大鼠骨骼肌缺血后調(diào)適動態(tài)pH測量及其模擬酸灌注減輕缺血再灌注損傷研究
發(fā)布時間：2018-07-29 15:34
【摘要】：背景:近來有研究報道,心肌缺血后調(diào)適保護(hù)的重要機(jī)制是通過心肌缺血后再灌注初期維持3min的酸中毒狀態(tài)起到作用。但其具體的p H變化如何以及模擬酸性灌注能否減輕缺血再灌注損傷未見深入研究。在骨骼肌方面的類似研究更亦未查見。目的:對大鼠骨骼肌缺血后調(diào)適過程進(jìn)行動態(tài)p H測量,并模擬缺血后調(diào)適p H變化配置酸性灌注液輸注大鼠體內(nèi),檢測其對缺血再灌注損傷的影響。方法:基于前期大鼠骨骼肌缺血再灌注損傷建模方法和缺血后調(diào)適優(yōu)化方案,運用光纖p H儀于主缺血期、4個循環(huán)30s再灌/30s缺血后調(diào)適操作期及其后再灌注期對大鼠骨骼肌進(jìn)行p H連續(xù)測量,并用乳酸及生理鹽水配置與缺血后調(diào)適期等p H灌注液備用。將25只健康成年雄性SD大鼠隨機(jī)分為假手術(shù)組、缺血再灌注組、缺血后調(diào)適組、乳酸酸灌注組、生理鹽水組5組,每組5只。各組按其實驗方案進(jìn)行相應(yīng)實驗并抽血檢測乳酸脫氫酶,取樣腓腸肌測算濕/干質(zhì)量比值、髓過氧化物酶及2,3,5—氯化三苯基四氮唑(TTC)染色測算梗死面積,取樣右側(cè)脛前肌行Western Blot檢測MAPK通路關(guān)鍵蛋白Erk1/2的表達(dá)。結(jié)果:(1)缺血后調(diào)適于再灌注初期出現(xiàn)一個延長酸性平臺,p H為6.81±0.133,時長為2min40s。(2)乳酸脫氫酶、髓過氧化物酶、濕/干質(zhì)量比值檢測結(jié)果顯示:缺血后調(diào)適組、乳酸輸注組均明顯低于缺血再灌注組(P0.05);(3)Western Blot分析顯示:p-Erk的表達(dá)缺血后調(diào)適組、乳酸輸注組及生理鹽水輸注組均顯著高于缺血再灌注組(P0.05)。(4)TTC染色檢測顯示,缺血后調(diào)適組及乳酸輸注組梗死面積較缺血再灌注組明顯減少(P0.05)。結(jié)論:缺血后調(diào)適可維持再灌注初期短暫酸性狀態(tài)。酸性灌注液可以有效模擬缺血后調(diào)適作用,減輕大鼠骨骼肌缺血再灌注損傷。其機(jī)制可能直接作用于MPTP避免其即時開放,或通過Erk1/2磷酸化激活RISK信號通路而起到保護(hù)作用。
[Abstract]:Background: recently, it has been reported that the important mechanism of myocardial ischemic conditioning is to maintain the acidosis state of 3min in the early stage of myocardial ischemia and reperfusion. However, the specific pH changes and whether simulated acid perfusion can reduce ischemia reperfusion injury have not been further studied. Similar studies on skeletal muscle have also not been found. Aim: to measure the dynamic pH of skeletal muscle after ischemia in rats, and to investigate the effects of pH on ischemia reperfusion injury in rats. Methods: based on the modeling method of ischemic reperfusion injury of skeletal muscle in early stage and the optimized scheme of adjustment after ischemia in rat skeletal muscle. The pH of skeletal muscle of rats was continuously measured by fiber-optic pH instrument during the main ischemia period, 4 cycles 30 s / 30 s ischemia conditioning operation period and subsequent reperfusion period. The pH perfusion solution was prepared with lactic acid and physiological saline configuration and after ischemia adaptation period. Twenty-five adult male SD rats were randomly divided into sham-operation group, ischemia-reperfusion group, post-ischemic conditioning group, lactic acid perfusion group, physiological saline group, 5 rats in each group. According to the experimental plan, each group was tested for lactate dehydrogenase, the wet / dry weight ratio of gastrocnemius was measured by sampling gastrocnemius muscle, and the infarct area was measured by myeloperoxidase and (TTC) staining. The expression of Erk1/2, a key protein in the MAPK pathway, was detected by Western Blot in the right tibial anterior muscle. Results: (1) the pH of a prolonged acidic platform was 6.81 鹵0.133, and the duration was 2 min 40 s. (2) lactate dehydrogenase, myeloperoxidase and wet / dry weight ratio were measured. The expression of p-Erk in the lactic acid infusion group was significantly lower than that in the ischemia reperfusion group (P0.05); (3) Western Blot analysis showed that the expression of p-Erk was significantly higher in the lactic acid infusion group and saline infusion group than in the ischemia reperfusion group (P0.05). (4) TTC staining. The infarct size of conditioning group and lactic acid infusion group was significantly lower than that of ischemia reperfusion group (P0.05). Conclusion: conditioning after ischemia can maintain the transient acidic state in the early stage of reperfusion. Acid perfusion solution can effectively simulate the effects of ischemic conditioning and alleviate the ischemia reperfusion injury of skeletal muscle in rats. The mechanism may play a direct role in MPTP to avoid its immediate opening, or through Erk1/2 phosphorylation to activate the RISK signaling pathway and play a protective role.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R68
本文編號：2153140
[Abstract]:Background: recently, it has been reported that the important mechanism of myocardial ischemic conditioning is to maintain the acidosis state of 3min in the early stage of myocardial ischemia and reperfusion. However, the specific pH changes and whether simulated acid perfusion can reduce ischemia reperfusion injury have not been further studied. Similar studies on skeletal muscle have also not been found. Aim: to measure the dynamic pH of skeletal muscle after ischemia in rats, and to investigate the effects of pH on ischemia reperfusion injury in rats. Methods: based on the modeling method of ischemic reperfusion injury of skeletal muscle in early stage and the optimized scheme of adjustment after ischemia in rat skeletal muscle. The pH of skeletal muscle of rats was continuously measured by fiber-optic pH instrument during the main ischemia period, 4 cycles 30 s / 30 s ischemia conditioning operation period and subsequent reperfusion period. The pH perfusion solution was prepared with lactic acid and physiological saline configuration and after ischemia adaptation period. Twenty-five adult male SD rats were randomly divided into sham-operation group, ischemia-reperfusion group, post-ischemic conditioning group, lactic acid perfusion group, physiological saline group, 5 rats in each group. According to the experimental plan, each group was tested for lactate dehydrogenase, the wet / dry weight ratio of gastrocnemius was measured by sampling gastrocnemius muscle, and the infarct area was measured by myeloperoxidase and (TTC) staining. The expression of Erk1/2, a key protein in the MAPK pathway, was detected by Western Blot in the right tibial anterior muscle. Results: (1) the pH of a prolonged acidic platform was 6.81 鹵0.133, and the duration was 2 min 40 s. (2) lactate dehydrogenase, myeloperoxidase and wet / dry weight ratio were measured. The expression of p-Erk in the lactic acid infusion group was significantly lower than that in the ischemia reperfusion group (P0.05); (3) Western Blot analysis showed that the expression of p-Erk was significantly higher in the lactic acid infusion group and saline infusion group than in the ischemia reperfusion group (P0.05). (4) TTC staining. The infarct size of conditioning group and lactic acid infusion group was significantly lower than that of ischemia reperfusion group (P0.05). Conclusion: conditioning after ischemia can maintain the transient acidic state in the early stage of reperfusion. Acid perfusion solution can effectively simulate the effects of ischemic conditioning and alleviate the ischemia reperfusion injury of skeletal muscle in rats. The mechanism may play a direct role in MPTP to avoid its immediate opening, or through Erk1/2 phosphorylation to activate the RISK signaling pathway and play a protective role.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R68
【參考文獻(xiàn)】
相關(guān)期刊論文 前5條
1 賈彤;王會娟;王麗;邢珍;李福龍;;右美托咪定對老年患者體外循環(huán)心肌缺血再灌注損傷的保護(hù)作用[J];中國臨床藥理學(xué)雜志;2016年16期
2 Wang-sheng Duanmu;Liu Cao;Jing-yu Chen;Hong-fei Ge;Rong Hu;Hua Feng;;Ischemic postconditioning protects against ischemic brain injury by up-regulation of acid-sensing ion channel 2a[J];Neural Regeneration Research;2016年04期
3 彭龍龍;陽富春;薄占東;譚楨;姚軍;程建文;薛明強(qiáng);趙勁民;;不同缺血后調(diào)適方案對大鼠模型肢體骨骼肌缺血再灌注損傷的影響[J];中國組織工程研究;2015年05期
4 喻琴琴;楊俊;郭鑫;;心肌缺血再灌注損傷的天然免疫學(xué)機(jī)制進(jìn)展[J];實用醫(yī)學(xué)雜志;2014年15期
5 魏育濤;杜俊峰;朱佳龍;羅波;;局部缺血預(yù)適應(yīng)對兔短期缺血再灌注心肌細(xì)胞凋亡及Bcl-2和Bax蛋白表達(dá)的影響[J];中國組織工程研究與臨床康復(fù);2007年21期
，本文編號：2153140
本文鏈接：http://sikaile.net/shoufeilunwen/mpalunwen/2153140.html
最近更新
教材專著
