【摘要】：目的：近年過度訓(xùn)練逐漸成為運(yùn)動(dòng)醫(yī)學(xué)、臨床醫(yī)學(xué)的研究熱點(diǎn)。過度訓(xùn)練致急性心肌損傷（overtraining-induced acute myocardial injury，OTIAMI）是運(yùn)動(dòng)員發(fā)生猝死的重要原因之一，目前尚缺乏有效的防治措施，因此對(duì)OTIAMI機(jī)制及防治措施的研究具有重要的臨床意義。本研究采用游泳力竭法建立過度訓(xùn)練致大鼠急性心肌損傷模型，探討細(xì)胞色素C及Caspase-3在過度訓(xùn)練致急性心肌損傷中的作用及旋覆花素的干預(yù)效應(yīng)，從而為OTIAMI的防治提供新的實(shí)驗(yàn)依據(jù)和治療靶點(diǎn)。 方法：健康雄性Wistar大鼠48只，體重200～220g，采用隨機(jī)數(shù)字表法，將大鼠隨機(jī)分為3組：安靜對(duì)照組（control，C組，n=8）、力竭運(yùn)動(dòng)組（exhaustive swimming，ES組，n=24）、旋覆花素干預(yù)組（InulaBritannica，IB組，n=16）。C組不做任何處理；ES組及IB組采用游泳力竭法建立大鼠過度訓(xùn)練急性心肌損傷模型，IB組于力竭前24h及力竭前即刻分別口腔灌入旋覆花素25ml/kg后進(jìn)行游泳直至力竭。各組大鼠于實(shí)驗(yàn)前一天適應(yīng)性游泳30min，次日再次放于水中游泳直至力竭。力竭標(biāo)準(zhǔn)：①沉入水底超過10s；②游泳動(dòng)作明顯不協(xié)調(diào)；③撈出后沒有逃避反應(yīng)，且放在平面上無法完成翻正反射。 ES組于力竭后即刻（ESI）、力竭后6h（ES6h）和力竭后24h（ES24h）；IB組于力竭后6h（IB6h）、力竭后24h（IB24h）分別隨機(jī)抽取8只大鼠，乙醚麻醉下采集下腔靜脈血3ml，，3000轉(zhuǎn)/min，離心15min，取上清液，-20℃保存，采血后摘取心臟，取心尖同一部位心肌組織置于4%多聚甲醛固定。光鏡下觀察大鼠心肌組織病理學(xué)變化，采用末端脫氧核苷酸轉(zhuǎn)移酶介導(dǎo)的缺口末端標(biāo)記法（TUNEL)觀察心肌細(xì)胞凋亡情況，ELISA法檢測(cè)血清心肌肌鈣蛋白I（cTnI）濃度，免疫組化法檢測(cè)心肌組織細(xì)胞色素C及Caspase-3表達(dá)變化。 統(tǒng)計(jì)學(xué)處理采用SPSS13.0統(tǒng)計(jì)學(xué)軟件進(jìn)行分析，計(jì)量資料以均數(shù)±標(biāo)準(zhǔn)差（x±s）表示，組間比較以及ES組各時(shí)間點(diǎn)的比較采用單因素方差分析，P＜0.05為差異有統(tǒng)計(jì)學(xué)意義。 結(jié)果：1.心肌組織病理學(xué)變化C組大鼠心肌細(xì)胞呈橢圓形或短梭形，心肌纖維互相連接排列緊密，可見清晰的橫紋。ES組大鼠力竭即刻見部分心肌纖維腫脹肥大，偶見心肌間質(zhì)血管及毛細(xì)血管擴(kuò)張；力竭后6h時(shí)可見部分心肌細(xì)胞的細(xì)胞核深染，有固縮現(xiàn)象，心肌纖維腫脹肥大明顯，排列紊亂，出現(xiàn)不規(guī)整斷裂；力竭后24h時(shí)心肌纖維腫脹肥大有所好轉(zhuǎn)，心肌細(xì)胞間散在大量炎癥浸潤灶。IB組各時(shí)點(diǎn)心肌組織病理學(xué)變化明顯減輕。 2.各組大鼠不同時(shí)點(diǎn)血清cTnI的比較與C組比較，ES組及IB組過度訓(xùn)練后各時(shí)點(diǎn)大鼠血清cTnI濃度升高（P＜0.05）；與ES組比較，IB組過度訓(xùn)練后6h、24h時(shí)大鼠血清cTnI濃度下降（P＜0.05）；與力竭后即刻相比，力竭后6h時(shí)大鼠血清cTnI濃度明顯升高（P 0.05），24h時(shí)大鼠血清cTnI濃度有所降低(P＜0.05）。 3.各組大鼠不同時(shí)點(diǎn)心肌細(xì)胞凋亡指數(shù)的比較與C組比較，ES組及IB組力竭后各時(shí)點(diǎn)大鼠心肌細(xì)胞凋亡指數(shù)增加（P＜0.05）；與ES組比較，IB組力竭后6h及24h時(shí)大鼠心肌細(xì)胞凋亡指數(shù)明顯降低（P＜0.05）；與力竭后即刻比較，力竭后6h及24h時(shí)大鼠心肌細(xì)胞凋亡指數(shù)增加（P＜0.05），且力竭后6h時(shí)達(dá)高峰，24h時(shí)有所下降（P＜0.05）。 4.各組大鼠心肌組織不同時(shí)點(diǎn)細(xì)胞色素C及Caspase-3表達(dá)的比較各組大鼠心肌組織各時(shí)間點(diǎn)Caspase-3與細(xì)胞色素C變化趨勢(shì)一致。與C組比較，ES組及IB組力竭后各時(shí)點(diǎn)大鼠心肌組織細(xì)胞色素C及Caspase-3表達(dá)上調(diào)（P＜0.05）；與ES組比較，IB組力竭后6h、24h時(shí)大鼠心肌組織細(xì)胞色素C及Caspase-3表達(dá)下調(diào)（P＜0.05）；ES組力竭后6h、24h時(shí)與力竭后即刻比較，大鼠心肌組織細(xì)胞色素C及Caspase-3表達(dá)上調(diào)（P＜0.05），且力竭后6h時(shí)表達(dá)最強(qiáng)，24h時(shí)有所下調(diào)（P＜0.05）。 結(jié)論：1.游泳所致力竭可引起心肌損傷，但病變較輕，受損心肌可在24h內(nèi)部分修復(fù)。 2.游泳致力竭運(yùn)動(dòng)可引起大鼠心肌細(xì)胞凋亡，心肌細(xì)胞凋亡可能為過度訓(xùn)練致大鼠心肌損傷的病理生理學(xué)基礎(chǔ)之一。 3.細(xì)胞色素C及Caspase-3通過線粒體信號(hào)轉(zhuǎn)導(dǎo)通路介導(dǎo)了過度訓(xùn)練致大鼠急性心肌損傷時(shí)心肌細(xì)胞凋亡。 4.旋覆花素可減輕過度訓(xùn)練致大鼠急性心肌損傷，其機(jī)制可能與其抑制心肌細(xì)胞凋亡相關(guān)信號(hào)轉(zhuǎn)導(dǎo)因子Caspase-3及細(xì)胞色素C表達(dá)，減少心肌細(xì)胞凋亡有關(guān)，及時(shí)采取藥物干預(yù)可能為OTIAMI防治的關(guān)鍵。
[Abstract]:Objective: Over-training has become a hot topic in sports medicine and clinical medicine in recent years. Overtraining induced acute myocardial injury (OTIAMI) is one of the most important causes of sudden death in athletes. The effects of cytochrome C and Caspase-3 on acute myocardial injury induced by over-training induced by over-training were studied by means of swimming exhaustion method. The effect of cytochrome C and Caspase-3 on acute myocardial injury was studied. Methods: Forty-eight healthy male Wistar rats were randomly divided into three groups: control group (control group, group C, n = 8), exhaustive exercise group (ES group, n = 24). 6) No treatment was done in group C; ES group and IB group used swimming exhaustion method to establish the model of acute myocardial injury in rats. After exhaustive swimming, the rats were allowed to swim for 30minutes the day before the experiment, and then put it again in water the next day until they were allowed to swim. Exhaustion. Exhaustive standard: the water is sunk to the water bottom for more than 10s; the swimming action of the swimming pool is obviously not coordinated; after fishing, no escaping reaction is avoided, and the turning is not completed on the plane. The results were as follows: after exhaustive exhaustion (ESI), 6h (ES6h) and 24h (ES24h) after exhaustive exhaustion, 8 rats were randomly sampled from the group IB after exhaustive exhaustion (IB6h). After exhaustive exhaustion, 8 rats were sampled at random, 3ml of venous blood were collected under ether anesthesia, then centrifuged for 15min, supernatant was removed, and blood was collected at -20 鈩

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