β-烯醇化酶及mRNA在COPD患者下肢骨骼肌表达水平研究
发布时间：2019-05-20 02:29
【摘要】：目的：测定p-烯醇化酶蛋白及其mRNA水平在正常对照组、COPD非萎缩组、COPD萎缩组表达情况,探讨p-烯醇化酶在两种不同表型(萎缩型和非萎缩型)COPD的调控机制。 方法：收集昆明医科大学第一附属医院骨科手术诊疗患者股四头肌标本,其中正常对照组11例,COPD非萎缩组14例,COPD萎缩组12例。采用RT-PCR测定37例患者β-烯醇化酶mRNA水平,采用Western Blot技术检测37例患者β-烯醇化酶蛋白,分析比较3组患者股四头肌p-烯醇化酶mRNA转录水平及蛋白表达差异,结合患者去脂肪指数及股四头肌周径等影响因素,探讨β-烯醇化酶在两种不同表型(萎缩型和非萎缩型)COPD的调控机制。 结果：1.β-烯醇化酶蛋白相对表达量：①与正常组比较,COPD骨骼肌萎缩组患者股四头肌中β-烯醇化酶蛋白表达显著升高(P0.05)：②与正常组比较,COPD骨骼肌非萎缩组患者股四头肌p-烯醇化酶蛋白表达虽然有升高趋势,但差异不显著(P0.05)；③与COPD骨骼肌萎缩组比较,COPD骨骼肌非萎缩组患者股四头肌β-烯醇化酶蛋白表达无差异(P0.05)。 2.p-烯醇化酶mRNA的相对表达量(2-ΔΔCt):COPD骨骼肌萎缩组、COPD骨骼肌非萎缩组、正常对照组患者股四头肌β-烯醇化酶mRNA表达具有差异性(P0.05),进一步两两比较,各组间差异均有统计学意义,COPD骨骼肌萎缩组中p-烯醇化酶表达明显高于COPD骨骼肌非萎缩组、正常对照组,COPD骨骼肌非萎缩组p-烯醇化酶表达高于正常对照组。 3.COPD骨骼肌萎缩组p-烯醇化酶蛋白浓度与p-烯醇化酶mRNA表达水平一致升高,COPD骨骼肌非萎缩组骨骼肌中p-烯醇化酶蛋白浓度与p-烯醇化酶mRNA表达水平呈升高趋势。 结论：1.COPD骨骼肌萎缩组患者股四头肌β-烯醇化酶蛋白浓度及mRNA水平较正常组高,提示COPD骨骼肌萎缩患者p-烯醇化酶表达上调,可能与COPD存在慢性缺氧、骨骼肌类型转换有关； 2.COPD骨骼肌萎缩组患者p-烯醇化酶蛋白浓度及p-烯醇化酶mRNA水平较COPD骨骼肌非萎缩组高,p-烯醇化酶蛋白浓度及p-烯醇化酶mRNA水平在两种骨骼肌表型之间存在差异,表明可能不同的调控机制在两种表型对p-烯醇化酶的上调发挥了不同作用； 3.萎缩组及非萎缩组骨骼肌p-烯醇化酶蛋白浓度与p-烯醇化酶nRNA水平有一定的正相关性：①萎缩组患者通过增加β-烯醇化酶mRNA的量使p-烯醇化酶蛋白增高,说明DNA层面的调控在p-烯醇化酶的上调中可能起重要作用；②非萎缩组患者β-烯醇化酶mRNA的量有一定程度的升高,相应下游蛋白的表达与正常组比有升高趋势,表明DNA层面有一定的上调作用,同时翻译及其以后的修饰过程效率不高,可能受到某些抑制因素的调控,再者也有可能存在蛋白的过多消耗。 4.两种骨骼肌不同表型(萎缩型和非萎缩型)COPD,其下肢骨骼肌p-烯醇化酶的调控机制不同。
[Abstract]:Aim: to detect the expression of penolase protein and its mRNA in normal control group, COPD non-atrophied group and COPD atrophied group, and to explore the regulatory mechanism of penolase in two different phenotypes (atrophied type and non-atrophied type) COPD. Methods: the specimens of quadriceps femoris were collected in the first affiliated Hospital of Kunming Medical University, including 11 cases of normal control group, 14 cases of COPD non-atrophy group and 12 cases of COPD atrophy group. The levels of 尾-enolase mRNA and 尾-enolase protein were measured by RT-PCR in 37 patients and 37 patients with 尾-enolase protein by Western Blot. The mRNA transcription level and protein expression of p-enolase in quadriceps femoris were analyzed and compared among the three groups. The regulatory mechanism of 尾-enolase in two different phenotypes (atrophic type and non-atrophied type) COPD was investigated by combining the fatty index and the circumference of quadriceps femoris. Results: 1. The relative expression of 尾-enolase protein: 1 compared with the normal group, the expression of 尾-enolase protein in quadriceps femoris muscle of COPD skeletal muscle atrophy group was significantly higher than that of the normal group (P 0.05): 2 compared with the normal group, Although the expression of penolase protein in quadriceps femoris increased in COPD skeletal muscle non-atrophy group, the difference was not significant (P 0.05). 3 compared with COPD skeletal muscle atrophy group, there was no significant difference in the expression of 尾-enolase protein in quadriceps femoris between COPD skeletal muscle non-atrophy group and COPD skeletal muscle atrophy group (P 0.05). 2. The relative expression of p-enolase mRNA (2-螖 Ct): COPD skeletal muscle atrophy group, COPD skeletal muscle non-atrophy group, normal control group) had significant difference in the expression of 尾-enolase mRNA in quadriceps femoris (P 0.05). The expression of penolase in COPD skeletal muscle atrophy group was significantly higher than that in COPD skeletal muscle non-atrophy group, and the expression of p-enolase in COPD skeletal muscle non-atrophy group was higher than that in normal control group. The concentration of penolase protein and the expression of penolase mRNA in 3.COPD skeletal muscle atrophy group were significantly higher than those in COPD skeletal muscle non-atrophy group. The expression of penolase protein and penolase mRNA in skeletal muscle of COPD skeletal muscle non-atrophied group were increased. Conclusion: the concentration of 尾-enolase protein and the level of mRNA in quadriceps femoris in 1.COPD skeletal muscle atrophy group are higher than those in normal group, suggesting that the expression of penolase is up-regulated in COPD skeletal muscle atrophy patients, and there may be chronic hypoxia with COPD. The change of skeletal muscle type is related to the transformation of skeletal muscle type. The concentration of penolase protein and the level of penolase mRNA in 2.COPD skeletal muscle atrophy group were higher than those in COPD skeletal muscle non-atrophy group. There were differences in penolase protein concentration and penolase mRNA level between the two skeletal muscle phenotypes, indicating that different regulatory mechanisms may play different roles in the up-regulation of penolase by the two phenotypes. 3. There was a positive correlation between the concentration of penolase protein and the level of penolase nRNA in skeletal muscle of atrophied group and non-atrophied group: (1) the concentration of penolase protein was increased by increasing the amount of 尾-enolase mRNA in atrophied group. It is suggested that the regulation of DNA level may play an important role in the up-regulation of p-enolase. (2) the content of 尾-enolase mRNA in non-atrophied group increased to a certain extent, and the expression of downstream protein was higher than that in normal group, which indicated that DNA level was up-regulated to a certain extent, and the efficiency of translation and its subsequent modification process was not high. It may be regulated by some inhibitory factors, and there may also be excessive protein consumption. 4. The regulation mechanism of penolase in lower extremity skeletal muscle of two different phenotypes of skeletal muscle (atrophied and non-atrophied) COPD, is different.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R563.9
本文编号：2481271
[Abstract]:Aim: to detect the expression of penolase protein and its mRNA in normal control group, COPD non-atrophied group and COPD atrophied group, and to explore the regulatory mechanism of penolase in two different phenotypes (atrophied type and non-atrophied type) COPD. Methods: the specimens of quadriceps femoris were collected in the first affiliated Hospital of Kunming Medical University, including 11 cases of normal control group, 14 cases of COPD non-atrophy group and 12 cases of COPD atrophy group. The levels of 尾-enolase mRNA and 尾-enolase protein were measured by RT-PCR in 37 patients and 37 patients with 尾-enolase protein by Western Blot. The mRNA transcription level and protein expression of p-enolase in quadriceps femoris were analyzed and compared among the three groups. The regulatory mechanism of 尾-enolase in two different phenotypes (atrophic type and non-atrophied type) COPD was investigated by combining the fatty index and the circumference of quadriceps femoris. Results: 1. The relative expression of 尾-enolase protein: 1 compared with the normal group, the expression of 尾-enolase protein in quadriceps femoris muscle of COPD skeletal muscle atrophy group was significantly higher than that of the normal group (P 0.05): 2 compared with the normal group, Although the expression of penolase protein in quadriceps femoris increased in COPD skeletal muscle non-atrophy group, the difference was not significant (P 0.05). 3 compared with COPD skeletal muscle atrophy group, there was no significant difference in the expression of 尾-enolase protein in quadriceps femoris between COPD skeletal muscle non-atrophy group and COPD skeletal muscle atrophy group (P 0.05). 2. The relative expression of p-enolase mRNA (2-螖 Ct): COPD skeletal muscle atrophy group, COPD skeletal muscle non-atrophy group, normal control group) had significant difference in the expression of 尾-enolase mRNA in quadriceps femoris (P 0.05). The expression of penolase in COPD skeletal muscle atrophy group was significantly higher than that in COPD skeletal muscle non-atrophy group, and the expression of p-enolase in COPD skeletal muscle non-atrophy group was higher than that in normal control group. The concentration of penolase protein and the expression of penolase mRNA in 3.COPD skeletal muscle atrophy group were significantly higher than those in COPD skeletal muscle non-atrophy group. The expression of penolase protein and penolase mRNA in skeletal muscle of COPD skeletal muscle non-atrophied group were increased. Conclusion: the concentration of 尾-enolase protein and the level of mRNA in quadriceps femoris in 1.COPD skeletal muscle atrophy group are higher than those in normal group, suggesting that the expression of penolase is up-regulated in COPD skeletal muscle atrophy patients, and there may be chronic hypoxia with COPD. The change of skeletal muscle type is related to the transformation of skeletal muscle type. The concentration of penolase protein and the level of penolase mRNA in 2.COPD skeletal muscle atrophy group were higher than those in COPD skeletal muscle non-atrophy group. There were differences in penolase protein concentration and penolase mRNA level between the two skeletal muscle phenotypes, indicating that different regulatory mechanisms may play different roles in the up-regulation of penolase by the two phenotypes. 3. There was a positive correlation between the concentration of penolase protein and the level of penolase nRNA in skeletal muscle of atrophied group and non-atrophied group: (1) the concentration of penolase protein was increased by increasing the amount of 尾-enolase mRNA in atrophied group. It is suggested that the regulation of DNA level may play an important role in the up-regulation of p-enolase. (2) the content of 尾-enolase mRNA in non-atrophied group increased to a certain extent, and the expression of downstream protein was higher than that in normal group, which indicated that DNA level was up-regulated to a certain extent, and the efficiency of translation and its subsequent modification process was not high. It may be regulated by some inhibitory factors, and there may also be excessive protein consumption. 4. The regulation mechanism of penolase in lower extremity skeletal muscle of two different phenotypes of skeletal muscle (atrophied and non-atrophied) COPD, is different.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R563.9
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