinea pig knee joint degenerated seriously.2.There were SDF-1/CXCR4signal pathway in Hartley guinea pigs' osteoarthritis, and with the increase of the months age can be induced the progress of signaling pathway.3.The levels of estradiol was positively correlated with SDF-1/CXCR4signal pathway in female Hartley guinea pig.4.AMD3100clould delay the articular cartilage degeneration by blocking the SDF-1/CXCR4signal pathway.5.AMD3100clould reduce the expression and secretion of MMP-3,9,13,and the degradation of type II collagen and aggrecan in vivo cartilage.