中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2011年
41期
2912-2916
,共5页
徐宏光%彭红心%程加峰%吕坤
徐宏光%彭紅心%程加峰%呂坤
서굉광%팽홍심%정가봉%려곤
软骨细胞%蛋白聚糖类%胶原
軟骨細胞%蛋白聚糖類%膠原
연골세포%단백취당류%효원
Chondrocytes%Proteoglycans%Collagen
目的 建立人颈椎椎体终板软骨细胞退变模型,观察人正常颈椎椎体和退变颈椎椎体终板软骨细胞的形态及表征.方法 选择2010年7月至2011年7月49例颈椎骨折、脱位(19例)及颈椎病(30例)患者术中取出的颈椎终板软骨,用酶消化法分别分离培养人正常颈椎椎体终板软骨细胞(对照组)和退变颈椎椎体终板软骨细胞(颈椎病组)；用倒置显微镜和HE染色法观察细胞形态学变化；四甲基偶氮唑蓝(MTT)法绘制细胞生长曲线；甲苯蓝染色及反转录-PCR(RT-PCR)法对终板软骨细胞进行鉴定；RT-PCR法检测终板软骨细胞特征性基因蛋白多糖、Ⅱ型胶原及Ⅰ型胶原的表达.结果 人颈椎椎体终板软骨细胞表达特征性蛋白多糖、Ⅱ型胶原及Ⅰ型胶原,其生长情况及细胞表型类似于关节软骨细胞.对照组原代终板软骨细胞以多角形为主,增殖速度较快；而颈椎病组原代终板软骨细胞以梭形为主,细胞增殖速度较慢.颈椎病组原代终板软骨细胞表达的蛋白多糖基因(0.695 ±0.052)和Ⅱ型胶原基因(0.726 ±0.035)均低于对照组(0.950±0.032、0.907±0.078,t=7.263、3.681,P=0.002、0.021),Ⅰ型胶原基因则高于对照组(0.795±0.028比0.552±0.070,t=-5.560,P=0.005).结论 成功建立了人颈椎椎体终板软骨细胞退变模型,为椎间盘退变机制研究提供了较好的细胞学基础,解决了以前一直以动物细胞模型为研究对象的局限性.
目的 建立人頸椎椎體終闆軟骨細胞退變模型,觀察人正常頸椎椎體和退變頸椎椎體終闆軟骨細胞的形態及錶徵.方法 選擇2010年7月至2011年7月49例頸椎骨摺、脫位(19例)及頸椎病(30例)患者術中取齣的頸椎終闆軟骨,用酶消化法分彆分離培養人正常頸椎椎體終闆軟骨細胞(對照組)和退變頸椎椎體終闆軟骨細胞(頸椎病組)；用倒置顯微鏡和HE染色法觀察細胞形態學變化；四甲基偶氮唑藍(MTT)法繪製細胞生長麯線；甲苯藍染色及反轉錄-PCR(RT-PCR)法對終闆軟骨細胞進行鑒定；RT-PCR法檢測終闆軟骨細胞特徵性基因蛋白多糖、Ⅱ型膠原及Ⅰ型膠原的錶達.結果 人頸椎椎體終闆軟骨細胞錶達特徵性蛋白多糖、Ⅱ型膠原及Ⅰ型膠原,其生長情況及細胞錶型類似于關節軟骨細胞.對照組原代終闆軟骨細胞以多角形為主,增殖速度較快；而頸椎病組原代終闆軟骨細胞以梭形為主,細胞增殖速度較慢.頸椎病組原代終闆軟骨細胞錶達的蛋白多糖基因(0.695 ±0.052)和Ⅱ型膠原基因(0.726 ±0.035)均低于對照組(0.950±0.032、0.907±0.078,t=7.263、3.681,P=0.002、0.021),Ⅰ型膠原基因則高于對照組(0.795±0.028比0.552±0.070,t=-5.560,P=0.005).結論 成功建立瞭人頸椎椎體終闆軟骨細胞退變模型,為椎間盤退變機製研究提供瞭較好的細胞學基礎,解決瞭以前一直以動物細胞模型為研究對象的跼限性.
목적 건립인경추추체종판연골세포퇴변모형,관찰인정상경추추체화퇴변경추추체종판연골세포적형태급표정.방법 선택2010년7월지2011년7월49례경추골절、탈위(19례)급경추병(30례)환자술중취출적경추종판연골,용매소화법분별분리배양인정상경추추체종판연골세포(대조조)화퇴변경추추체종판연골세포(경추병조)；용도치현미경화HE염색법관찰세포형태학변화；사갑기우담서람(MTT)법회제세포생장곡선；갑분람염색급반전록-PCR(RT-PCR)법대종판연골세포진행감정；RT-PCR법검측종판연골세포특정성기인단백다당、Ⅱ형효원급Ⅰ형효원적표체.결과 인경추추체종판연골세포표체특정성단백다당、Ⅱ형효원급Ⅰ형효원,기생장정황급세포표형유사우관절연골세포.대조조원대종판연골세포이다각형위주,증식속도교쾌；이경추병조원대종판연골세포이사형위주,세포증식속도교만.경추병조원대종판연골세포표체적단백다당기인(0.695 ±0.052)화Ⅱ형효원기인(0.726 ±0.035)균저우대조조(0.950±0.032、0.907±0.078,t=7.263、3.681,P=0.002、0.021),Ⅰ형효원기인칙고우대조조(0.795±0.028비0.552±0.070,t=-5.560,P=0.005).결론 성공건립료인경추추체종판연골세포퇴변모형,위추간반퇴변궤제연구제공료교호적세포학기출,해결료이전일직이동물세포모형위연구대상적국한성.
Objective To establish an in vitro model of degeneration of human cervical endplate chondrocytes and observe the morphology and phenotypes of endplate chondrocytes in normal and degenerative cervical vertebral endplates.Methods Cartilage endplates of 49 patients were divided into control group ( n =19 ) with cervical vertebral fracture or dislocation and experiment group ( n =30) with cervical spondylotic myelopathy.Endplate chondrocytes were isolated by enzyme digestion and cultured in vitro.The morphological appearances,growth curve and biological characteristics of endplate chondrocytes from normal and degenerative cartilage endplate were observed by inverted phase contrast microscope,HE staining,MTT,teluidine blue staining and reverse transcription-polymerase chain reaction (RT-PCR)respectively.RT-PCR was used to detect the mRNA expression of aggrecan,type Ⅱ collagen and type Ⅰcollagen.Results The endplate chondrocytes expressed aggrecan,type Ⅱ collagen and type Ⅰ collagen.The phenotypes and biological characteristics were similar to those of articular chondrocytes.The morphological appearance of primary endplate chondrocytes in the control group were mostly polygons,nucelus with round or ellipse,sometimes nuclei,vacuolus in intracytoplasm,expressing a high proliferating rate.The cells of the experiment group were fusiform and their proliferating rates decreased.Compared with the control group,the mRNA expression of aggrecan ( 0.695 ± 0.052 vs 0.950 ± 0.032,t =7.263,P =0.002) and type Ⅱ collagen (0.726 ± 0.035,0.907 ± 0.078,t =3.681,P =0.021 ) markedly decreased.And the mRNA expression of type Ⅰ collagen (0.795 ±0.028 vs 0.552 ±0.070,t =-5.560,P =0.005 )increased in the experiment group.Conclusion A degenerative cell model of human cervical endplate chondrocytes has been established successfully in vitro.It may offer the cytological rationales for exploring the mechanism of intervertebral disc degeneration.And the previous restrictions of studying only the model of animal cells shall be resolved.
