BATF、RORγt及IL-17在哮喘小鼠脾组织中的表达及其意义

发布时间：2018-06-13 18:14

本文选题：B细胞活化转录因子 + ROR　；参考：《泸州医学院》2013年硕士论文

【摘要】：目的：观察B细胞活化转录因子(B-cell activiting transcription factor, BATF)、维甲酸相关孤核受体(retinoic acid-related orphan recepptor γt, ROR γ t白细胞介素-17(interleukin-17, IL-17)在哮喘小鼠脾脏组织中的表达,探讨BATF在哮喘发病机制中的作用,为哮喘治疗寻找新途径。方法：取健康雌性BALB/c小鼠15只,随机分为正常对照(NS)组、哮喘(AS)组、地塞米松(DXM)组,每组小鼠均为5只。AS组、DXM组小鼠分别于第0、7、14天用卵清蛋白(OVA)100ug与氢氧化铝(Al (OH) 3) 1mg混合的生理盐水混悬液0.2ml腹腔内注射致敏,NS组则用等量生理盐水代替行腹腔注射；第21天开始NS组予以生理盐水,AS组及DXM组予1%的OVA生理盐水溶液行超声雾化吸入,每天1次,每次40分钟,连续5天进行激发；DXM组于每次激发前30分钟,予地塞米松磷酸钠注射液1.0mg/kg腹腔注射,NS组及AS组则予以等量的生理盐水腹腔注射,时间同上。所有小鼠都于末次激发24小时后处死。收集支气管肺泡灌洗液(BALF)行白细胞计数及分类计数；肺组织病理切片HE染色来评估小鼠气道及肺组织炎性细胞浸润情况；酶联免疫吸附法(ELISA)检测脾脏组织匀浆IL-17的浓度；real-time PCR法检测脾脏组织中BATF mRNA、ROR γ t mRNA的表达水平。结果：(1)肺组织病理学HE染色：AS组可见小气道及小血管周围大量的炎性细胞浸润,粘膜及粘膜下层水肿,肺泡腔内可见嗜酸性粒细胞、中性粒细胞、单核细胞及溢出的红细胞；DXM组炎细胞、渗出物较AS组少,支气管上皮基本完整,可见明显的炎症吸收；NS组小气道粘膜无明显水肿,管腔光滑、无闭塞,周围小血管基本未见炎性细胞的浸润。(2)与NS组比较,AS组、DXM组小鼠BALF中白细胞总数、中性粒细胞数和嗜酸性粒细胞数明显增多(P0.05),与AS组比较,DXM组BALF中白细胞总数、中性粒细胞数和嗜酸性粒细胞数有明显降低(P0.05)。(3)脾脏组织匀浆IL-17的浓度：AS组IL-17浓度最高,DXM组其次,NS组最低,三组之间的差异有统计学意义(P0.05),且与NS组比较,AS组及DXM组IL-17浓度均明显增高,差异有统计学意义(P0.05),DXM组较AS组IL-17浓度降低,差异有统计学意义(P0.05)。(4)脾脏组织中BATF mRNA.ROR γ t mRNA表达水平：AS组水平最高,DXM组其次,NS组最低,三组之间的差异有统计学意义(P0.05),且与NS组比较,AS组及DXM组BATF mRNA. ROR γ t mR NA表达水平均明显增高,差异有统计学意义(P0.05),DXM组较AS组BATF mRNA、ROR y t mRNA表达水平降低,差异有统计学意义(P0.05)。(5)相关分析表明：脾脏组织中IL-17的浓度与BALF中白细胞总数、中性粒细胞数、嗜酸性粒细胞数成正相关(r=0.838,0.737,0.882,P均0.005)；脾脏组织BATF mRNA的表达量与ROR γ t mRNA表达量、脾组织匀浆IL-17浓度呈正相关(r=0.807,0.833,P均0.005)：脾组织匀浆IL-17浓度与脾脏组织ROR γ t mRNA表达量呈正相关,(r=0.890,P0.005)。结论：(1)BATF与ROR γ t共同调节IL-17的分泌,并通过IL-17的致炎作用参与哮喘气道炎症的发生发展,(2)地塞米松可能通过BATF、ROR γ t途径部分阻止脾源性Th17细胞的分化,减少IL-17的分泌,从而改善哮喘气道炎症细胞的浸润,缓解哮喘症状。
[Abstract]:Objective: To observe the expression of B cell activation transcription factor (B-cell activiting transcription factor, BATF), retinoic acid related soliton receptor (retinoic acid-related orphan recepptor gamma T, ROR gamma) in the spleen tissues of asthmatic mice and to explore the role of asthma in the pathogenesis of asthma. Methods: 15 healthy female BALB/c mice were randomly divided into normal control (NS) group, asthma (AS) group and dexamethasone (DXM) group, each group was 5.AS groups, and DXM mice were injected intraperitoneally with the mixture of ovalbumin 100ug (OVA) 100ug and aluminum hydroxide (Al (OH) 3) on day 0,7,14. Group NS was injected with equal physiological saline instead of intraperitoneal injection; group NS was given physiological saline at twenty-first days, group AS and group DXM were given 1% OVA physiological saline solution by ultrasonic atomization inhalation, 1 times a day, 40 minutes each time for 5 days, and 30 minutes before each stimulation, group DXM was given to Dexamethasone Sodium Phosphate Injection 1.0mg/kg abdominal cavity. Injection, group NS and group AS were injected with equal amount of normal saline. All mice were killed after 24 hours of last excitation. Bronchoalveolar lavage fluid (BALF) was collected to count the white blood cell count and classified count; lung tissue pathological section HE staining was used to evaluate the infiltration of inflammatory cells in the airway and lung tissues; enzyme linked immunosorbent assay. The concentration of IL-17 in the spleen tissue homogenate was detected by adsorption method (ELISA); the expression level of BATF mRNA and ROR t mRNA in the spleen tissues was detected by real-time PCR. Results: (1) HE staining of lung histopathology: AS group could see large number of inflammatory cell infiltration in small airway and small vessels, mucous membrane and submucosa edema, and eosinophil in the alveoli. Cells, neutrophils, mononuclear cells and spillover red cells, group DXM inflammatory cells, exudates less than group AS, bronchial epithelium was basically complete, visible inflammatory absorption, NS group small airway mucosa without obvious edema, smooth lumen, no occlusion, surrounding small vascular basically no infiltration of inflammatory cells. (2) compared with group NS, AS group, group DXM mice B The total number of leucocytes and neutrophils and eosinophils in ALF increased significantly (P0.05). Compared with the AS group, the total number of white blood cells, neutrophils and eosinophils in group DXM were significantly decreased (P0.05). (3) the concentration of IL-17 in the spleen tissue homogenate was the highest in the AS group, the second in the DXM group, the lowest in the NS group and the difference between the three groups. The difference was statistically significant (P0.05), and compared with the NS group, the IL-17 concentration in AS and DXM groups increased significantly (P0.05), and the concentration of IL-17 in group DXM was lower than that in AS group, and the difference was statistically significant (P0.05). (4) the level of BATF mRNA.ROR gamma expression in the spleen tissue was the highest, followed by the group, the lowest, three groups. The difference was statistically significant (P0.05), and compared with the NS group, the BATF mRNA. ROR gamma t mR NA expression level in the group AS and the DXM group was significantly higher, and the difference was statistically significant (P0.05). (5) the correlation analysis showed that the concentration of the spleen in the spleen tissue and the concentration of the spleen T The total number of leucocytes, neutrophils and eosinophils were positively correlated (r=0.838,0.737,0.882, P 0.005); the expression of BATF mRNA in the spleen tissue was positively correlated with the ROR gamma t mRNA expression, and the concentration of IL-17 in the spleen tissue homogenate was positively correlated (r=0.807,0.833, P 0.005): the IL-17 concentration of spleen tissue homogenization and the expression of splenic tissue ROR gamma expression Cheng Zhengxiang Guan, (r=0.890, P0.005). Conclusion: (1) BATF and ROR gamma t regulate the secretion of IL-17, and participate in the development of airway inflammation through the inflammatory action of IL-17. (2) dexamethasone may pass BATF, ROR gamma t pathway partially prevent the differentiation of spleen derived Th17 cells, reduce the secretion of IL-17, and thus improve the infiltration of airway inflammatory cells in asthma. Relieve the symptoms of asthma.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R562.25

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