| Objective:To investgate the effects of potassium and chloride channel blockers on The glycosaminoglycan(GAG) and collagen type II synthesis of rabbit articular chondrocytes in vitro.Methods:2 months New Zealand rabbits were killed and their knee joint were taken out under aseptic condition. The chondrocytes were isolated by enzymolysis method and cultivated in 24-well plates (seeded 5×104 cells per well).Then the chondrocytes were divided into three groups randomly after cultured 2 days when the cells were adherent. The control group was cultured by DMEM/F12 while the potassium channel blockon group(4-AP group)was cultured in the media contained 1mmol/L 4-aminopyridine (4-AP), which can lead to the blockon of the potassium channel,and the chloride channel blockon group(SITS group)was cultured in the media contained 1mmol/L 4-acetamido-4-isothiocyano-2,2-disulfonic acid stilbene(SITS), which can lead to the blockon of the chloride channel.The GAG synthesis were measured by Alician Blue method and collagen typeⅡsynthesis were estimated by the ELISA in the media when medium was change after3-,6-and 9-day (n=12)Result:Compared with the control group.the GAG content of 4-AP group has significant decrease at 3th day(P<0.05),but has no significant difference at 6th and 9th day (p>0.05),while the collagen typeⅡcontent has significant increased at 3th and 6th day (P<0.05), and has decrease tendency at 9th day. but have no significant difference.The GAG content of SITS group has significant increased at 3th,6th and 9th day (P<0.05).while the collagen typeⅡcontent has significant increased at 3th and 6th day (P<0.05),and has increased tendency at 9th day, but have no significant difference.Conclusion:The blockon of potassium and chloride channel can increase the GAG and collagen typeⅡsynthesis of chondrocytes in vitro, especially when block chloride channel,the GAG synthesis was significant. |
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