CAJ | 학술논문

目的对比分析大骨节病儿童与成人病例外周血中的差异表达基因,为确定大骨节病软骨损伤相关基因提供科学依据.方法收集18例大骨节病儿童(儿童病例组)、18例健康儿童(儿童对照组)的外周血样,按性别与年龄匹配后分为6组.收集21例大骨节病成人(成人病例组)、21例健康成人(成人对照组)的外周血样,按性别与年龄匹配后分为7组.每组均包含3名健康人与3名病例.提取血液总RNA,利用基因芯片技术检测4组儿童(24例)与5组成人(30例)的367个外周血基因差异表达,通过平均基因表达率(表达率≥2.0或≤0.5)分别确定大骨节病儿童组与大骨节病成人组的差异表达基因.并用实时荧光定量PCR (qRT-PCR)技术检测余下的2组儿童(12例)与2组成人(12例)血样中3个基因表达,验证芯片结果的可靠性.结果儿童病例组外周血检测到101个差异表达基因,占所测基因(367个)的27.5％,少于大骨节病成人病例组(159个,43.3％,x2=20.036,P＜ 0.01);儿童病例组与成人病例组共存57个差异表达基因,其中21个基因差异表达的方向不同.qRT-PCR验证结果与基因表达芯片结果一致.结论大骨节病儿童与成人外周血基因表达模式随关节软骨和运动功能损害的严重程度不同而不同.在儿童大骨节病发病中骨软骨发育相关基因的异常表达可能与儿童骺板软骨的损伤有关,其在大骨节病早期损害中的作用值得重视.
목적 대비분석대골절병인동여성인병예외주혈중적차이표체기인,위학정대골절병연골손상상관기인제공과학의거.방법 수집18례대골절병인동(인동병례조)、18례건강인동(인동대조조)적외주혈양,안성별여년령필배후분위6조.수집21례대골절병성인(성인병례조)、21례건강성인(성인대조조)적외주혈양,안성별여년령필배후분위7조.매조균포함3명건강인여3명병례.제취혈액총RNA,이용기인심편기술검측4조인동(24례)여5조성인(30례)적367개외주혈기인차이표체,통과평균기인표체솔(표체솔≥2.0혹≤0.5)분별학정대골절병인동조여대골절병성인조적차이표체기인.병용실시형광정량PCR (qRT-PCR)기술검측여하적2조인동(12례)여2조성인(12례)혈양중3개기인표체,험증심편결과적가고성.결과 인동병례조외주혈검측도101개차이표체기인,점소측기인(367개)적27.5％,소우대골절병성인병례조(159개,43.3％,x2=20.036,P＜ 0.01);인동병례조여성인병례조공존57개차이표체기인,기중21개기인차이표체적방향불동.qRT-PCR험증결과여기인표체심편결과일치.결론 대골절병인동여성인외주혈기인표체모식수관절연골화운동공능손해적엄중정도불동이불동.재인동대골절병발병중골연골발육상관기인적이상표체가능여인동후판연골적손상유관,기재대골절병조기손해중적작용치득중시.
Objective To investigate and compare the differentially expressed genes of children and adults patients with Kashin-Beck disease (KBD) in peripheral blood mononuclear cells (PBMCs) so as to provide new clues to identify the genes related to cartilage lesion of KBD.Methods Eighteen KBD children and 18 healthy children were selected and divided into 6 matched groups according to age and gender.Twenty-one KBD adults and 21 healthy adults were selected and divided into 7 matched groups according to age and gender.Every group had 3 patients and 3 healthy persons.Peripheral blood samples were collected and total RNA was extracted respectively.The expression levels of 367 target genes in 4 child groups (24 subjects)and 5 adult groups (30 subjects) were evaluated using customized oligonucleotide microarray.Those genes with ≥2.0 or ≤0.5 expression ratio were regarded as differentially expressed genes.Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) was conducted to validate the microarray data using the remaining 2 child groups (12 subjects) and 2 adult groups (12 subjects).Results There detected 101 differentially expressed genes in KBD children,accounting for 27.5％ of all 367 target genes.It was less than that in KBD adults (159,43.3％,x2 =20.036,P ＜ 0.01).Fifty-seven differentially expressed genes were shared by KBD children and KBD adults but 21 of them were asynchronously expressed in KBD children compared to KBD adults.The result of qRT-PCR experiment was consistent with that of the microarray study.Conclusions There are significant differences in gene expression pattern between KBD children and KBD adults relating to the severity of articular cartilage and motor function damages.Bone-development related genes may play an important role in epiphyseal plate lesion of KBD children and we should pay more attention to its role in the early stage of KBD.