精神分裂症外周血P细胞表面抗原分型及其超微结构实验研究

发布时间：2018-02-01 07:41

本文关键词： 精神分裂症 P细胞细胞亚型超微结构表面抗原　出处：《遵义医学院》2017年硕士论文　论文类型：学位论文

【摘要】：目的:研究精神分裂症患者外周血P细胞表面抗原分型及其超微结构改变,进而推测P细胞在精神分裂症发病和诊断中可能存在的作用。方法:采集50例外周血标本分两组,其中25例精神分裂症(男12,女13,年龄:32.84±2.21),25例正常对照(男18,女7,年龄:30.40±1.76)。将外周血标本涂片,经瑞氏染色后进行显微镜下淋巴细胞分类计数。同时用淋巴细胞提取液Percoll TM提取外周血单个核细胞,之后用免疫磁珠法将单个核细胞按照细胞表面抗原分离纯化为CD4+、CD8+、B淋巴细胞三个亚型。分离的亚型细胞分为两部分,一部分经涂片瑞氏染色后,用于鉴定P细胞亚型。另一部分亚型细胞沉淀后用醋酸双氯铀-柠檬酸铅负染,采用透射电子显微镜检查超微结构。用流式细胞术检测两组外周血淋巴细胞表面抗原(CD3+,CD4+,CD8+,CD16+,CD56+,CD19+)。结果:精神分裂症外周血P细胞百分比为34.53±9.92比正常对照的9.79±3.45升高,差异有显著性(P0.05)。精神分裂症外周血中分离得到的CD4+、CD8+、B淋巴细胞亚型的涂片中均能观察到P细胞,主要表现为胞质嗜碱,细胞核大,核质比增大,核型不规则,染色质纤细,常见一到两个核仁。P细胞电镜结果显示,细胞增大,胞质量少,细胞核大,核质比增加。核形异常,核仁明显,常染色质丰富,异染色质凝集呈块。胞质内可见线粒体聚集分布,峭丰富。线粒体常呈现轻度扩张,变性肿胀,甚至空化的形态改变。细胞表面微绒毛丰富。正常淋巴细胞电镜下表现为,细胞大小适中,胞质丰富,细胞核常呈圆形或椭圆形,异染色质较常染色质丰富,分布在核膜周围或细胞核中间。细胞表面可有微绒毛。流式细胞仪检测两组外周血淋巴细胞表面标记物,其中精神分裂症CD4+百分比为38.64±1.68比正常对照的35.17±1.26升高,精神分裂症CD4+绝对计数为1019.16±130.75比正常对照的667.86±49.88升高,精神分裂症CD19+百分比为14.06±1.06比正常对照的10.42±0.60升高,精神分裂症CD19+绝对计数为428.06±107.22比正常对照209.02±31.04升高,而精神分裂症CD16+CD56+百分比为11.81±1.22比正常对照16.19±1.60降低,所有差异均有显著性(P0.05)。结论:精神分裂症外周血P细胞百分比显著增加。按照表面抗原分为的CD4+、CD8+、B淋巴细胞三个亚型细胞中均含有P细胞。P细胞与正常淋巴细胞相比超微结构有差异,主要表现为细胞增大,核质比增大,常染色质丰富,异常核形,常可见1到2个核仁,线粒体形态异常,主要表现为线粒体峭扩张。精神分裂症组外周血细胞表面抗原CD4+、CD19+百分比和绝对计数升高,CD16+CD56+百分比降低。
[Abstract]:Objective: to study the surface antigen typing and ultrastructural changes of P cells in patients with schizophrenia. Methods: 50 cases of peripheral blood samples were collected and divided into two groups, 25 cases of schizophrenia (male 12, female 13). 25 normal controls (male 18, female 7, age 30. 40 卤1. 76). The peripheral blood samples were smeared with age: 32. 84 卤2. 21 and 25 normal controls (male 18, female 7, age 30. 40 卤1. 76). The peripheral blood mononuclear cells (PBMC) were extracted by Percoll TM, the lymphocyte extract was used to extract the peripheral blood mononuclear cells (PBMC). Then the mononuclear cells were purified into three subtypes of CD4 CD8 B lymphocytes by immunomagnetic beads according to the cell surface antigen. The isolated subtypes were divided into two parts. Some of them were used to identify the P cell subtype after smear staining, and the other one was stained negatively with dichlorouranium-citrate lead acetate after precipitation. The ultrastructure was examined by transmission electron microscope and CD16 CD56 was detected by flow cytometry. Results: the percentage of P cells in peripheral blood of schizophrenia was 34.53 卤9.92, which was higher than that of normal controls (9.79 卤3.45). The difference was significant (P 0.05). P cells were observed in the smears of CD4 CD8 B lymphocyte subtypes isolated from the peripheral blood of schizophrenia, mainly as cytoplasmic basophils. The nucleus was large, the ratio of nucleus to cytoplasm was increased, the karyotype was irregular, the chromatin was slender. The electron microscopic results of one or two nucleolus. P cells showed that the cells were enlarged, the cell mass was small, the nucleus was large, the ratio of nucleus to cytoplasm was increased, and the nucleus shape was abnormal. The nucleoli are obvious, euchromatin is rich, heterochromatin agglutination is a mass. Mitochondria can be seen in the cytoplasm. Mitochondria are often slightly dilated and swollen. Even the morphologic changes of cavitation. The microvilli on the surface of the cells were abundant. The normal lymphocytes showed moderate size, abundant cytoplasm, round or elliptical nuclei and more heterochromatin than normal chromatin. The microvilli could be found on the surface of the cells. Flow cytometry was used to detect the surface markers of peripheral blood lymphocytes in the two groups. The percentage of CD4 in schizophrenia was 38.64 卤1.68, which was higher than that in normal controls (35.17 卤1.26). The absolute CD4 count of schizophrenia was 1019.16 卤130.75 and 667.86 卤49.88 in normal controls. The percentage of CD19 in schizophrenia was 14.06 卤1.06 compared with 10.42 卤0.60 in normal controls. The absolute CD19 counts of schizophrenia were 428.06 卤107.22 and 209.02 卤31.04 respectively. The percentage of CD16 CD56 in schizophrenia was 11.81 卤1.22 and 16.19 卤1.60 in normal controls. Conclusion: the percentage of P cells in peripheral blood of schizophrenia was significantly increased, and CD4 CD8 was classified according to surface antigen. The ultrastructure of P cells in the three subtypes of B lymphocytes was different from that of normal lymphocytes. The main features were the enlargement of cells, the increase of nuclear / cytoplasm ratio, the abundance of normal chromatin and the abnormal nucleation of B lymphocytes. One or two nucleoli were often found with abnormal mitochondrial morphology, mainly characterized by mitochondrial dilatation. In schizophrenia group, the percentage and absolute count of peripheral blood cell surface antigen (CD4) CD19 were increased. The percentage of CD16 CD56 decreased.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.3;R446.1

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