【摘要】：目的：研究慢性阻塞性肺疾病模型大鼠骨骼肌以及血清中IGF-1的表達、骨骼肌細胞中細胞凋亡相關(guān)基因Bcl-2,Bax, Caspase-3的表達,并探討是否由于COPDD大鼠體內(nèi)IGF-1表達降低,影響B(tài)cl-2/Bax的比值,從而激活Caspase-3蛋白,產(chǎn)生細胞凋亡級聯(lián)反應(yīng),最終導(dǎo)致骨骼肌萎縮。 方法：70只健康Wistar大鼠,分為COPD模型35只,正常對照組35只。其中COPD模型組在第1-14天、第16-104天放入自制的染毒箱內(nèi)熏煙,第15天在氣管內(nèi)注射胰蛋白酶(PPE)造COPD大鼠模型。正常對照組大鼠被飼養(yǎng)在無香煙煙霧污染的飼養(yǎng)房內(nèi),除了第15天在氣管內(nèi)注射生理鹽水外,飼養(yǎng)期間不做任何處理。兩組大鼠同時飼養(yǎng)至第104天。第104天取大鼠的肺組織以及趾長伸肌,其中一側(cè)趾長伸肌制成石蠟切片,另一側(cè)趾長伸肌放入-80℃冰箱中；取大鼠動脈血提取血清；測量各組大鼠的體重以及趾長伸肌的重量。采用HE染色法觀察兩組大鼠肺組織結(jié)構(gòu)的改變,免疫組織化學法測定趾長伸肌的IGF-1,Bcl-2,Bax,Caspase-3的表達。并采用ELISA(酶聯(lián)免疫分析法)測定血清中IGF-1的表達。用Image-Pro Plus Version6.0圖像處理系統(tǒng)測定免疫組化強度,Werstern-Blot法測定趾長伸肌Caspase-3活性蛋白表達。所得數(shù)據(jù)經(jīng)SPSS17.0軟件做單因素的方差分析、t檢驗以及相關(guān)性分析。 結(jié)果： 1.COPD模型組大鼠的氣管壁以及肺組織均有大量炎癥細胞浸潤,通過進行大鼠肺功能測量,以及觀察肺組織的病理切片,判定COPD模型組大鼠肺符合COPD診斷標準。 2.COPD模型組大鼠體重以及一側(cè)趾長伸肌的重量較正常對照組均有明顯減輕(P0.05,P0.05)。 3.COPD模型組大鼠血清以及趾長伸肌中IGF-1表達明顯低于正常對照組大鼠(P0.05,P0.05)。 4.正常對照組大鼠趾長伸肌中Bcl-2的表達明顯強于COPD模型組大鼠(P0.05,P0.05) 5. COPD模型組大鼠趾長伸肌中Bax以及Caspase-3的表達明顯強于對照組大鼠(P0.05,P0.05) 6.各組大鼠血清以及趾長伸肌IGF-1的表達與Bcl-2/Bax的表達相關(guān)性分析呈正相關(guān)(P0.05) 7.各組大鼠血清以及趾長伸肌肉IGF-1的表達與Caspse-3的表達相關(guān)性分析呈負相關(guān)(P0.05)。 結(jié)論： 1.COPD模型血清以及骨骼肌IGF-1表達均明顯減少。 2.骨骼肌細胞凋亡率增加可能是COPD模型大鼠骨骼肌萎縮的重要機制。 3.COPD大鼠體內(nèi)IGF-1在體內(nèi)表達減少可能通過降低Bcl-2/Bax的比值,從而激活Caspase-3活性蛋白,產(chǎn)生細胞凋亡級聯(lián)反應(yīng),最終導(dǎo)致骨骼肌萎縮。
[Abstract]:Objective: to study the expression of IGF-1 in skeletal muscle and serum and the expression of apoptosis-related gene Bcl-2,Bax, Caspase-3 in skeletal muscle cells of rats with chronic obstructive pulmonary disease (COPD). The purpose of this study was to explore whether the decrease of IGF-1 expression in COPDD rats could affect the ratio of Bcl-2/Bax, activate Caspase-3 protein, produce apoptosis cascade reaction, and finally lead to skeletal muscle atrophy. Methods: 70 healthy Wistar rats were divided into COPD model (35 rats) and normal control group (35 rats). In the COPD model group, COPD rats were induced by intratracheal injection of trypsin (PPE) on the 1st-14th day, 16th-104th day and 16-104th day. The rats in the normal control group were fed into a room without cigarette smoke pollution. No treatment was given during the feeding period except for the injection of saline into the trachea on the 15th day. The rats in both groups were fed at the same time until the 104th day. On the 104th day, the lung tissue and extensor digitorum longus muscle of rats were taken, in which one extensor digitorum longus muscle was made into paraffin section, the other side extensor digitorum longus muscle was put into refrigerator at-80 鈩，

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