蛇床子素对大鼠原代软骨细胞增殖的抑制作用  

Inhibition effect of osthole on proliferation of rat chondrocytes

作　　者：丁道芳[1,2] 韦宋谱[1,2] 李晓锋[3,4] 张晓钢[1,2] 詹红生[1,2] 段铁骊[1,2] 曹月龙[1,2] 

机构地区：[1]上海市中医药研究院骨伤科研究所,上海201203 [2]上海中医药大学曙光医院石氏伤科医学中心,上海201203 [3]上海中医药大学脊柱病研究所,上海200032 [4]上海中医药大学龙华医院,上海200032

出　　处：《中西医结合学报》2012年第12期1413-1418,共6页Journal of Chinese Integrative Medicine

基　　金：国家自然科学基金资助项目(No.81073114;81072830);上海市教育委员会创新项目(No.11YZ64);上海市科学技术委员会重点科技攻关项目(No.11DZ1973400)

摘　　要：目的:观察蛇床子素对大鼠软骨细胞增殖的影响。方法:分离出生24h内大鼠的软骨细胞,进行原代培养。细胞扩增至第2代时观察1周内细胞增殖情况,并用细胞免疫荧光法鉴定Ⅱ型前胶原基因(typeⅡprocollagen gene,Col2a1)的表达。将第2代软骨细胞分为对照组和不同浓度蛇床子素(6.25、12.5、25和50μmol/L)组。药物作用24和48h后,观察细胞增殖情况,蛋白质印迹法和聚合酶链反应法检测增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)和细胞周期素D1(cyclinD1)的表达。结果:第2代原代软骨细胞活力正常,Col2a1表达明显。蛇床子素可以抑制软骨细胞的增殖,并随浓度的增加,抑制作用愈明显。蛋白质印迹法和聚合酶链反应法检测发现,随着蛇床子素浓度的增加,软骨细胞中PCNA和cyclinD1的表达下降。结论:蛇床子素可能通过抑制PCNA和cyclinD1的表达发挥抑制软骨细胞增殖的作用。OBJECTIVE： To investigate the effects of osthole on chondrocyte proliferation in vitro. METHODS： Primary rat chondrocytes were isolated from the femoral head of newborn rats using collagenase digestion and cultured in Dulbecco＇ s modified Eagle＇ s medium. The proliferation of primary chondrocytes was assessed in second-passage cultures using cell counting kit-8 and the growth curve was drawn. Type ]] procollagen gene （Col2al） expression in chondrocytes was also identified using cell immunofluorescence assay. The second-passage chondrocytes were divided into five groups, including control group and osthole groups at 6.25, ]2.5, 25 and 50 μmol/L. The growth property of rat chondrocytes was observed after 24, 48 and 72 h of culture with osthole at corresponding dose. Both protein and mRNA expression of proliferating cell nuclear antigen （PCNA） and cyclin D1 was measured by Western blot and polymerase chain reaction methods. RESULTS. The second-passage chondrocytes were viable and showed Col2al expression in the cytoplasm. The proliferation of rat chondrocytes was inhibited by osthole in a dose-dependent manner. Meanwhile, there were significant decreases in both protein and mRNA expression of PCNA and cyclin D1 in the osthole groups compared with the control group. CONCLUSION： Osthole exhibits inhibitory effect on proliferation of rat chondrocytes by down-regulating PCNA and cyclin D1 expression.

关 键 词：植物提取物 蛇床子素 软骨细胞 细胞增殖 增殖细胞核抗原 细胞周期蛋白D1 大鼠 

分 类 号：R285.5[医药卫生—中药学]