正钒酸钠对椎间盘软骨细胞功能的抑制作用  

Effects of Na3Vo4 on the chondrocyte in the intervertebral disc

作　　者：王拥军[1,2] 石继祥[1,2] 施杞[1,2] 周泉[1,2] 唐占英[1,2] 卞琴[1,2] 周重建[1,2] 

机构地区：[1]上海中医药大学脊柱病研究所,200032 [2]上海中医药大学附属龙华医院骨伤科

出　　处：《中华骨科杂志》2007年第3期212-216,共5页Chinese Journal of Orthopaedics

基　　金：国家杰出青年科学基金资助项目（30625043）;国家自然科学基金重点资助项目（30330700）;国家自然科学基金资助项目（30371794、30572398）;上海市科技“启明星”跟踪计划（05QMH1412）;上海市医学重点学科建设项目（05Ⅲ027） 【致谢】感谢上海瑞金医院伤骨科研究所邓廉夫教授、冯伟副教授、许福平老师给本研究的帮助.

摘　　要：目的探讨正钒酸钠（Sodium Orthovanadate，Na3VO4）对椎间盘软骨终板软骨细胞的作用。方法使用酶消化法培养大鼠椎间盘软骨终板软骨细胞，以10、20及30umol／L不同浓度Na3VO4干预第三代软骨细胞。培养7d后，用MTT法检测软骨细胞增殖率，RT-PCR检测Ⅱ型、Ⅸ型胶原和蛋白聚糖（Aggrecan）mRNA的表达，定量RT—PCR检测PTPN1、IGFRmRNA的表达。结果与对照组比较：（1）10umol／L Na3VO4组上述各项检测指标变化不明显，差异无统计学意义；（2）20及30umol／L Na3VO4组软骨细胞增殖率下降（q=31．51，81．42，P〈0．01）、Aggrecan（q=52．09，102．55，P〈0．01）和PTPN1（q=7．67，4．74，P〈0．01）mRNA表达降低；（3）30umol／L Na3VO4组Ⅱ型胶原（q=51．46，P〈0．01）、Ⅸ型胶原（q=8．62，P〈0．01）mRNA表达降低；（4）各浓度Na3VO4组IGFRmRNA均增加（q=13．96，7．67，4．74，P〈0．01）。结论20及30umol／L Na3VO4可抑制椎间盘软骨细胞增殖，降低Ⅱ、Ⅸ型胶原、Aggrecan以及PTPN1的mRNA表达，说明Na3VO4对软骨细胞中PTPs的活性有抑制作用，从而降低了软骨细胞生物学功能。但Na3Vo4对IGFR mRNA无抑制作用，并可能有一定的促表达作用。Objective To evaluate the effect of Sodium Orthovanadate （Na3Vo4） on the chondrocyte in the end plate of intervertebral discs. Methods Chondrocyte cells got from the end plate of rat intervertebral disc were cultured by enzyme digestive method. Different density Na3Vo4 （10, 20 and 30 umol/L） was used to this system and then the cells were cultured for 7 days. The growth rate of chondrocyte cells were examined by MTT test, the expression of type Ⅱ collagen, type Ⅸ collagen and Aggrecan mRNA were detected by RT-PCR test. The expression of PTPNI and IGFR mRNA were determined by real time RT-PCR test. Results In the measurement performed at 7 day, compare with control group, there was no significant change of the growth rate of chondrocytes and the expression of type Ⅱ collagen, type IX collagen, Aggrecan, PTPN1 and IGFR mRNA in the 10 umol/L Na3Vo4. The growth rate （q=31.51, 81.42, P〈 0.01）, Aggrecan mRNA （q=52.09, 102.55, P〈 0.01） and PTPN1 mRNA （q=7.67, 4.74, P〈 0.01） of chondrocyte cells decreased significantly of both 20 and 30 umol/L Na3Vo4 groups. The expression of type Ⅱ collagen （q =51.46, P〈 0.01 ）, type Ⅸ collagen mRNA （q =8.62, P〈 0.01） of chondrocytes decreased significantly of 30 umol/L Na3Vo4 groups, too. The expression of IGFR mRNA （q=13.96, 7.67, 4.74, P〈 0.01） of chondrocytes increased significantly of each of Na3Vo4 groups. Conclusion 20 and 30 umol/L Na3Vo4 play important effects in the growth rate of chondrocytes and the expression of type Ⅱ collagen, type Ⅸ collagen, Aggrecan and PTPN1 mRNA. But there is not any depressant effect in the expression of IGFR mRNA. Na3Vo4 inhibits the expres- sion of PTPN1 gene in PTPs and depresses the biological function of chondrocyte. There is not any depressant effect for IGFR mRNA in each of Na3Vo4 groups, possibly with the advance of the express.

关 键 词：椎间盘 软骨细胞 钒酸盐类 

分 类 号：R686[医药卫生—骨科学]