The Research Of Blocking SDF-1/CXCR4 Signaling Pathway To Delay The Degeneration Of Articular Cartilage With T140

[Objective]:To study the role of stromal cells derived factor-1 (SDF-1)signaling pathway in articular cartilage degeneration, and explore the feasibility that blocking SDF-1/CXCR4 signaling pathway with T140 can delay the degenetation of articular cartilage and its mechanism, thus provide theoretical basis for the targeted therapy of osteoarthritis(OA).[Methods]:The cartilage tissues were acquired in size of 3×3×1mm from the knee cartilage surface (Mankin score of 0 or 1).12 cases which diagnosed as OA were taken from total knee replacement surgery, named as OA cartilage group. Another 12 normal cases were taken from traumatic amputation, named as normal cartilage group. The cartilage Tissues were cultured in the nutrient solution containing of SDF-1 100ng/ml, added T140, MAB310, MAB002 1000nM respectively, cultured in 37℃CO2 incubator for 2 or 4 days. The cartilage tissues were classified into 4 groups:(1) the experimental group:T140+SDF-1, (2) the experimental control group1: MAB310+SDF-1, (3) the experimental control group 2:MAB002+SDF-1, (4) the blank control group:nothing+SDF-1。The cartilage tissues were cultured for 2 or 4 days. Then the cartilage tissues and nutrient solution were taken to following testings:1. Histologic examination (HE and Safranin-O stainning); 2. RT-PCR was taken to test the expression of MMP-3,-9,-13, type II collagen, aggrecan mRNA in the cartilage tissues; 3. Western blotting was taken to test the type II collagen of the cartilage tissues; 4. ELISA was used to measure the capacity of MMP-3,-9,-13 in the culture medium. SPSS 17.0 were used for data analysis.[Results]:(1) Histologic examination (HE and Safranin-O stainning):Compared to the experimental control group and the blank control group cartilage, there were more cartilage cells in the experimental group cartilage and cytoplasmic staining was more even; (2) RT-PCR and Western blotting:In the experimental group the experimental control group and the blank control group cartilage, the expression of MMP-3,-9,-13 was lower and the expression of typeⅡcollagen and aggrecan was higher than the experimental control group and the blank control group Cartilage, the difference was statistically significant (P<0.05); (3) ELISA:In the culture medium of experimental group, the capacity of MMP-3,-9,-13 was lower than the experimental control group and blank control group, the difference was statistically significant (P<0.05)[Conclusions]:1. SDF-1 could induce the articular cartilage degeneration in vitro culture.2. T140 could delay the articular cartilage degeneration by blocking the SDF-1/CXCR4 signaling pathways.3. T140 could delay, but could't reverse OA articular cartilage into nomal form in short-term.