目的：探讨跳骨片水提物对体外大鼠关节软骨细胞活性的影响。方法：4周龄SPF级SD大鼠5只，采用机械-Ⅱ型胶原酶消化法获取膝关节软骨细胞，倒置相差显微镜观察软骨细胞的形态结构，Ⅱ型胶原免疫组化染色鉴定软骨细胞。0，25，50，100μg·mL-1跳骨片水提物对第3代软骨细胞，分别干预24，48，72 h后，MTT法检测细胞活性。结果：①Ⅱ型胶原免疫组化染色后软骨细胞胞浆呈棕黄色，为阳性；②以0μg·mL-1跳骨片水提物干预的软骨细胞作为对照组，干预24 h后，50μg·mL-1组软骨细胞活性明显高于对照组（P <0.05）；干预48h后，25，50μg·mL-1组明显高于对照组（P <0.05）；干预72h后，25，50，100μg·mL-1组与对照组比较，差异无统计学意义（P >0.05）。结论：跳骨片水提物在25，50，100μg·mL-1时均可以促进软骨细胞的增殖，具有剂量和时间依赖性，其中50μg·mL-1干预24 h后最为明显。%Objective:To investigate the effects of aqueous extract of Tiaogu Tablet(跳骨片)on the activity of rats articular chondrocytes in vitro.Methods:Articular cartilage cells were got from ifve 4-week-old SD rats of SPF grade by the mechanical typeⅡcollagenase digestion method and their morphology was observed under inverted microscope.The cartilage cells were identiifed by the typeⅡcollagen immunohistochemical staining. After the third generation of chondrocytes were intervened with 0,25,50,100 μg·mL-1 of aqueous extract of Tiaogu Tablet respectively after 24,48,72 h,their activity was tested with MTT method.Results:① Chondrocyte cytoplasm was brown and positive after staining;②The activity of chondrocytes intervened with 50 μg·mL-1 of aqueous extract of Tiaogu Tablet after 24 h was significantly higher than the control group with 0 μg·mL-1 of aqueous extract of Tiaogu Tablet(P< 0.05).After intervention for 48 h,the activity of chondrocytes in groups intervened with 25,50 μg·mL-1 of extract was significantly higher than the control group (P< 0.05).After 72 h intervention,the differences between groups intervened respectively with 25,50, 100 μg·mL-1 of aqueous extract of Tiaogu Tablet and the control group were not statistically signiifcant(P> 0.05). Conclusion:Aqueous extract of Tiaogu Tablet of 25,50,100 μg·mL-1 can promote the proliferation of chondrocytes,depending on dose and time,of which 50 μg·mL-1makes it obvious after 24-hour intervention.
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