β-烯醇化酶及mRNA在COPD患者下肢骨骼肌表達水平研究
發(fā)布時間：2019-05-20 02:29
【摘要】：目的：測定p-烯醇化酶蛋白及其mRNA水平在正常對照組、COPD非萎縮組、COPD萎縮組表達情況,探討p-烯醇化酶在兩種不同表型(萎縮型和非萎縮型)COPD的調(diào)控機制。 方法：收集昆明醫(yī)科大學(xué)第一附屬醫(yī)院骨科手術(shù)診療患者股四頭肌標(biāo)本,其中正常對照組11例,COPD非萎縮組14例,COPD萎縮組12例。采用RT-PCR測定37例患者β-烯醇化酶mRNA水平,采用Western Blot技術(shù)檢測37例患者β-烯醇化酶蛋白,分析比較3組患者股四頭肌p-烯醇化酶mRNA轉(zhuǎn)錄水平及蛋白表達差異,結(jié)合患者去脂肪指數(shù)及股四頭肌周徑等影響因素,探討β-烯醇化酶在兩種不同表型(萎縮型和非萎縮型)COPD的調(diào)控機制。 結(jié)果：1.β-烯醇化酶蛋白相對表達量：①與正常組比較,COPD骨骼肌萎縮組患者股四頭肌中β-烯醇化酶蛋白表達顯著升高(P0.05)：②與正常組比較,COPD骨骼肌非萎縮組患者股四頭肌p-烯醇化酶蛋白表達雖然有升高趨勢,但差異不顯著(P0.05)；③與COPD骨骼肌萎縮組比較,COPD骨骼肌非萎縮組患者股四頭肌β-烯醇化酶蛋白表達無差異(P0.05)。 2.p-烯醇化酶mRNA的相對表達量(2-ΔΔCt):COPD骨骼肌萎縮組、COPD骨骼肌非萎縮組、正常對照組患者股四頭肌β-烯醇化酶mRNA表達具有差異性(P0.05),進一步兩兩比較,各組間差異均有統(tǒng)計學(xué)意義,COPD骨骼肌萎縮組中p-烯醇化酶表達明顯高于COPD骨骼肌非萎縮組、正常對照組,COPD骨骼肌非萎縮組p-烯醇化酶表達高于正常對照組。 3.COPD骨骼肌萎縮組p-烯醇化酶蛋白濃度與p-烯醇化酶mRNA表達水平一致升高,COPD骨骼肌非萎縮組骨骼肌中p-烯醇化酶蛋白濃度與p-烯醇化酶mRNA表達水平呈升高趨勢。 結(jié)論：1.COPD骨骼肌萎縮組患者股四頭肌β-烯醇化酶蛋白濃度及mRNA水平較正常組高,提示COPD骨骼肌萎縮患者p-烯醇化酶表達上調(diào),可能與COPD存在慢性缺氧、骨骼肌類型轉(zhuǎn)換有關(guān)； 2.COPD骨骼肌萎縮組患者p-烯醇化酶蛋白濃度及p-烯醇化酶mRNA水平較COPD骨骼肌非萎縮組高,p-烯醇化酶蛋白濃度及p-烯醇化酶mRNA水平在兩種骨骼肌表型之間存在差異,表明可能不同的調(diào)控機制在兩種表型對p-烯醇化酶的上調(diào)發(fā)揮了不同作用； 3.萎縮組及非萎縮組骨骼肌p-烯醇化酶蛋白濃度與p-烯醇化酶nRNA水平有一定的正相關(guān)性：①萎縮組患者通過增加β-烯醇化酶mRNA的量使p-烯醇化酶蛋白增高,說明DNA層面的調(diào)控在p-烯醇化酶的上調(diào)中可能起重要作用；②非萎縮組患者β-烯醇化酶mRNA的量有一定程度的升高,相應(yīng)下游蛋白的表達與正常組比有升高趨勢,表明DNA層面有一定的上調(diào)作用,同時翻譯及其以后的修飾過程效率不高,可能受到某些抑制因素的調(diào)控,再者也有可能存在蛋白的過多消耗。 4.兩種骨骼肌不同表型(萎縮型和非萎縮型)COPD,其下肢骨骼肌p-烯醇化酶的調(diào)控機制不同。
[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.
【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R563.9
本文編號：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.
【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R563.9
【共引文獻】
相關(guān)期刊論文 前7條
1 符曉蘇;張志宇;孫奕;;中樞神經(jīng)損傷疾病與烯醇化酶活性的相關(guān)性探討[J];中國老年保健醫(yī)學(xué);2011年06期
2 李天伯,胡洋,王以光,夏煥章;mMR-1基因的克隆和在畢赤酵母中分泌表達[J];生物工程學(xué)報;2005年01期
3 朱理安;方寧遠;;α烯醇化酶——古老的蛋白,嶄新的功能[J];國際病理科學(xué)與臨床雜志;2007年04期
4 黃建珍;唐雪;阮記明;馬海田;鄒思湘;;肉種雞飼養(yǎng)方式對子代胚肝蛋白表達譜的影響[J];畜牧獸醫(yī)學(xué)報;2009年06期
5 陳鴻軍;沈欣悅;陳丹清;于圣青;丁鏟;;雞毒支原體烯醇化酶單克隆抗體研制及黏附阻斷[J];中國獸醫(yī)學(xué)報;2012年06期
6 唐江偉;;腦梗死神經(jīng)元特異性烯醇化酶的變化及其臨床意義[J];中國現(xiàn)代藥物應(yīng)用;2012年13期
7 王庭利;劉百歌;王玲玲;趙海豐;何成彥;劉寧;郭宏華;;α-enolase在大腸癌與癌旁組織中的表達及意義[J];中國實驗診斷學(xué);2013年06期
相關(guān)博士學(xué)位論文 前4條
1 姚欣;腹瀉型腸易激綜合征患者結(jié)腸黏膜蛋白質(zhì)的篩選與鑒定[D];中國人民解放軍軍醫(yī)進修學(xué)院;2008年
2 黃建珍;胚胎期肉雞肝臟脂類代謝關(guān)鍵因子的篩選及DHEA的調(diào)控研究[D];南京農(nóng)業(yè)大學(xué);2008年
3 孟憲宇;周圍神經(jīng)感覺束和運動束的比較蛋白質(zhì)組學(xué)研究[D];吉林大學(xué);2010年
4 趙海豐;Dukes B期大腸癌蛋白質(zhì)組學(xué)研究[D];吉林大學(xué);2010年
相關(guān)碩士學(xué)位論文 前4條
1 王仕欽;白念珠菌烯醇化酶及其N端肽段的原核表達和應(yīng)用研究[D];南京師范大學(xué);2011年
2 郭遠盤;8周有氧運動對大鼠左心室肌蛋白質(zhì)組差異表達的影響[D];湖南師范大學(xué);2010年
3 沈欣悅;雞毒支原體兩種膜相關(guān)酶的生物學(xué)功能研究[D];揚州大學(xué);2010年
4 宿魯鋒;表沒食子兒茶素沒食子酸酯、β-羥基-β甲基丁酸鹽聯(lián)合治療癌癥惡病質(zhì)實驗性研究[D];福建醫(yī)科大學(xué);2013年
，本文編號：2481271
本文鏈接：http://sikaile.net/yixuelunwen/huxijib/2481271.html
最近更新
教材專著
