IL-17及其受体在人脑胶质瘤中表达的研究

发布时间：2018-09-19 14:40

【摘要】：目的：检测白细胞介素17（Interleukin-17,IL-17）及白细胞介素17受体（Interleukin-17receptor,IL-17R）在不同级别胶质瘤组织及患者外周血中表达的情况，并探讨其与胶质瘤病理分级的关系。方法：(1)病例分组：收集病理确诊的31例胶质瘤组织I级2例、II级10例、Ⅲ级9例和Ⅳ级10例为实验组，并将WHOI、Ⅱ级合并为低级别组（12例），WHOⅢ、Ⅳ级高级别组（19例）；以颅脑外伤减压术切除的失活脑组织10例为对照组；同时，31例胶质瘤患者及10例颅脑外伤患者采集静脉血标本备用，并以10例健康志愿者外周血为对照组。(2)采用免疫组织化学方法对高级别胶质瘤、低级别胶质瘤组织及其对照组中IL-17及IL-17R进行染色并检测其表达情况，研究其与胶质瘤病理分级的关系。(3)采用实时荧光定量PCR（Real-time PCR）的方法检测IL-17mRNA及IL-17RmRNA在不同级别胶质瘤组织、患者外周血及其对照组中的表达情况，研究其与胶质瘤病理分级的关系。(4)采用蛋白质免疫印记法（Western blot）检测IL-17及IL-17R在不同级别胶质瘤组织、患者外周血及其对照组中蛋白质水平的表达情况，研究其与胶质瘤病理分级的关系。结果：(1)免疫组织化学方法检测结果，IL-17在减压脑组织、低级别及高级别胶质瘤中均有阳性表达，其阳性率分别为30%、67%、100%，平均秩次分别为16.84、31.31、45.43，经各组间两两比较进行秩和检验，其表达差异均有统计学意义（HC=24.63，P＜0.05）；IL-17R在减压脑组织、低级别及高级别胶质瘤中存在阳性表达，其阳性率分别为50%、83%、100%，平均秩次分别为18.42、35.73、49.65，经各组间两两比较进行秩和检验，其表达差异均有统计学意义（HC=28.42，P＜0.05）。(2) RT-PCR方法检测结果，IL-17mRNA在对照组减压脑组织、低级别及高级别组胶质瘤组织中表达的灰度值分别为31.62±1.95、85.79±4.58、134.77±6.32，， IL-17RmRNA表达为76.20±3.97、137.66±64、193.82±6.20，经方差分析(F=221.04，p0.05)，且组间比较均有统计学差异（p0.05)。IL-17mRNA在对照组健康患者外周血、外伤减压患者外周血、低级别及高级别组胶质瘤患者外周血中表达的灰度值分别为42.09±1.65、48.21±1.79、103.71±4.37、184.67±6.51，IL-17RmRNA表达为83.18±4.57、89.47±3.82、158.45±5.34、237.32±7.29，经方差分析(F=274.64，p0.05)，对照组与减压组比较(t=-0.92，p0.05)，差异无统计学意义，余组间比较均有统计学差异（p0.05)。(3)Western blot方法检测结果，IL-17mRNA在对照组减压脑组织、低级别及高级别组胶质瘤组织中表达的灰度值分别为0.23±0.01、0.42±0.01、0.67±0.02， IL-17RmRNA表达为0.34±0.01、0.68±0.02、0.89±0.03，经方差分析(F=89.23，p0.05)，且组间比较均有统计学差异（p0.05)。IL-17蛋白质在对照组健康患者外周血、外伤减压患者外周血、低级别及高级别组胶质瘤患者外周血中表达的灰度值分别为0.33±0.01、0.35±0.01、0.54±0.02、0.85±0.02，IL-17R蛋白质的表达为0.42±0.01、0.43±0.01、0.78±0.02、0.94±0.03，经方差分析(F=107.89，p0.05)，对照组与减压组比较(t=-0.83，p0.05)，差异无统计学意义，余组间比较均有统计学差异（p0.05)。(4) IL-17在相同胶质瘤患者外周血及相对应的胶质瘤组织中的表达明显正相关(r=0.520，p=0.003)，IL-17R在相同胶质瘤患者外周血及相对应的胶质瘤组织中的表达呈显著正相关(r=0.636，p0.001)。结论：IL-17和IL-17R在不同级别胶质瘤组织和外周血中的表达有差异，且与胶质瘤的病理分级密切相关，随着胶质瘤级别的增高而增多、增强；相同胶质瘤患者外周血及相对应的胶质瘤组织中的表达有明显正相关；IL-17和IL-17R的表达可能是判断胶质瘤恶性程度的参考指标。
[Abstract]:Objective: To detect the expression of interleukin-17 (IL-17) and interleukin-17 receptor (IL-17R) in different grade glioma tissues and peripheral blood of patients, and to explore the relationship between IL-17 and pathological grading of glioma. 10 cases of grade II, 9 cases of grade III and 10 cases of grade IV were selected as the experimental group, and WHO, grade II were combined into low-grade group (12 cases), WHO, grade III and grade IV high-grade group (19 cases); 10 cases of inactivated brain tissue after craniocerebral trauma decompression were taken as the control group; meanwhile, 31 cases of glioma and 10 cases of craniocerebral trauma were taken venous blood samples for reserve, and 10 cases were healthy. The peripheral blood of the volunteers was taken as control group. (2) The expression of IL-17 and IL-17R in high-grade glioma, low-grade glioma and control group were detected by immunohistochemical staining, and the relationship between IL-17 mRNA and pathological grading of glioma was studied. (3) Real-time PCR was used to detect IL-17 mRNA and IL-17. The expression of RmRNA in different grades of glioma tissues, peripheral blood of patients and control group was studied to study the relationship between RmRNA and pathological grading of glioma. (4) The expression of IL-17 and IL-17R in different grades of glioma tissues, peripheral blood of patients and control group was detected by Western blot. Results: (1) Immunohistochemical staining showed that IL-17 was positive in decompressed brain tissues, low-grade and high-grade gliomas. The positive rates were 30%, 67%, 100%, and the average rank was 16.84, 31.31, 45.43, respectively. The difference of IL-17 expression was uniform by rank sum test. The positive rate of IL-17R was 50%, 83%, 100%, and the average rank was 18.42, 35.73, 49.65, respectively. The difference of IL-17R expression was statistically significant (HC = 28.42, P < 0.05) by rank sum test. The results showed that the gray values of IL-17 mRNA in the decompressed brain tissues, low-grade and high-grade glioma tissues were 31.62 [1.95], 85.79 [4.58], 134.77 [6.32], 76.20 [3.97], 137.66 [64], 193.82 [6.20], respectively, and the differences between the two groups were statistically significant (p0.05). The gray values of IL-17R mRNA expression in peripheral blood of healthy controls, traumatic decompression patients, low-grade and high-grade glioma patients were 42.09 [1.65], 48.21 [1.79], 103.71 [4.37], 184.67 [6.51], 83.18 [4.57], 89.47 [3.82], 158.45 [5.34], 237.32 [7.29], respectively. There was no significant difference between the two groups (t = - 0.92, p0.05). (3) The expression of IL-17 mRNA in the decompressed brain tissues, low-grade and high-grade glioma tissues was 0.23 (-0.01), 0.42 (-0.01), 0.67 (-0.02), and the expression of IL-17R mRNA was 0.23 (-0.05) by Western blot. The gray values of IL-17 protein in peripheral blood of healthy control group, traumatic decompression patients, low-grade and high-grade glioma patients were 0.33+0.01, 0.35+0.01, 0.54+0.02, 0.85 respectively. After analysis of variance (F = 107.89, p0.05), there was no significant difference between the control group and the decompression group (t = - 0.83, p0.05). There was no significant difference in the expression of IL-17R protein between the other groups (p0.05). (4) IL-17 in the peripheral blood and corresponding glioma tissues of the same glioma patients. The expression of IL-17R in peripheral blood and corresponding glioma tissues of the same glioma patients was significantly positively correlated (r = 0.520, P = 0.003). Conclusion: The expression of IL-17 and IL-17R in different grade glioma tissues and peripheral blood were significantly different, and correlated with the pathological grade of glioma. The expression of IL-17 and IL-17R in peripheral blood and corresponding glioma tissues of the same glioma patients was positively correlated. The expression of IL-17 and IL-17R may be a reference index to judge the malignant degree of glioma.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.41

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