鹿茸多肽促进软骨细胞增殖机制的初步探讨  

The mechanism of antler polypeptides promoting chondrocyte proliferation

作　　者：林建华[1] 邓凌霄[1] 吴朝阳[1] 陈雷[1] 

机构地区：[1]福建医科大学附属第一医院骨科,福州350005

出　　处：《中华创伤骨科杂志》2007年第12期1161-1164,共4页Chinese Journal of Orthopaedic Trauma

基　　金：福建省科技厅重点项目（2004Y018）

摘　　要：目的探讨鹿茸多肽（PAP）促进软骨细胞增殖的机制。方法以12．5、25．0、50．0、100．0μg／mL不同浓度的PAP刺激软骨细胞，倒置相差显微镜观察软骨细胞生长情况，噻唑兰法（MTT）检测软骨细胞的生长增殖能力；以6、12、24、48μg／mL不同浓度酪氨酸受体激酶阻断剂Genis—tein作用于受PAP刺激的软骨细胞，用MTT和流式细胞仪观察Genistein对PAP作用的影响；免疫组化SABC法检测核增殖抗原cyclinA在PAP和PAP＋Genistein作用下的表达变化，RT—PCR法检测c-fosmRNA表达。结果PAP作用下c-fos mRNA与cyclinA表达增加，s期的细胞比例增高，从6．4％增加到35．2％；Genistein作用后PAP的促增殖作用受到明显的抑制，s期的细胞降到4％，c-fos mRNA与cyclinA表达亦减少；单纯用Genistein不影响软骨细胞生长。结论PAP可能通过酪氨酸受体蛋白激酶介导，作用下游的信号，引发立早基因c-fos表达，促进cyclinA的表达，引起更多的细胞进入有丝分裂期，从而促进软骨细胞增殖。Objective To study the mechanism of antler polypeptides （PAP） promoting chondrocyte proliferation. Methods Cultured rabbit chondrocytes were stimulated by various concentrations of PAP （ 12.5 μg/mL, 25.0μg/mL, 50. 0 μg/mL, 100. 0μg/mL） and by PAP （50 μg/mL） plus various concentrations of genistein, a tyrosine kinase inhibitor （6 μg/mL, 12 μg/mL, 24 μg/mL, 48 μg/mL） respectively. Cell proliferations were observed by the inversion phase contrast microscopy and determined by MTT assay and flow cytometry. SABC techniques were applied for cyclin A immunocytochemical staining. The expression of c-fos mRNA was assessed by RT-PCR. Results PAP increased proliferation percentage and cells at S stage of cell cycle, up-regulating the percentage of positive staining cyclin A cells and expression of e-fos mRNA. But Genistein suppressed the proliferation by decreasing the percentage of positive staining cyclin A cells and expression of e-fos mRNA at S stage. Both of the two substances showed a dose-dependent manner. Conclusion PAP stimulates the proliferation of chondrocytes by inducing more S-stage entry via tyrosine kinase pathway, which induces the expression of proto-oncogenes c-fos responsible for modulating cyclin A.

关 键 词：软骨 增殖 周期蛋白 生长物质 

分 类 号：R686[医药卫生—骨科学]