【摘要】：目的：檢測(cè)白細(xì)胞介素17（Interleukin-17,IL-17）及白細(xì)胞介素17受體（Interleukin-17receptor,IL-17R）在不同級(jí)別膠質(zhì)瘤組織及患者外周血中表達(dá)的情況，并探討其與膠質(zhì)瘤病理分級(jí)的關(guān)系。方法：(1)病例分組：收集病理確診的31例膠質(zhì)瘤組織I級(jí)2例、II級(jí)10例、Ⅲ級(jí)9例和Ⅳ級(jí)10例為實(shí)驗(yàn)組，并將WHOI、Ⅱ級(jí)合并為低級(jí)別組（12例），WHOⅢ、Ⅳ級(jí)高級(jí)別組（19例）；以顱腦外傷減壓術(shù)切除的失活腦組織10例為對(duì)照組；同時(shí)，31例膠質(zhì)瘤患者及10例顱腦外傷患者采集靜脈血標(biāo)本備用，并以10例健康志愿者外周血為對(duì)照組。(2)采用免疫組織化學(xué)方法對(duì)高級(jí)別膠質(zhì)瘤、低級(jí)別膠質(zhì)瘤組織及其對(duì)照組中IL-17及IL-17R進(jìn)行染色并檢測(cè)其表達(dá)情況，研究其與膠質(zhì)瘤病理分級(jí)的關(guān)系。(3)采用實(shí)時(shí)熒光定量PCR（Real-time PCR）的方法檢測(cè)IL-17mRNA及IL-17RmRNA在不同級(jí)別膠質(zhì)瘤組織、患者外周血及其對(duì)照組中的表達(dá)情況，研究其與膠質(zhì)瘤病理分級(jí)的關(guān)系。(4)采用蛋白質(zhì)免疫印記法（Western blot）檢測(cè)IL-17及IL-17R在不同級(jí)別膠質(zhì)瘤組織、患者外周血及其對(duì)照組中蛋白質(zhì)水平的表達(dá)情況，研究其與膠質(zhì)瘤病理分級(jí)的關(guān)系。結(jié)果：(1)免疫組織化學(xué)方法檢測(cè)結(jié)果，IL-17在減壓腦組織、低級(jí)別及高級(jí)別膠質(zhì)瘤中均有陽性表達(dá)，其陽性率分別為30%、67%、100%，平均秩次分別為16.84、31.31、45.43，經(jīng)各組間兩兩比較進(jìn)行秩和檢驗(yàn)，其表達(dá)差異均有統(tǒng)計(jì)學(xué)意義（HC=24.63，P＜0.05）；IL-17R在減壓腦組織、低級(jí)別及高級(jí)別膠質(zhì)瘤中存在陽性表達(dá)，其陽性率分別為50%、83%、100%，平均秩次分別為18.42、35.73、49.65，經(jīng)各組間兩兩比較進(jìn)行秩和檢驗(yàn)，其表達(dá)差異均有統(tǒng)計(jì)學(xué)意義（HC=28.42，P＜0.05）。(2) RT-PCR方法檢測(cè)結(jié)果，IL-17mRNA在對(duì)照組減壓腦組織、低級(jí)別及高級(jí)別組膠質(zhì)瘤組織中表達(dá)的灰度值分別為31.62±1.95、85.79±4.58、134.77±6.32，， IL-17RmRNA表達(dá)為76.20±3.97、137.66±64、193.82±6.20，經(jīng)方差分析(F=221.04，p0.05)，且組間比較均有統(tǒng)計(jì)學(xué)差異（p0.05)。IL-17mRNA在對(duì)照組健康患者外周血、外傷減壓患者外周血、低級(jí)別及高級(jí)別組膠質(zhì)瘤患者外周血中表達(dá)的灰度值分別為42.09±1.65、48.21±1.79、103.71±4.37、184.67±6.51，IL-17RmRNA表達(dá)為83.18±4.57、89.47±3.82、158.45±5.34、237.32±7.29，經(jīng)方差分析(F=274.64，p0.05)，對(duì)照組與減壓組比較(t=-0.92，p0.05)，差異無統(tǒng)計(jì)學(xué)意義，余組間比較均有統(tǒng)計(jì)學(xué)差異（p0.05)。(3)Western blot方法檢測(cè)結(jié)果，IL-17mRNA在對(duì)照組減壓腦組織、低級(jí)別及高級(jí)別組膠質(zhì)瘤組織中表達(dá)的灰度值分別為0.23±0.01、0.42±0.01、0.67±0.02， IL-17RmRNA表達(dá)為0.34±0.01、0.68±0.02、0.89±0.03，經(jīng)方差分析(F=89.23，p0.05)，且組間比較均有統(tǒng)計(jì)學(xué)差異（p0.05)。IL-17蛋白質(zhì)在對(duì)照組健康患者外周血、外傷減壓患者外周血、低級(jí)別及高級(jí)別組膠質(zhì)瘤患者外周血中表達(dá)的灰度值分別為0.33±0.01、0.35±0.01、0.54±0.02、0.85±0.02，IL-17R蛋白質(zhì)的表達(dá)為0.42±0.01、0.43±0.01、0.78±0.02、0.94±0.03，經(jīng)方差分析(F=107.89，p0.05)，對(duì)照組與減壓組比較(t=-0.83，p0.05)，差異無統(tǒng)計(jì)學(xué)意義，余組間比較均有統(tǒng)計(jì)學(xué)差異（p0.05)。(4) IL-17在相同膠質(zhì)瘤患者外周血及相對(duì)應(yīng)的膠質(zhì)瘤組織中的表達(dá)明顯正相關(guān)(r=0.520，p=0.003)，IL-17R在相同膠質(zhì)瘤患者外周血及相對(duì)應(yīng)的膠質(zhì)瘤組織中的表達(dá)呈顯著正相關(guān)(r=0.636，p0.001)。結(jié)論：IL-17和IL-17R在不同級(jí)別膠質(zhì)瘤組織和外周血中的表達(dá)有差異，且與膠質(zhì)瘤的病理分級(jí)密切相關(guān)，隨著膠質(zhì)瘤級(jí)別的增高而增多、增強(qiáng)；相同膠質(zhì)瘤患者外周血及相對(duì)應(yīng)的膠質(zhì)瘤組織中的表達(dá)有明顯正相關(guān)；IL-17和IL-17R的表達(dá)可能是判斷膠質(zhì)瘤惡性程度的參考指標(biāo)。
[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.
【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R739.41

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本文編號(hào)：2250429