CAJ | 학술논문

背景：椎间盘退变导致椎间隙狭窄的主要变化是髓核细胞表达软骨特异性基因产物蛋白聚糖和Ⅱ型胶原的减少。 　　目的：对成人的正常与退变椎间盘髓核细胞Ⅱ型胶原蛋白免疫荧光染色及番红O染色进行定量观察比较。 　　方法：取成人脊柱侧弯患者与椎间盘突出症患者自愿者术中取出的废弃髓核组织各3例，经培养后每个患者各测量26个细胞，正常组和退变组分别测量78个细胞。对正常与退变椎间盘髓核细胞进行番红“O”染色并行灰度及测定细胞内Ⅱ型胶原蛋白行免疫荧光染色定量测定。 　　结果与结论：免疫荧光染色显示：退变椎间盘髓核细胞仅轻度染色，染色模糊，其呈类圆形，纺锤形，梭形及不规则形，其内仅有极少量荧光颗粒；Ⅱ型胶原表达较正常髓核细胞降低显著(P<0.05)。番红O染色显示：退变的髓核细胞肿胀，细胞核肿胀染色略淡，细胞突起延长，胞浆染色不匀伴有空泡，部分细胞崩解，细胞分布零散，混乱，可见成片坏死区。与正常髓核细胞图像灰度值相比差异无显著性意义(P>0.05)。结果表明退变的椎间盘髓核细胞减少、部分凋亡，其细胞内Ⅱ型胶原蛋白含量较正常髓核细胞显著减少。
배경：추간반퇴변도치추간극협착적주요변화시수핵세포표체연골특이성기인산물단백취당화Ⅱ형효원적감소。 　　목적：대성인적정상여퇴변추간반수핵세포Ⅱ형효원단백면역형광염색급번홍O염색진행정량관찰비교。 　　방법：취성인척주측만환자여추간반돌출증환자자원자술중취출적폐기수핵조직각3례，경배양후매개환자각측량26개세포，정상조화퇴변조분별측량78개세포。대정상여퇴변추간반수핵세포진행번홍“O”염색병행회도급측정세포내Ⅱ형효원단백행면역형광염색정량측정。 　　결과여결론：면역형광염색현시：퇴변추간반수핵세포부경도염색，염색모호，기정류원형，방추형，사형급불규칙형，기내부유겁소량형광과립；Ⅱ형효원표체교정상수핵세포강저현저(P<0.05)。번홍O염색현시：퇴변적수핵세포종창，세포핵종창염색략담，세포돌기연장，포장염색불균반유공포，부분세포붕해，세포분포령산，혼란，가견성편배사구。여정상수핵세포도상회도치상비차이무현저성의의(P>0.05)。결과표명퇴변적추간반수핵세포감소、부분조망，기세포내Ⅱ형효원단백함량교정상수핵세포현저감소。
BACKGROUND:The narrowing of intervertebral space induced by the intervertebral disc degeneration is mainly characterized by the expression of proteoglycan in nucleus pulposus cells and the reduction of col agen type II. OBJECTIVE:To quantitatively observe col agen type II protein in adult normal and degenerative intervertebral disc nucleus pulposus cells by immunofluorescence staining and safranin O staining. METHODS:The nucleus pulposus specimens were col ected from adult scoliosis patients and patients with intervertebral disc protrusion, who were al volunteers. After culture, 26 cells in each patient were measured. There were 78 cells in both normal group and degeneration group. The normal and degenerative intervertebral disc nucleus pulposus cells were subjected to safranin“O”staining, and gray values were determined;intracellular col agen type II was detected by immunofluorescence staining. RESULTS AND CONCLUSION:Immunofluorescence staining revealed that, degenerative intervertebral disc nucleus pulposus cells were only mildly stained, with the fuzzy staining, the shape was round, spindle, fusiform and irregular. There were a very smal amount of fluorescent particles within cells. The expression of col agen type II was decreased significantly compared with normal cells (P<0.05). Safranin O staining showed that, degenerative nucleus pulposus cells began to swel , the nuclei swel ed and were stained slightly, cellprocesses were prolonged, cytoplasmic dyeing was uneven accompanying with vacuole, celldisruption, scattered and chaotic distribution were visible, patches of necrosis were observed. The image gray value showed no significant difference compared with normal nucleus pulposus cells (P>0.05). The degenerative intervertebral disc nucleus pulposus cells have a smal quantity and partial y become apoptotic, the content of col egen type II protein is decreased significantly compared with normal nucleus pulposus cells.