跳骨片水提物对体外大鼠关节软骨细胞活性的影响

郑文伟+马玉环+林平冬+陈后煌+邵翔+陈达+李西海+叶蕻芝

【摘 要】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。0，25，50，100 μg·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0 μg·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，50 μg·mL-1组软骨细胞活性明显高于对照组（P < 0.05）；干预48 h后，25，50 μg·mL-1组明显高于对照组（P < 0.05）；干预

72 h后，25，50，100 μg·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25，50，100 μg·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50 μg·mL-1干预24 h后最为明显。

【关键词】 骨关节炎；跳骨片水提物；MTT法；软骨细胞；大鼠

Effects of Aqueous Extract of Tiaogu Tablet（跳骨片）on the Activity of Rats Articular Chondrocytes in Vitro

ZHENG Wen-wei，MA Yu-huan，LIN Ping-dong，CHEN Hou-huang，SHAO Xiang，CHEN Da，LI Xi-hai，

YE Hong-zhi

【ABSTRACT】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 five 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 identified 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 significant（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.

【Keywords】 osteoarthritis；aqueous

extract of Tiaogu Tablet（跳骨片）；MTT method；

chondrocyte；rats

骨關节炎（osteoarthritis，OA）是一种以关节软骨退变为主的慢性、进行性骨关节疾病[1-2]。OA始发部位在软骨，以中老年人群最为常见。软骨退变是多因素综合作用的结果，软骨细胞作为软骨中的唯一细胞及软骨破坏过程的靶细胞，其结构的破坏及功能丧失在此过程中起着重要作用[3-4]。因此，采取保护软骨细胞结构及调节软骨细胞功能可能是防治OA重要途径之一。目前治疗OA的方法主要以对症治疗为主，缺乏特效疗法。中医药在治疗OA上具有多靶点、多方向、安全可靠、经济等特点[5]。跳骨片具有活血舒筋、消肿定痛等功效[6]，但目前其在治疗OA方面的研究甚少，因此初步研究跳骨片水提物对体外关节软骨细胞活性的影响，有利于为OA的治疗及跳骨片开发利用提供一定理论依据。……