呼吸道合胞病毒感染对肺组织GRK2的影响及其与哮喘的关系
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徐州医学院附属医院课题资助


The influence of respiratory syncytial virus infection on GRK2 expression of lung tissue and its relationship with asthma
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    目的:探讨呼吸道合胞病毒(respiratory syncytial virus,RSV)感染鼠肺组织后,G蛋白偶联受体激酶2(G protein coupled receptor kinase 2,GRK2)的变化规律及其与哮喘的关系-方法:Balb/c雌性小鼠100只,随机分为5组,每组20只-正常对照组,于第1-14天予生理盐水0.2 ml腹腔注射,第21~25天用37℃生理盐水20 ml雾化吸入30 min;RSV感染组,于第19-20-21天,以RSV按浓度1 × 106 PFU-100 -滋l/次鼻腔滴入;哮喘组,第1-14天予以鸡卵白蛋白(ovalbumin,OVA)100 μg-AL(OH)3 2 mg腹腔注射,第21天开始激发,以2%OVA生理盐水20 ml雾化吸入30 min,持续激发5 d;双重感染组,模型制备同哮喘组,第19-20-21天,以RSV按浓度1 × 106 PFU-100 -滋l/次鼻腔滴入;地塞米松干预组,RSV感染同RSV组,第21~25天予腹腔注入地塞米松0.2 mg/(kg-d)-末次激发后24 h,10%水合氯醛腹腔注射麻醉(0.3 ml/kg),处死小鼠,取左肺上叶和右肺中叶,4%甲醛固定,分别用于免疫荧光检测GRK2的表达和组织病理改变检测;取右肺下叶用于Western blot检测各组GRK2的表达;无菌条件下取左肺下叶,用于肺组织病毒分离及免疫荧光鉴定-结果:RSV组GRK2表达较正常组显著增加,与哮喘组相比差异有统计学意义;双重感染组GRK2表达较RSV组显著增加;地塞米松干预组GRK2表达较RSV组显著减少-结论:RSV感染小鼠肺组织后GRK2表达增加,合并哮喘时表达显著增加,地塞米松干预对GRK2的表达有一定的抑制作用-

    Abstract:

    Objective:To investigate the G protein coupled receptor kinase 2 (GRK2) changes and its relationship with asthma after respiratory syncytial virus (RSV) infection in lung tissue of mice. Methods:A total of 100 female Balb/c mice were randomly divided into 5 groups with 20 each group. In the normal control group,intraperitoneal injection of 0.2 ml saline were performed at day 1 and 14,and inhalation of 20 ml atomized saline at 37℃ were taken for 30 min at day 21~25. In the RSV group,RSV of 1×106 pfu in consistency and 100 -滋l/time in volume were dropped into the nasal cavity at day 19,20 and 21. In the asthma group,intraperitoneal injection of OVA/AL (OH)3 (OVA 100 μg,AL (OH)3 2 mg) were performed at day 1 and 14,and inhalation of 2%OVA saline of 20 ml were taken for 30 min at day 21 and the excitation was lasted for 5 days. In the double infection group,model preparation was the same with the asthma group,but RSV of 1 × 106 pfu in consistency and 100 -滋l/time in volume were dropped into the nasal cavity at day 19,20 and 21. In the dexamethasone intervention group,RSV infection was the same to the RSV group,but intraperitoneal injection of dexamethasone 0.2 mg/(kg-d) were performed at day 21 to 25. Twenty-four hours after the last excitation,by anesthesia with 10% chloral hydrate intraperitoneal injection (0.3 ml/kg),the mice were sacrificed. The left upper lobe and right middle lobe of the lung,4% formaldehyde fixed,were used for expression and pathological immunofluorescence detection of GRK2 change detection. The lower lobe of the right lung was performed to detected GRK2 expression of each group by Western blot. Under the sterile conditions,the lower lobe of the left lung was taken for virus isolation and immunofluorescence identification. Results:In the RSV group,GRK2 expression was significantly increased compared with the normal group,and showed significant differences compared with the asthma group. In the double infection group,GRK2 expression was significantly increased compared with the RSV group. In the dexamethasone intervention group,GRK2 expression was significantly reduced compared with the RSV group. Conclusion:The expression of GRK2 was increased after the RSV infection in lung tissue of the mice,and significantly increased in terms to asthma. The dexamethasone intervention inhitits the expression of GRK2.

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吴 铭,张 华,武 怡,屈昌雪,李 刚.呼吸道合胞病毒感染对肺组织GRK2的影响及其与哮喘的关系[J].南京医科大学学报(自然科学版),2014,(11):1463-1468

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  • 收稿日期:2014-03-13
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  • 在线发布日期: 2014-11-21
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