早期乳酸林格氏液的復甦治療可以減輕大白鼠心臟受到創傷併發出血性休克的心臟凋亡; Early resuscitation by lactated Ringer’s solution alleviates cardiac apoptosis of trauma-hemorrhagic shock in rats

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Title:	早期乳酸林格氏液的復甦治療可以減輕大白鼠心臟受到創傷併發出血性休克的心臟凋亡;Early resuscitation by lactated Ringer’s solution alleviates cardiac apoptosis of trauma-hemorrhagic shock in rats
Authors:	劉建良;Chien-Liang Liu
Contributors:	中國醫藥大學：臨床醫學研究所碩士班
Keywords:	創傷;休克;心臟;凋亡;乳酸林格氏液;復甦;trauma;shock;heart;apoptosis;resuscitation
Date:	2009-07-15
Issue Date:	2009-08-12 14:23:33 (UTC+8)
Abstract:	背景：在因創傷而造成死亡中，創傷併發出血性休克往往是主要的原因。當身體在沒有胸腔或心臟傷害的機械性傷害的情況下，不論在臨床上或動物實驗中都發現有次發性的心臟傷害。凋亡在其中被認為扮演一個重要的角色。雖然乳酸林格氏液被認為可以減少創傷引起的發炎性物質及凋亡細胞的數目，但是鮮少研究發表針對心臟凋亡的效果。因此我們的動物實驗研究是否創傷併發出血性休克會引發心臟凋亡以及乳酸林格氏液復甦治療的效果，同時該治療效果的細胞內傳遞訊息路徑。研究方法：21 隻Sprague Dawley 雄性大鼠(300-350 克)被隨機分成對照組、出血休克組及休克後復甦組三組。創傷併發出血性模組藉由腹部切開後再縫合的傷口以及藉抽血造成的60 分鐘的休克而達成。復甦治療藉由乳酸林格氏液及抽出的血液在10 分鐘之內輸注，並恢復休克前血壓並維持穩定而完成。每一組大鼠在處置後在第0、4、8 小時作犧牲，藉由TUNEL 染色、HE 染色及西方染墨法將取出的心臟組織作研究。實驗結果：藉由HE 染色法發現休克組及復甦組的心肌細胞相對於對照組有明顯的減少。但是其中的復甦組減少的情形沒有休克組明顯。休克組的促凋亡信號( FasL, FADD, bax, bad) 均相對於對照組在有明顯的上升(p<0.05)。復甦治療組相對於休克組不但減少促凋亡信號外，同時可以增加磷酸化的IGF-1R,PI3K,AKt 訊號(p<0.05)。在TUNEL 染色方面，除偵測到休克組及復甦組中皆出現凋亡現象外，復甦組相對於休克組有明顯減少凋亡細胞的數目。結論：創傷併發出血性休克可以同時藉由fas/fasL 及粒線體依賴性的路徑引起心臟凋亡。但早期的乳酸林格氏液的復甦治療可以藉由減少促凋亡信號及活化IGF1R/PI3K/Akt 路徑來減輕心臟凋亡。 Background: Trauma complicated hemorrhagic shock is the leading cause in trauma deaths. Mechanical trauma of body without chest or cardiac injury would induce secondary cardiac damage in clinical and laboratory studies. Apoptosis was regarded as an important role. Although lactated Ringer’s (L/R)solution resuscitation would decrease the levels of cytokines and the amount of apoptotic cells, few reports discussed about the effects for cardiac apoptosis. Therefore we investigated whether trauma-hemorrhagic shock (THS) would induce cardiac apoptosis, in addition, the therapeutic effects and the intracellular mechanism of L/R solution resuscitation. Material and Method: Twenty-one male Sprague Dawley rats (300~350 g) were randomly divided into sham (C),hemorrhagic shock (S), and hemorrhagic shock with resuscitation (R) groups.THS was induced by the cutting wound sutured over abdomen and maintained 60 minutes shock by withdrawal blood. Then resuscitation with auto-transfusion and L/R in 10 minutes after shock restored pre-shock blood pressure. Each group was scarified in different hours after intervention at zero hour, four hours, and eight hours. Cardiac tissue were collected and evaluated for apoptosis by TdT-mediated dUTP nick-end labeling (TUNEL) stain, Haematoxylin & Eosin(HE) stain, and western blot. Result: HE stain showed significant decrease of cardiomyocyte amount compared S group to C group and R group. But Less decrease of that in R group compared to S group. S group had significant elevation of pro-apoptotic signals (fas/fasL, fas associated death domain (FADD), bax, bad) compared to C group (p< 0.05). Resuscitation can decrease the pro-apoptotic signals, in addition, increased of phophorated IGF-1R,phosphorated PI3K, and Akt amounts compared between R and S group (p<0.05). TUNEL stain showed existence of apoptotic cells in both S and R groups. R group significant decrease the cell amount of apoptosis compared to S group. Conclusion: THS induces cardiac apoptosis by fas/fasL and mitochondrial dependent pathways, and early L/R solution resuscitation can alleviate cardiac apoptosis by decrease the amounts of pro-apoptotic signals and activate IGF1R/PI3K/Akt pathway.
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