血小板衍生生长因子-BB对破骨细胞功能影响的实验研究

The effects of platelet-derived growth factor-BB on the osteoclastic bone resorption

目的探讨血小板衍生生长因子 -BB(platelet-derivedgrowthfactor-BB,PDGF -BB)对破骨细胞生物学功能的影响。方法 (1)利用酶消化法分离培养成人破骨细胞 ;(2)应用透射电子显微镜方法观察破骨细胞对PDGF -β 受体的表达 ;(3)在经过纯化的破骨细胞上清液中施加重组人基因PDGF -BB ,利用酶动力学方法测定培养上清液中的酸性磷酸酶 (ACP)和抗酒石酸酸性磷酸酶(TRAP)的活性 ;(4)通过Leica图像分析仪观察在PDGF -BB作用下 ,破骨细胞所形成骨吸收陷窝的数目与面积。结果 (1)破骨细胞膜上有胶体金颗粒沉着 ;(2)破骨细胞培养上清液中的ACP和TRAP活性随着PDGF -BB浓度的升高而增加 ,分别从(1.85±0.13)u/L和(1.73±0.15)u/L(对照组 )增加至(2.86±0.15)u/L和(2.75±0.24)u/L(40μg/L组 ,P<0.01) ;(3)在PDGF -BB的作用下 ,破骨细胞所形成的骨吸收陷窝的面积从(435.08±237.50)μm2(对照组 )增高至(630.26±240.64)μm2 (40μg/L组 ,P<0.01) ,每个骨片上骨吸收陷窝的数目亦从14.00±1.41增加为26.00±2.00(P<0.05或P<0.01)。结论PDGF -BB通过与PDGF -β受体结合 。

ObjectiveTostudytheeffectofplatelet-derivedgrowthfactor-BB(PDGF -BB)onthe biologicalfunctionsofhumanosteoclasts.Methods1)Adultosteoclastswereisolatedfromhumanspongy bonewithcollagenaseⅡ.2)PDGFreceptor- βwasdetectedontheosteoclastswithtransmissionelectronmi croscope.3)ThePDGF -BBwasexertedontheosteoclaststhatadheredtothebonesliceattheconcentration of0,10,20,30and40μg/L.Theacidphosphatase(ACP)andthetartrate -resistantacidphosphatase (TRAP) activitiesintheculturemediumweremeasuredwithkineticmethod.4)Thenumberandareaoflacunaeformed byosteoclastsabsorptionundertheactionofPDGF -BBwereanalysedwithLeicaimageanalysisapparatus. Results1)Thecolloidalgolddepositedonthecellularmembrane.2)TheACPandTRAPactivitiesrosewith thedoseofPDGF -BB,increasingrespectivelyfrom(1.85±0.13)u/Land(1.73±0.15)u/L(control)to (2.86±0.15)u/Land(2.75±0.24)u/LwhenPDGF -BBwasat40μg/L(P<0.01).3)Thevolumeof Howship slacunaeaugmentedfrom(435.08±237.50)μm2(control)to(630.26±240.64)μm2(P<0.01), andthenumberoftheresorptionlacunaincreasedalsofrom14.00±1.41to26.00±2.00(P<0.01). ConclusionThehumanosteoclasthasPDGFreceptor- βonitsmembraneandPDGF -BBcouldstimulate osteoclasticboneresorptiondirectlyafterbindingtothereceptor - β.