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1、不同应力条件对体外培养成肌细胞命运影［摘要］目的：探讨对体外培养成肌细胞施加不同牵张应力后细胞命运的改变，为临床合理施加矫形力提供理论依据。方法：利用Flexercell细胞加力仪器与弹性加力板对体外培养C2C12成肌细胞施加不同频率、幅度的牵张应力，形变量代表应力幅度。通过MTT实验和PARP剪切实验测定在受到不同大小的牵张应力作用下(0%,5%,10%,15%,20%/10cycles/min)的C2C12细胞的增殖和凋亡情况，通过检测细胞分化相关基因的表达改变，分析应力条件下细胞分化状态的改变。结果：5%的周期性应力可以促进细胞的增殖
2、，同时抑制分化培养基诱导的细胞分化相关基因myogenin等的表达。当牵张应力大于10%后，可以观察到明显的DNA断裂。除此以外，我们还发现牵张应力大于10%后，细胞内开始出现大量的PARP剪切产物，随应力的增加而增多。当周期性牵张应力超过15%10cycles/min时，整体存活细胞数明显降低。结论：高强度牵拉介导的细胞凋亡，参与整体细胞数的减少，可能不利于组织的重塑和再生。［关键词］周期性牵张应力；凋亡；增殖；分化［中图分类号1Q813.1［文献标识码］A［文章编号］1008-6455(2011)07-1087-03Correlatio
3、nbetweencyclicstretchmagnitudesandmyoblastscellfateinvitroKUANGWeil,TANJia~li2,ZHANGHong-mei3,DUANJian-minl,WANGWei-jianl,LIXiaol(1.DepatmentofStomatology,GuangzhouGeneralHospitalofGuangzhouMilitaryCommand,Guangzhou510010,Guangdong,China；2.DepartmentofOrthodontics,Schoolof
4、Stomatology,TheFourthM订itaryMedicalUniversity；3.DepartmentofStomatology,TangduHospital,TheFourthMilitaryMedicalUniversity)Abstract：ObjectiveToinvestigatethedetailedeffectsofthecyclicstretchesofaserialofmagnitudesoncellgrowth,differentiationandapoptosis・C2C12myoblastsweresu
5、bjectedto0%,5%,10%,15%,20%/10cycles/mincyclicstretch・MethodsFlexercellstretchunitwasappliedinthisstudy.Wefirstevaluatedcellviabilityuponserialcyclicstretches(0%,5%,10%,15%,20%/10cycles/min).MTTassaywasappliedtotestcellviabilityandproliferatiori.WesternblotofcleavedPARPwasd
6、onetotesttheapoptosis・Inaddition,cellwereinducedinmyogenicdifferentiationmediumunderserialcyclicstretches,differentiationgenemarkermyogeninwastestedbyRT-PCR・ResuItsWhentheextensionmagnitudeisabout5%,cellgrowthwasfacilitated・Incontrast,myogenicmediuminducedcelldifferentiati
7、onunderthisconditionwasinhibitedasseenbymyogeninexpression・Withtheextensionmagnitudesexceeding15%lOcycles/min,cellviabilitydecreased.SincebothproliferationandcelldeathaffectedtheMTTresults,wethentestedwhetherapoptosiswasinvolvedintheprocess・AbundantPARPcleavagewasobservedw
8、henthecyclicstretcheswereover15%・ConclusionsLowmagnitudeofstretchinhibitedproliferationan
