The Study Of Targeted Blocking The SDF-1/CXCR4Signaling Pathway To Regulate The Degeneration Of Articular Cartilage With T140

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 T140to delay the degenetation of cartilage and find the mechanism of T140.Methods: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 T140(the concentration of T140was180ug/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 after2,4,6,8,10,12weeks;And after12weeks,The Hartley guines pigs were sacrificed, glenoid,head of humerus,condyles of humerus,femoral head,femoral condyle,patella,tibial plateau,ankle joint. 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-1in serum; Real Time Quantitative PCR was used to test the expression of MMP-3,9,13,Aggrecan,type Ⅱ collagen mRNA in the cartilage tissue;Western Blot was taken to test the type II collagen of the cartilage tissue;SPSS19.0was used for date analysis.Results:1Histology Test (HE staining and Safranin staining):By the Mankin scores, the experimental group was lower than the experimental control group and blank control group,with thedifference statistically significant (P<0.05);2Immunohistochemistry:Positive cells in experimental group compared with the Experimental control group and blank control was low (P<0.05)。The experiment between the control group and blank control group of IL-1and the expression of TNF-a had no diffeence.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).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);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).Conclusions:1.T140could targeted blocking the SDF-1/CXCR4signaling pathway, on the body in vivo most weight-bearing joints,such as the shoulder, elbow, hip, knee, ankle and others.and it could delay the degeneration of articular cartilage.2. T140could block the SDF-1/CXCR4signal pathway and reduce the expression and secretion of MMP-3,9,13in vivo to delay the articular cartilage degeneration. 3. T140could block the SDF-1/CXCR4signal pathway and reduce the degradation of type Ⅱ collagen and aggrecan in vivo to delay the articular cartilage degeneration.