慢性阻塞性肺疾病大鼠模型中骨骼肌蛋白降解机制的研究  

Skeletal muscle protein degradation mechanism in chronic obstructive pulmonary disease rats

作　　者：曾祥富[1] 刘朝晖[1] 梁志科[1] 范伟[1] 

机构地区：[1]广州医学院附属广州市第一人民医院呼吸内科,510180

出　　处：《国际呼吸杂志》2011年第23期1770-1776,共7页International Journal of Respiration

基　　金：基金项目：广东省自然科学基金项目（10151006001000012）;广州市卫生局资助项目（30504382207）

摘　　要：目的研究慢性阻塞性肺疾病（chronic obstructive pulmonary disease，COPD）大鼠模型的骨骼肌蛋白降解机制，为有效防治COPD患者骨骼肌蛋白高分解，功能障碍等难题提供科学理论和依据。方法成年雄性SD大鼠30只，分为模型组18只，对照组12只，采用反复熏香烟和气管内注入脂多糖复制COPD大鼠模型。酶联免疫吸附法（ELISA）测定血清和膈肌组织中的白介素6（IL-6），肿瘤坏死因子a（TNF—α ），γ干扰素（IFN- γ）等含量。蛋白免疫印迹法测定大鼠膈肌和腓肠肌内的E214K、MAFbx、MuRF-1的蛋白表达。结果在模型组中，血清和膈肌内的IL-6、TNF-a均较对照组显著升高，IFN- γ则较对照组显著降低。Western blot的结果显示：膈肌组织中E2—14K、MAFbx、MuRF-1均较对照组表达明显增强（各蛋白的相对表达量分别为0．96±0．12VS0．53±0．09、0．99±0．10VS0．53±0．08、0．95±0．08VS0．51±0．16，P〈0．01）。腓肠肌中E214K、MAFbx、MuRF-1亦较对照组表达增强（各蛋白的相对表达量分别为0．98±0．06VS0．83±0．05、1．00±0．04VS0．81±0．05、0．92±0．05VS0．71±0．05，P〈0．05）。体质量与TNF—α呈显著负相关（血清r=-0．84，膈肌r=-0．75，P值均〈0．01），与IFN-γ呈正相关（血清r=-0．71，膈肌r=-0．69，P值均〈0．01）；中性粒细胞百分比与IL-6呈正相关（血清r=0．82，膈肌r=0．91，P值均〈0．01）；膈肌组织中，IL-6、TNF—α的水平与膈肌的E2—14K、MAFbx、MuRF-1灰度值呈显著正相关，相关系数分别为r=0．86、r=0．94、r=0．89，P〈0．01和r=0．84、r=0．83、r=0．84，P〈0．01；IFN-γ与E2—14K、MAFbx、MuRF-1灰度值呈显著负相关，相关系数为r=0．81、r=0．80、r=0．78，P〈0．01。结论在COPD大鼠模型中，炎症因子、免疫调节因子的表达失衡和骨骼肌内泛素及泛素化蛋白的过度表达可能是引起骨骼肌蛋白降解加快的重�Objective To investgate protein degradation in skeletal muscle of chronic obstructive pulmonary disease （COPD） rats, in order to prevent increase of protein degradation, dysfunction and other problems in skeletal muscle of COPD patients,to provide scientific theories. Methods Thirty health male adult SD rats were randomly divided into two grops：a model group （ n = 18） and a normal control grop （ n = 12）. The COPD rat model was established by instillation of LPS and exposure to eigareete smoke for 28 days. The concentration of IL 6,TNF-α and IFN-γ in serum and diaphragm were measured by ELISA. The protein levels of E2- 14K, MAFbx and MuRF-1 in the diaphgrams and gastrocnemius were measured by Western blot. Results Compared with the normal control group, in the serum and the diaphragm, the IL-6, TNF-α were significantly higher than those of the control group, IFN-γ were significantly lower than the control group. Western blot result shows：in the diaphragm, compared with the normal control group,the protein expression of E2-14K, MAFbx and MuRF 1 increased significantly in the model group（0.96±0.12 vs 0. 53±0. 09,0. 99±0.10 vs 0.53±0.08 and 0.95±0.08 vs 0.51±0.16, P 〈0.01）,in the gastrocnemius, compared with the normal control group, the protein expression of E2 14K, MAFbx and MuRF-1 also increased significantly in the model group （0.98±0.06 vs 0. 83 ± 0. 05 , 1.00±0.04 vs 0.81±0.05 and 0.92±0.05 vs 0. 71±0. 05, P 〈0.05）. Weight of rats with TNF-a was strong negatively correlated（serum r = 0.8, diaphragm r = -0.75, P 〈0.01） and with IFN-γ was significant positively correlated （serum r = 0.82, diaphragm r = 0.91, P 〈0.01 ）. In diaphragm, IL-6, TNF-a level and E2-14K, MAFbx,MuRF-1 gray value was significantly positively correlated （ r = 0.86, r = 0.94, r =0.89, P 〈0.01 and r =0.84, r =0.83, r =0.84, P 〈0.01）;IFN-γ and E2-14K,MAFbx, MuRF-1 gray value was significantly negatively correlated （ r = 0.81, r = 0.80, r = 0.78, P 〈 0.01 ）. Conclusions Infl

关 键 词：慢性阻塞性肺疾病 骨骼肌 蛋白降解 白介素6 肿瘤坏死因子Α Γ干扰素 

分 类 号：R563.9[医药卫生—呼吸系统]