摘要
背景:基础研究表明Ⅰ型胶原蛋白在骨生成、骨的质量及骨折方面作用突出,同时参与骨融合过程,但关于此基因多态性与脊柱融合的关系鲜见报道。目的:观察Ⅰ型胶原蛋白基因多态性,并探讨其与金属植入物及自体骨移植脊柱融合效果的相关性。方法:随机选择青岛大学医学院附属医院住院患者中需行脊柱融合的自愿受试者200例,其中行颈椎前路椎体次全切除减压自体髂骨植骨融合钛板内固定者102例,腰椎后路椎板切除减压横突间植骨融合内固定者98例,以223名健康正常人作对照。抽取受试者外周静脉血,并提取白细胞DNA;采用聚合酶链反应扩增目的基因所在的DNA片段,扩增片段长度为293bp;应用聚合酶链反应-限制性片段长度多态性方法检测外周血白细胞基因组Ⅰ型胶原蛋白基因Pcol2位点多态性,扩增产物经限制性内切酶Eco31I酶切,琼脂糖凝胶电泳观察,有酶切位点用G基因表示,无酶切位点用T基因表示;术前及术后3,6,12个月的X射线片观察植骨融合情况,按照快(<3个月)、中(3~6个月)、慢(7~12个月)分期进行对比。结果与结论:两组受试者均存在Ⅰ型胶原蛋白基因Pcol2位点-1997G/T多态性,GG型166例,GT型232例,TT型25例,并发现GG基因型与颈椎前路减压自体髂骨植骨融合有相关性(P=0.004),该基因型在快组中占50%,明显高于中组(33.3%)和慢组(16.7%);-1997G/T多态性与腰椎后路横突间植骨融合无相关性(P=0.831)。结果提示Ⅰ型胶原蛋白基因Pcol2位点-1997G/T单核苷酸多态性GG基因型可能是促进颈椎前路自体髂骨植骨融合的重要遗传基因。
BACKGROUND:Basic research demonstrated that type I collagen exhibited prominent effect on osteogenesis,bone mass and bone fracture,which also participated in the bone fusion.However,few reports concerning the polymorphism of type I collagen gene and spinal fusion.OBJECTIVE:To investigate the polymorphism of type I collagen and to explore its relationship with the spinal fusion rate following metal implant or autogenous bone transplantation.METHODS:A total of 200 volunteers who need to receive spinal fusion in the Affiliated Hospital of Qingdao University Medical College were selected,including 102 cases received anterior cervical subcorpectomy combined with iliac bone implantation fusion following decompression,and 98 cases received posterior laminectomy for decompression combined with intertransverse process fusion.Meantime,223 normal adults were served as the control group.The peripheral blood was drawn-off and genomic DNA was extracted from white blood cells.The specific fragment which includes the objective gene was amplified by polymerase chain reaction(PCR),with length of 293 bp.The genotypes of Pcol2 site in type I collagen were detected by PCR-restriction fragment length polymorphism(PCR-RFLP) method.The PCR product was digested with restriction endonuclease Eco31I and the result was observed by agarose gel electrophoresis.The G gene represented for the presence of the restriction endonuclease site,while the T gene for the absence of the restriction endonuclease site.The fusion rate of the bone graft was evaluated by x-ray film prior to and at months 3,6 and 12 after operation,and the results were compared by stages including quick(3 months),middle(3-6 months) and slow(6-12 months).RESULT AND CONCLUSION:There were the-1997G/T polymorphisms of the type I collagen gene in 423 cases,including 166 cases with GG,232 cases with GT,and 25 cases with TT,in addition,there was some correlation between the GG genotype and the iliac bone implantation fusion(P =0.004).The GG genotype accounted for 50% in the fast group,which was obviously greater than that of the middle and slow groups(33.3% and 16.7%,respectively).However,the-1997G/T polymorphisms had no correlation with the bone graft fusions inter transverse process of lumbar vertebra(P=0.831).The GG genotype in the-1997G/T polymophsim of the type I collagen gene may be the essential factor which can promote the C-spine auto-ilium graft fusion.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2010年第9期1607-1611,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research