發(fā)布時間：2018-08-10 22:20

【摘要】：目的探討熱休克蛋白70（heat shock protein70, HSP70）、突觸融合蛋白1（syntaxin1, SYX1）在哮喘病理生理過程中表達的變化及二者的相互作用；同時探討地塞米松（Dexamethason，DXM）對哮喘模型大鼠肺組織中HSP70、SYX1表達的影響。 方法30只健康Wistar大鼠按隨機原則分為對照組，哮喘組，哮喘+地塞米松組，每組10只。通過卵清蛋白（ovalbumin，OVA）致敏、激發(fā)建立哮喘大鼠模型，采用免疫組織化學、逆轉錄PCR及蛋白免疫印跡檢測三組間HSP70及SYX1基因和蛋白表達水平，免疫共沉淀技術檢測HSP70與SYX1的相互關系。 結果1.成功復制大鼠哮喘模型，哮喘組大鼠激發(fā)后開始煩躁不安，呼吸急促，腹式呼吸，哮鳴音，腹肌收縮和跌倒。對照組大鼠無明顯上述表現(xiàn)，DXM組上述表現(xiàn)輕微。支氣管肺組織切片HE染色顯示哮喘組大鼠支氣管、細支氣管上皮下和小血管周圍可見明顯的炎癥細胞浸潤，氣管上皮結構紊亂，部分脫落，管腔內滲出增加，，粘膜及粘膜下可見淋巴細胞，嗜酸性粒細胞為主的大量炎性細胞浸潤；對照組大鼠氣道粘膜未見明顯水腫，粘膜結構完整，偶見少量炎癥細胞浸潤；地塞米松治療組肺部炎癥改變較哮喘組有所減輕。 2.通過免疫組化檢測，光鏡下見HSP70表達部位主要位于支氣管上皮細胞，部分肺泡上皮細胞及炎癥細胞， SYX1陽性細胞主要表達于支氣管上皮細胞靠近支氣管腔的包膜處。 3.與對照組比較，哮喘組HSP70及SYX1基因和蛋白表達水平均顯著增加，差異有統(tǒng)計學意義（P＜0.05，P＜0.05）；與哮喘組比較，哮喘組+地塞米松組HSP70及SYX1基因與蛋白表達水平均顯著降低，差異有統(tǒng)計學意義（P＜0.05，P＜0.05），但與正常對照組比較，差異均無統(tǒng)計學意義（P＞0.05，P＞0.05）。免疫共沉淀結果顯示在哮喘大鼠肺組織中HSP70與SYX1存在相互作用。 結論HSP70及SYX1在哮喘肺組織中表達水平均顯著增高，HSP70通過調節(jié)SYX1表達水平及相互作用共同參與哮喘的發(fā)病機制，且其表達受地塞米松的抑制，為臨床尋找藥物干預新的靶點提供了理論依據(jù)。
[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.
【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R562.25

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