热休克蛋白70调节突触融合蛋白1在哮喘中的表达及意义

发布时间：2018-08-10 22:20

【摘要】：目的探讨热休克蛋白70（heat shock protein70, HSP70）、突触融合蛋白1（syntaxin1, SYX1）在哮喘病理生理过程中表达的变化及二者的相互作用；同时探讨地塞米松（Dexamethason，DXM）对哮喘模型大鼠肺组织中HSP70、SYX1表达的影响。方法30只健康Wistar大鼠按随机原则分为对照组，哮喘组，哮喘+地塞米松组，每组10只。通过卵清蛋白（ovalbumin，OVA）致敏、激发建立哮喘大鼠模型，采用免疫组织化学、逆转录PCR及蛋白免疫印迹检测三组间HSP70及SYX1基因和蛋白表达水平，免疫共沉淀技术检测HSP70与SYX1的相互关系。结果1.成功复制大鼠哮喘模型，哮喘组大鼠激发后开始烦躁不安，呼吸急促，腹式呼吸，哮鸣音，腹肌收缩和跌倒。对照组大鼠无明显上述表现，DXM组上述表现轻微。支气管肺组织切片HE染色显示哮喘组大鼠支气管、细支气管上皮下和小血管周围可见明显的炎症细胞浸润，气管上皮结构紊乱，部分脱落，管腔内渗出增加，，粘膜及粘膜下可见淋巴细胞，嗜酸性粒细胞为主的大量炎性细胞浸润；对照组大鼠气道粘膜未见明显水肿，粘膜结构完整，偶见少量炎症细胞浸润；地塞米松治疗组肺部炎症改变较哮喘组有所减轻。 2.通过免疫组化检测，光镜下见HSP70表达部位主要位于支气管上皮细胞，部分肺泡上皮细胞及炎症细胞， SYX1阳性细胞主要表达于支气管上皮细胞靠近支气管腔的包膜处。 3.与对照组比较，哮喘组HSP70及SYX1基因和蛋白表达水平均显著增加，差异有统计学意义（P＜0.05，P＜0.05）；与哮喘组比较，哮喘组+地塞米松组HSP70及SYX1基因与蛋白表达水平均显著降低，差异有统计学意义（P＜0.05，P＜0.05），但与正常对照组比较，差异均无统计学意义（P＞0.05，P＞0.05）。免疫共沉淀结果显示在哮喘大鼠肺组织中HSP70与SYX1存在相互作用。结论HSP70及SYX1在哮喘肺组织中表达水平均显著增高，HSP70通过调节SYX1表达水平及相互作用共同参与哮喘的发病机制，且其表达受地塞米松的抑制，为临床寻找药物干预新的靶点提供了理论依据。
[Abstract]:Objective to investigate the expression of heat shock protein 70 (HSP70) and syntaxin 1 (SYX1) in the pathophysiological process of asthma and their interaction, and to investigate the effect of dexamethasone on the expression of HSP70- SYX1 in lung tissue of asthmatic rats. Methods 30 healthy Wistar rats were randomly divided into control group and asthmatic dexamethasone group with 10 rats in each group. The asthmatic rat model was established by sensitizing ovalbumin OVA. The expression levels of HSP70 and SYX1 gene and protein were detected by immunohistochemistry, reverse transcription PCR and Western blot. The relationship between HSP70 and SYX1 was detected by immunoprecipitation. Result 1. The asthmatic rats in the asthma group were induced to become restless, shortness of breath, abdominal breathing, wheezing, abdominal muscle contraction and fall. The above findings in DXM group were slight. The HE staining of bronchopulmonary sections showed obvious inflammatory cell infiltration in the bronchioles, subcutaneous bronchioles and small blood vessels, disorder of tracheal epithelial structure, partial exudation, and increased exudation in the lumen of asthmatic rats. Lymphocytes were found in mucosa and submucous membrane and a large number of inflammatory cells were infiltrated mainly by eosinophils. In the control group there was no obvious edema in the airway mucosa and the mucosal structure was intact and a small number of inflammatory cells were occasionally infiltrated. The pulmonary inflammation in dexamethasone treatment group was less than that in asthma group. 2. 2. The expression of HSP70 was mainly located in bronchial epithelial cells, part of alveolar epithelial cells and inflammatory cells under light microscope, and SYX1 positive cells were mainly expressed in the capsule of bronchial epithelial cells near the bronchial cavity. 3. Compared with the control group, the expression levels of HSP70 and SYX1 gene and protein in asthma group were significantly increased (P < 0.05), and the expression levels of HSP70 and SYX1 gene and protein in asthmatic group were significantly lower than those in asthma group. The difference was statistically significant (P < 0.05 P < 0.05), but there was no significant difference compared with the control group (P > 0.05 P > 0.05). The results of immunoprecipitation showed that there was interaction between HSP70 and SYX1 in lung tissues of asthmatic rats. Conclusion the expression of HSP70 and SYX1 in asthmatic lung tissue is significantly increased. HSP70 is involved in the pathogenesis of asthma by regulating the expression and interaction of SYX1, and the expression of HSP70 is inhibited by dexamethasone. It provides a theoretical basis for finding new targets for drug intervention.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R562.25

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