| Objective: To investigate the influence of the tubulin destruction of chondrocyte metabolism in vitro.Methods: There were 8 New Zealand rabbits of 2 months old , and air embolism executed, obtained full thickness cartilage in both knees under sterile conditions. Then used 0.4% Pronase and 0.025%Ⅱcollagenase to digest the cartilage for the primary chondrocytes .Cultured the cells for 3 days after attachment, then divided them into control and experimental groups and control groups continue to culture using the primary medium and the experimental group in the medium with tubulin destructive agent- colchicine (final concentrationl:0.1μmol/l) . After 1 and 2 days , assayed early apoptosis of the two group cells using the phosphatidylserine (Annexin V). After 6 days,observed the cells morphological changes by HE staining. After 3,6,9 days measured the quantity of typeⅡcollagen, proteoglycan,and MMP-13 mRNA expression by real time quantitative fluorescent reverse transcription polymerase chain reaction (Real time RT-PCR).Result: after 2 days, early cell apoptosis rate of experimental group is higher than control group (P < 0.05). 6 days after dosing, observed experimental cell morphology is irregular and polygonal, hyperchromatic nucleus and split phase increased, matrix cell reduced. After 3,6 and 9 days, detected that the experimental groupⅡcollagen and proteoglycan mRNA expression is decreased comparing with the control group (P <0.05), the expression of MMP-13 mRNA is no significant changes in the control group after 3 days, which was significantly increased after 6 and 9 days(P <0.01).Conclusion: The destruction of tubulin can induce early apoptosis of chondrocytes, the synthesis and secretion of typeⅡcollagen and proteoglycan is reduced, the expression of MMP-13 is increased, cartilage cell degenerated, and the cartilage cells of the synthesis and catabolism environmental balance is disorder. Finally,all of them would make a serious decline in biological functions.
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