CD24在人髓核细胞鉴定应用中的相关研究

发布时间：2018-05-23 09:38

  本文选题：髓核细胞 + 细胞鉴定　； 参考：《昆明医科大学》2015年硕士论文

【摘要】：目的：CD24是一种低分子质量高度糖基化的粘附分子,表达于人体多种细胞表面。该研究我们通过对比人髓核与软骨中CD24的表达,探讨CD24在鉴别髓核细胞中的作用及效果。方法：通过收集手术治疗中患者的椎间盘及软骨组织,患者年龄5～71岁,共27例,手术原因包括先天性脊柱侧凸、外伤、椎间盘退变。其中标本组织切片HE染色观察组织,另一部分标本的髓核组织和软骨组织提取RNA后逆转录为cDNA进行OD值测定,普通凝胶电泳PCR、RT-PCR测定组织中CD24的表达情况并进行对比。结果：1.大体观察：健康髓核外观呈乳白或灰白色半透明或不透明的胶冻样物质,富有弹性.退变髓核组织分界清晰,髓核经髓核钳咬出后呈絮状灰白或淡黄色组织,其组织弹性稍差2.组织切片HE染色：髓核组织结构清晰,纤维环染色均匀,髓核细胞呈巢状排列,无退变患者髓核细胞数量丰富,细胞形态规整,细胞突长度适中,轻度退变髓核组织内细胞数量较正常髓核内轻度减少,细胞形态尚规整,退变组织纤维环内偶见少量巨噬细胞。软骨组织染色均匀,可见软骨陷凹,软骨细胞染色较深未见其他组织结构。3.RNA紫外分光度仪器检测结果：本次实验中所提取的RNA经检测OD值与浓度,提示RNA纯度较高,浓度满足后续实验要求。4.普通凝胶电泳荧光PCR结果：可见髓核与软骨内参基因产物条带位置大约为200dp左右,条带清晰,见髓核CD24通过PCR扩增后产物有清晰条带位置大概为150dp,而软骨CD24特异性扩增处未见明显条带。5.RT-PCR结果：实验各组髓核内CD24表达与软骨内表达差异均有显著统计学意义(PO.01),各组髓核内CD24的表达差异没有统计学意义(PO.05),髓核内CD24的表达与年龄之间没有明显相关关系(相关系数rs=0.124,P0.05)结论：1. Pfirrmann Ⅰ～Ⅲ级髓核组织主要细胞成分为髓核细胞；2.CD24在髓核组织与软骨组织之间表达存在明确的统计学差异,CD24可以作为Ⅱ型胶原蛋白和蛋白聚糖之后的特异性鉴定因子；3.不同年龄、不同疾病类型、Pfirrmann Ⅰ～Ⅲ型的髓核组织内CD24的表达没有统计学意义,CD24与髓核细胞的功能及退变程度没有明显相关性；
[Abstract]:Objective: to express the low molecular weight and high glycosylated adhesion molecule on the surface of many human cells. In this study, we compared the expression of CD24 in nucleus pulposus and cartilage to explore the role and effect of CD24 in differentiating nucleus pulposus cells. Methods: 27 patients (571 years old) with congenital scoliosis, trauma and disc degeneration were collected. The tissue sections were stained with HE, and the other part of the tissue of nucleus pulposus and cartilage were extracted from the nucleus pulposus and chondrocytes. The OD value was measured by reverse transcription to cDNA. The expression of CD24 in the tissue was detected by RT-PCR and compared. The result is 1: 1. General observation: the appearance of healthy nucleus pulposus is milky white or grayish-white translucent or opaque gelatinous material, elastic. The tissue boundary of degenerative nucleus pulposus was clear. The nucleus pulposus showed flocculent grayish white or light yellow tissue after bite of nucleus pulposus forceps, and its tissue elasticity was slightly less than 2. HE staining showed clear structure of nucleus pulposus, uniform staining of fibrous ring, nesting arrangement of nucleus pulposus cells, abundant number of nucleus pulposus cells, regular morphology of cells and moderate length of cell processes in patients without degeneration. The number of cells in the mild degenerative nucleus pulposus was slightly lower than that in the normal nucleus pulposus, and the morphology of the cells was still regular, and a few macrophages were occasionally found in the fibrous ring of the degenerative tissue. Cartilage tissue staining was uniform, cartilage concave was visible, and other tissue structures were not found in chondrocyte staining. 3. The RNA extracted in this experiment was detected by OD value and concentration, indicating that the purity of RNA was high. The concentration meets the requirements of subsequent experiments. The results of fluorescence PCR showed that the position of gene products in nucleus pulposus and cartilage was about 200dp and the bands were clear. It was found that the clear band position of CD24 amplified by PCR was about 150dp, but there were no obvious bands in the specific amplification of cartilage CD24. 5. RT-PCR results: there were significant differences between the expression of CD24 in nucleus pulposus and the expression in cartilage in each group. 5. The results of RT-PCR showed that there were significant differences in the expression of CD24 in the nucleus pulposus and in the cartilage. There was no significant difference in the expression of CD24 in nucleus pulposus (P < 0.05). There was no significant correlation between the expression of CD24 in nucleus pulposus and age (r = 0.124, P 0.05). Conclusion: 1. The main cell component of Pfirrmann 鈪，

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