碱性成纤维细胞生长因子对犬隐静脉间质细胞体外培养抗钙化作用  

The anti-calcification effect of basic fibroblast growth factoron canine saphenous vein interstitial cells in vitro

作　　者：宿学家[1] 刘维永[1] 陈文生[1] 卫向阳[1] 欧阳辉[1] 顾春虎[1] 

机构地区：[1]第四军医大学西京医院全军心血管病研究所心血管外科,西安710032

出　　处：《中华实验外科杂志》2006年第9期1050-1052,i0002,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(30271292)

摘　　要：目的观察作为组织工程瓣间质种子细胞的犬隐静脉间质细胞体外培养钙化机制,探讨碱性成纤维细胞生长因子(bFGF)对犬隐静脉间质细胞体外培养钙化的抑制作用。方法分离培养犬隐静脉间质细胞,分为正常培养对照、促钙化培养和bFGF培养3组。2周后行von kossa染色,检测细胞层钙沉积、碱性磷酸酶(ALP)活性及上清液中骨钙素(OC)含量,细胞ALP、OC mR- NA表达。结果培养2周后,促钙化组及对照组均有细胞结节形成,bFGF组细胞呈极性密集生长,无结节形成。von kossa染色促钙化组呈强阳性,bFGF组及对照组呈弱阳性。促钙化组钙沉积及ALP活性比对照组均明显增高(5．384±0．443比2．198±0.173,P＜0.01),ALP高2．13倍(9．611±0．639比4.819±0．334,P＜0．01),bFGF使两者均显著降低(0．192±0．024比5．384±0.443,P＜0．01,3．069±0．178比9．611±0．639,P＜0．01)。促钙化组上清液OC含量于第6天始升高,至第12天到达高峰,然后有轻度下降。bFGF组及对照组均维持较低水平。结论体外培养的犬隐静脉间质细胞在促钙化条件下具有成骨细胞的表型,有明显的钙化现象发生,bFGF具有抑制其钙化的作用。Objective To observe the anti-calcification effect of basic fibroblast growth factor （bFGF） on canine saphenous vein interstitial cells （SVICs） in vitro. Methods Canine SVICs were cultured and devided into control group, calcific group and bFGF group. After 2 weeks,cellular layer was stained by von kossa. Calcium deposition and alkaline phospatase （ALP） activity of cellular layer,osteocaltin （OC） in cultured supernatant were analyzed. Results （ 1 ） After 2 weeks, in calcific group, caleific nodules were enriched by von kossa staining. In control group, there were cellular nodules but calcification was mildly positive. In bFGF group, there was no cellular nodules and calcification was negative by von kossa staining; （2） Calcium deposition in calcific group was 27 times higher than that of control group [ （5. 384 ± 0. 443） vs （0. 192 ± 0. 024 ）, P 〈 0.01 ]. The activity of ALP in the calcific group was 2.13 times higher than of control group [ （9. 611 ± 0. 639） vs （3. 069 ± 0. 178）, P 〈 0.01 ]. Both calcium and ALP activity were very lower in bFGF group [（2.198± 0.173） vs （5.384± 0.443）, P〈 0.01; （4. 819 ± 0. 334） vs （9. 611 ± 0. 639）, P 〈 0.01 ]. OC of supernatant was increased at 6th day, reached the peak at 12th day, and then slightly decreased in calcific group. But in bFGF and control groups the content of OC was maintained at a lower level. Conclusion The cultured canine SVICs have osteoblast phenotype and are calcified in calcified medium in vitro, bFGF can inhibit the calcification significantly.

关 键 词：组织工程 瓣 种子细胞 钙化 碱性成纤维细胞生长因子 

分 类 号：R318.0[医药卫生—生物医学工程]