IL-6基因的表达及其多态性与变应性鼻炎的相关性研究

发布时间：2018-07-05 13:49

本文选题：变应性鼻炎 + IL-6　；参考：《山东大学》2016年博士论文

【摘要】：目的目前已有多个研究发现,白介素6(IL-6)基因多态性是多种免疫相关疾病的易感基因。最近的研究显示,在过敏性呼吸系统疾病中,IL-6担当重要角色。有关数据显示,IL-6的表达同变应性鼻炎的发病之间存在一定的关系,本研究通过三部分实验来探讨IL-6基因多态性与中国汉族人群变应性鼻炎易感性之间是否相关性。方法(1)采用HE染色和免疫组织化学法分别检测45例变应性鼻炎患者下鼻甲组织、15例鼻中隔偏曲患者下鼻甲组织(对照组)的病理组织学变化以及IL-6的表达。(2)收集265例变应性鼻炎患者的外周静脉血及265例对照组的外周静脉血样本,对两组样本进行病例对照研究。提取血液样本中的DNA,并对DNA样本中IL-6基因的两个多态位点(rs1800795和rs1800796)进行基因分型,探讨IL-6的基因多态性与变应性鼻炎易感性的关系。基因分型采取PCR-RFLP和直接测序的法。(3) 利用Balb/c小鼠建立卵清蛋白致敏的变应性小鼠模型,按末次激发后处死时间分为6组(30min、1h、2h、6h、12h、24h),同时使用生理盐水作为对照组,每组各5只小鼠。取各组小鼠鼻黏膜组织,利用HE染色法检测各组小鼠鼻黏膜组织的病理组织学变化情况,western blot法检测IL-6在各组小鼠鼻黏膜组织中的表达情况。结果(1)HE染色结果表明,变应性鼻炎患者的鼻黏膜上皮组织发生部分脱落,视野下能够见到杯状细胞增生,组织存在水肿现象,腺体增生,血管扩张,在上皮组织间可见到医以嗜酸性粒细胞和淋巴细胞为主的大量炎细胞浸润,其中还包括嗜碱性粒细胞、中性粒细胞等。对两组组织切片中嗜酸性细胞进行计数,结果表明变应性鼻炎组嗜酸性粒细胞浸润显著,为(31.92±2.95),正常对照组为(3.83±2.03),变应性鼻炎组明显高于正常对照组,两者相比差异显著(P=0.0010.05); western blot检测结果表明,IL-6蛋白在变应性鼻炎患者鼻黏膜组织中的相对表达量为(0.40±0.16),而在正常对照组下鼻甲组织中的相对表达量为(0.26±0.14),变应性鼻炎组明显高于正常对照组(P=0.0030.05)。(2)对IL-6基因多态性进行分析,结果表明IL-6 rs1800795等位基因频率在变应性鼻炎患者与对照组中存在显著差异(χ2=4.52,P=0.03)；多因素logistic回归分析表明,相较于G等位基因,IL-6rs1800795的C等位基因具有更高的变应性鼻炎发病风险,OR值为1.31(1.01～1.68)。(3) 动物行为学观察结果表明,变应性鼻炎小鼠模型造模成功；HE染色结果表明,各变应性鼻炎模型组小鼠的鼻腔黏膜上皮均出现不同程度的结构紊乱,同时还可见充血、水肿以及黏膜层增厚等现象,此外还伴有黏膜上皮组织坏死以及脱落等,纤毛上皮可见有融合现象出现,细胞核发生形变,形状不规则,柱状细胞以及杯状细胞增生,血管扩张并增生,大量肥大细胞、中性粒细胞以及嗜酸性粒细胞、淋巴细胞浸润。Western blot检测结果表明,各组小鼠鼻黏膜组织中均有IL-6蛋白的表达,其中在30 min组中的相对表达量为0.142±0.064,在1h组中的相对表达量为0.214±0.104,在2h组中相对表达量为0.274±0.092,在6h组中的相对表达量为0.324±0.122,然后随着时间的延长呈逐渐下降趋势,在12h组中的相对表达量为0.236+0.059,在24h组中的表达量为0.172±0.058。与对照组的相对表达量0.068±0.020相比,实验组小鼠鼻黏膜组织中IL-6蛋白的表达量较正常组增高。结论IL-6在变应性鼻炎患者鼻黏膜组织中呈高表达,且变应性鼻炎的易感性与IL-6基因多态性显著相关。
[Abstract]:Objective a number of studies have found that the interleukin 6 (IL-6) gene polymorphism is a susceptible gene for a variety of immune related diseases. Recent studies have shown that IL-6 plays an important role in allergic respiratory diseases. The relevant data show that there is a relationship between the expression of IL-6 and the incidence of allergic rhinitis. This study is conducted through three parts of the study. The correlation between IL-6 gene polymorphism and allergic rhinitis susceptibility in Chinese Han population was investigated. Methods (1) the histopathological changes in inferior turbinate tissue of 45 patients with allergic rhinitis and 15 cases of nasal septum deviation patients were detected by HE staining and immunohistochemistry, and the table of IL-6 (2) collect the peripheral venous blood of 265 patients with allergic rhinitis and 265 cases of the peripheral venous blood samples from the control group, carry out a case-control study on the two groups of samples, extract the DNA in the blood samples, and classify the two polymorphic loci of the IL-6 gene (rs1800795 and rs1800796) in the DNA sample, and explore the gene polymorphism and allergy of the IL-6. The relationship between the susceptibility of sexual rhinitis. The genotyping was made by PCR-RFLP and direct sequencing. (3) the allergic mice induced by ovalbumin were established by Balb/c mice. The time of death was divided into 6 groups (30min, 1H, 2h, 6h, 12h, 24h), and the physiological salt water was used as the control group, and 5 mice in each group were taken as the control group, and the nasal mucosa group of each group was taken. The pathological changes of nasal mucosa in each group were detected by HE staining. The expression of IL-6 in nasal mucosa of each group was detected by Western blot. Results (1) the results of HE staining showed that the nasal epithelium of the allergic rhinitis patients were separated and the goblet cell hyperplasia could be seen in the field of vision. There is edema, hyperplasia of glands and dilatation of blood vessels. A large number of inflammatory cells, mainly eosinophils and lymphocytes, are seen between the epithelial tissues, including basophils and neutrophils. The eosinophils in the two groups of tissue sections are counted, and the results show eosinophilic granulocyte immersion in allergic rhinitis group. It was significant (31.92 + 2.95), the normal control group was (3.83 + 2.03), the allergic rhinitis group was significantly higher than the normal control group, and the difference was significant (P=0.0010.05). The results of Western blot detection showed that the relative expression of IL-6 protein in the nasal mucosa of patients with allergic rhinitis was (0.40 + 0.16), but in the normal control group, the inferior turbinate tissue was in the normal control group. The relative expression amount was (0.26 + 0.14), the allergic rhinitis group was significantly higher than that of the normal control group (P=0.0030.05). (2) the IL-6 gene polymorphism was analyzed. The results showed that the IL-6 rs1800795 allele frequency was significantly different in the allergic rhinitis patients and the control group (x 2= 4.52, P=0.03), and the multiple factor Logistic regression analysis showed that it was compared with G. The allele of the allele, IL-6rs1800795 C allele had a higher risk of allergic rhinitis, the OR value was 1.31 (1.01 ~ 1.68). (3) the animal behavior observation showed that the model of allergic rhinitis mice model was successful, and the HE staining results showed that the nasal mucosa epithelium of each allergic rhinitis model group had different degree of structural turbulence. At the same time, hyperemia, edema, and thickening of mucous membrane were also seen, in addition to necrosis and exfoliation of epithelial tissue of mucous membrane. Ciliated epithelium showed fusion phenomenon, nuclear deformation, irregular shape, columnar cells and goblet cells hyperplasia, vasodilatation and hyperplasia, large numbers of mast cells, neutrophils, and neutrophils. The results of.Western blot detection in acid granulocyte and lymphocyte infiltration showed that the expression of IL-6 protein in the nasal mucosa of each group was 0.142 + 0.064 in the 30 min group, the relative expression in the 1H group was 0.214 + 0.104, the relative expression in the 2H group was 0.274 + 0.092, and the relative expression in the 6h group was 0.3. 24 + 0.122, and then gradually decreasing with time, the relative expression in 12h group was 0.236+0.059, the expression amount in 24h group was 0.172 + 0.058. and the relative expression of control group was 0.068 + 0.020. The expression of IL-6 protein in the nasal mucosa of the experimental group was higher than that of the normal group. Conclusion IL-6 is in allergic rhinitis. The expression of IL-6 gene was highly expressed in nasal mucosa, and the susceptibility of allergic rhinitis was significantly correlated with the polymorphism of the gene.
【学位授予单位】：山东大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R765.21

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