β-烯醇化酶及其mRNA在COPD下肢骨骼肌表达水平的研究

Beta-enolase and m RNA expression levels in quadriceps femoris muscle of COPD patients

目的探讨两种不同临床表型(骨骼肌萎缩组和非萎缩组)COPD,其下肢骨骼肌β-烯醇化酶蛋白及m RNA表达情况。方法收集昆明医科大学第一附属医院骨科手术诊疗的37例病人股四头肌标本,正常对照组11例、COPD患者26例,根据患者体质指数(BMI)、去脂肪指数结果,将COPD患者分为骨骼肌萎缩组(萎缩组)和骨骼肌非萎缩组(非萎缩组),萎缩组12例:男7例,女5例,平均(71.6±11.6)岁;非萎缩组14例:男6例,女8例,平均(72.4±8.8)岁。对照组男6例,女5例,平均(71.0±4.0)岁。利用蛋白质印迹法定量测定病人股四头肌β-烯醇化酶蛋白浓度,然后利用荧光定量PCR法检测病人股四头肌β-烯醇化酶m RNA转录水平,分析比较三组病人股四头肌中β-烯醇化酶m RNA转录水平及蛋白表达差异。结果萎缩组、非萎缩组、正常对照组中β-烯醇化酶蛋白及m RNA表达均具有统计学意义(P<0.05),非萎缩组β-烯醇化酶蛋白表达高于正常对照组(1.000±0.072)(P值0.0005),萎缩组β-烯醇化酶蛋白表达(1.200±0.078)高于非萎缩组(1.030±0.075)(P值0.0001)。非萎缩组β-烯醇化酶m RNA表达高于正常对照组(1.000±0.115)(P值0.0087),萎缩组β-烯醇化酶m RNA表达(2.600±0.947)高于非萎缩组(1.39±0.529)(P值0.0001)。萎缩组及非萎缩组中β-烯醇化酶蛋白浓度及β-烯醇化酶m RNA表达水平一致升高,但两组中β-烯醇化酶蛋白浓度与β-烯醇化酶m RNA表达比例关系不一致。结论 COPD骨骼肌萎缩组和COPD骨骼肌非萎缩组,其下肢骨骼肌β-烯醇化酶的表达存在差异。

Objective To explore the expression status of beta-enolase（β-enolase） from quadriceps femoris muscle in two kinds of muscle clinical phenotype（skeletal muscle atrophy group and skeletal muscles non-atrophy group） of chronic obstructive pulmonary disease（COPD）. Methods 37 inpatients from Orthopedics Department of the First Affiliated Hospital of Kunming Medical University, were divided into 11 patients without COPD and 26 patients with COPD, in addition, according to body mass index, fat free mass index and quadriceps cross-section diameter,patients with COPD were divided into 14 skeletal muscles non-atrophy patients（SMNA） and 12 skeletal muscle atrophy patients（SMA）.β-enolase concentration of femoris quadriceps specimens was quantitatively determined using Western Blot methods, β-enolase m RNA expression levels of femoris quadriceps specimens were also quantitatively measured using RT-PCR methods, then compared expression levels differences between each group.Results There was significant difference in β-enolase quantitative concentration and β-enolase m RNA expression level in each groups（P〈0.05）, further more, β-enolase concentration expression level was significantly higher（P=0.0001） in SMA group（1.200±0.078） than in SMNA group（1.03±0.075）, β-enolase concentration expression level was significantly higher（P=0.0005） that in SMNA group than in the control group（1.000 ±0.072）. β-enolasem RNA expression level was significantly higher（P=0.0087） in SMNA group than that in the control group（1.000±0.115）. β-enolasem RNA expression level was significantly higher（P=0.0001） in SMA group（2.600 ±0.947） than that in SMNA group（1.390 ±0.529）. β-enolase concentration and β-enolase m RNA expression level increased at equal pace in SMNA group and SMA group,however, β-enolase quantitative concentration and β-enolasem RNA expression level were inconsistent in the two groups. Conclusion The expression status of β-enolase in quadriceps femoris muscle was different in two kinds of muscle clinical phenotype of COPD.