论文部分内容阅读
目的研究慢性阻塞性肺疾病(COPD)大鼠膈肌萎缩信号通路——泛素-蛋白酶体通路中E2-14K、MAFbx和MuRF-1蛋白表达及核转录因子κB(NF-κB)p50表达情况,以及前炎症因子白细胞介素17(IL-17)在大鼠血清及膈肌中的表达,探讨膈肌萎缩的可能调控机制。方法采用气管内滴入脂多糖+反复熏香烟法复制COPD大鼠动物模型。Western blot法测定大鼠膈肌内E2-14K、MAFbx、MuRF-1及NF-κB p50表达,ELISA法测定大鼠血清及膈肌中IL-17的表达。结果 Westernblot结果显示COPD大鼠膈肌中E2-14K、MAFbx、MuRF-1及NF-κB p50蛋白的表达均较对照组明显升高(0.96±0.12比0.53±0.09,0.99±0.10比0.53±0.08,0.95±0.08比0.51±0.16,1.11±0.10比0.64±0.50,P均<0.01)。ELISA结果显示IL-17在血清及膈肌中的表达均升高(P均<0.01)。NF-κB p50与E2-14K、MAFbx、MuRF-1水平呈显著正相关(r值分别为0.82、0.92、0.86,P均<0.01);血清IL-17与中性粒细胞百分比、NF-κB p50、E2-14K、MAFbx、MuRF-1呈显著正相关(r值分别为0.94、0.90、0.85、0.84及0.79,P均<0.01);膈肌IL-17与中性粒细胞百分比、NF-κB p50、E2-14K、MAFbx、MuRF-1呈显著正相关(r值分别为0.88、0.90、0.72、0.86及0.80,P均<0.01);血清IL-17与膈肌IL-17呈显著正相关(r=0.84,P<0.01)。结论 COPD大鼠膈肌中泛素-蛋白酶体通路与NF-κB通路表达上调可能是导致膈肌萎缩的主要原因之一,IL-17可能参与了膈肌萎缩的调控。
Objective To investigate the expression of E2-14K, MAFbx and MuRF-1 and the expression of NF-κB p50 in the diaphragm atrophy signal pathway-ubiquitin-proteasome pathway in chronic obstructive pulmonary disease (COPD) As well as the expression of interleukin 17 (IL-17) in rat serum and diaphragmatic muscle, and explore the possible regulatory mechanism of diaphragmatic atrophy. Methods COPD rat model was established by intratracheal instillation of lipopolysaccharide + repeated cigarette smoking. The expressions of E2-14K, MAFbx, MuRF-1 and NF-κB p50 in the diaphragm were measured by Western blot. The expression of IL-17 in serum and diaphragm of rats was measured by ELISA. Results The results of Western blot showed that the expressions of E2-14K, MAFbx, MuRF-1 and NF-κB p50 in diaphragmatic muscle of COPD rats were significantly higher than those in control group (0.96 ± 0.12 vs 0.53 ± 0.09, 0.99 ± 0.10 vs 0.53 ± 0.08, 0.95 ± 0.08 vs. 0.51 ± 0.16, 1.11 ± 0.10 vs. 0.64 ± 0.50, P <0.01). ELISA results showed that the expression of IL-17 in serum and diaphragm increased (P <0.01). There was a significant positive correlation between NF-κB p50 and E2-14K, MAFbx and MuRF-1 (r = 0.82,0.92,0.86, P <0.01 respectively). The percentage of serum IL-17 to neutrophil, NF- p50, E2-14K, MAFbx and MuRF-1 (r values were 0.94,0.90,0.85,0.84 and 0.79 respectively, all P <0.01). The percentage of IL-17 and neutrophil in diaphragm, NF-κB p50, E2-14K, MAFbx and MuRF-1 (r values were 0.88,0.90,0.72,0.86 and 0.80 respectively, P <0.01), and there was a significant positive correlation between serum IL-17 and diaphragmatic muscle IL-17 r = 0.84, P <0.01). Conclusion Upregulation of ubiquitin-proteasome pathway and NF-κB pathway in the diaphragmatic muscle of COPD rats may be one of the main causes of diaphragmatic atrophy. IL-17 may be involved in the regulation of diaphragmatic atrophy.