本文關(guān)鍵詞：急進高原環(huán)境對大鼠心肌損傷作用及藥物保護機制研究　出處：《蘭州大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 低氧低氣壓 高原急性缺氧 心肌標志物 氧化應(yīng)激 美托洛爾 線粒體 呼吸鏈

【摘要】：目的 1.以55m、1500m、2260m、3400m、4300m五個海拔進行梯度實驗,研究大鼠急進高原環(huán)境后心肌的受損情況及趨勢,探討氧化應(yīng)激與線粒體在心肌損傷中的作用； 2.以氧化應(yīng)激、生化、血氣、形態(tài)顯微觀察等指標綜合評價四種藥物干預(yù)后,對急進高原大鼠心肌的影響,篩選出效果優(yōu)良的藥物； 3.探索藥物美托洛爾保護急進高原大鼠心肌過氧化損傷與線粒體損傷的機制。 方法 實驗一：大鼠隨機分為5組每組12只,A組(上海55m)、B組(甘肅蘭州1500m)、C組(青海西寧2260m)、D組(甘肅碌曲3400m)、E組(青�，敹�4300m),記錄體重后分別從平原地區(qū)(上海55m)急進實地海拔各組處。3d后取靜脈血清測定生化全項,動脈血測血氣等指標,心肌組織固定做病理、電鏡觀察及液氮冷凍做氧化水平測定；實驗二：急進3400m后分別給予四種藥物：胺腆酮、美托洛爾、卡托普利、呋噻米及陽性對照藥物地塞米松。記錄1d及3d體重,3d后給藥組大鼠的心率、血壓,并采眼眶靜脈血,用于生化項目檢測(血清CK、CK-MB、AST、HDBH、LDH等)及心肌標志物測定(MYO、Tn-I);所有實驗動物麻醉后腹主動脈采集動脈血進行血氣分析(ph、pO2、pCO2、Hct、ctHb、 cHCO3、BB、 BE、SO2等)；活體取大鼠心臟組織記錄重量,切取部分固定進行病理及電鏡觀察；剩余心肌組織用液氮進行快速冷凍保存,用于測定氧化水平(SOD、MDA、NO);實驗三：平原組、海拔3400m空白組及美托洛爾組取心臟勻漿進行線粒體復(fù)合體(COM-Ⅰ、 COM-Ⅱ、COM-Ⅲ、COM-Ⅳ)活性,MDH. SDH及ATP活性測定。 結(jié)果 1.實驗一各組大鼠心肌酶譜結(jié)果B、C、D、E組大鼠較A組明顯增加,其中CK及CK-MB在B組分別增加了43.87%、55.36%,C組增加了185.14%、84.54%,D組增加了73.54%、139.46%,E組增加了118.28%、175.32%(P0.05)。因此B組(1500m)、C組(2260m)、D組(3400m)、E組(4300m)心肌酶譜在血液中的釋放明顯增加;各組大鼠心肌標志物結(jié)果：與A組相比MYO、Tn-I在C、D、E組均顯著增加,其中C組增加了182.82%、21.30%,D組增加了328.35%、54.73%,E組增加了444.04%、62.67%(P0.05),標志物結(jié)果顯示了在C組(2260m)、D組(3400m)、E組(4300m)的心肌損傷較平原組明顯增加； 2.實驗一與A組相比各組大鼠氧化應(yīng)激指標顯示,SOD在B、C、D、E組分別減少10.06%、13.51%、12.18%、29.92%；MDA在B、C、D、E組分別增加14.67%、89.33%、36.01%、86.66%；NO在B、C、D、E組分別減少64.45%、65.49%、66.94%、40.54%(P0.05),提示在C、D、E組大鼠心肌氧化應(yīng)激水平增加,動脈氧分壓降低明顯； 3.實驗一大鼠動脈氧分壓及二氧化碳分壓明顯降低(P0.05)(海拔≥2260m),與A組相比,pCO2在B、C、D、E分別下降9.56%、6.68%、25.07%、32.01%；pO2在B、C、D、E分別下降7.95%、11.97%、48.94%、43.09%；s02在D、E組明顯下降分別下降12.04%、8.25%(P0.05); 4.實驗二,在3400m海拔處應(yīng)用藥物后對過氧化損傷均有不同改善作用,其中美托洛爾組SOD及NO較高原空白組分別增加10.75%、144.65%(P0.05),MDA減少25.49%(P0.05),并且提高氧分壓使之增加31.68%(P0.05),能明顯降低心肌酶譜增加的情況； 5.實驗三,在3400m海拔處線粒體復(fù)合體Ⅰ、Ⅱ、Ⅲ、Ⅳ的活性在高原模型組較平原組明顯下降,其中COM-Ⅰ、COM-Ⅱ、COM-III下降顯著,與平原組相比在高原地區(qū)COM-Ⅰ下降了26.17%,COM-Ⅱ下降了24.21%,COM-Ⅲ下降了17.94%,COM-IV下降了32.61%(P0.05),應(yīng)用藥物美托洛爾后明顯提高其活力,分別較高原組上升22.48%、44.01%、36.01%、17.23%(P0.05),同樣ATP各含量在應(yīng)用藥物后明顯增加(P0.05)； 6.實驗一及實驗二的心肌組織病理及電鏡結(jié)果顯示,海拔≥2260m損傷較平原組顯著,呈現(xiàn)肌纖維的不整齊排列,線粒體腫脹溶解,閏盤加寬等,給予藥物后有不同程度的改善,其中美托洛爾保護作用較明顯。 結(jié)論 1.實驗動物從平原55m急進1500m、2260m、3400m、4300m不同海拔后,氧化應(yīng)激結(jié)果顯示高原環(huán)境下氧自由基的增加所帶來的負面影響顯示心肌受損程度明顯并隨著海拔升高有升高趨勢,并且在海拔≥2260m處顯著增加；大鼠動脈血氧分壓,二氧化碳分壓,BE及HC03降低,指示在海拔≥2260m大鼠具有代償性的呼吸性堿中毒或代謝性酸中毒,并隨著海拔升高有加重趨勢。 2.對心血管系統(tǒng)的藥物進行評價發(fā)現(xiàn),在抵抗氧化損傷方面均有效,而各藥物的生化血氣結(jié)果卻各不相同,其中美托洛爾在保護心臟氧化損傷及提高動脈血氧飽合度等方面有良好的效果； 3.急進高原3400m后心肌線粒體呼吸鏈各復(fù)合體活性下降,使ROS產(chǎn)生增加,并且主要產(chǎn)生ROS部位在線粒體復(fù)合體Ⅰ、Ⅱ、Ⅲ。心肌組織氧化應(yīng)激與線粒體復(fù)合體活性的相關(guān)性分析,發(fā)現(xiàn)急進高原后氧化應(yīng)激損傷程度與線粒體復(fù)合體有顯著負相關(guān)性,因此分析線粒體復(fù)合體活性的下降是導(dǎo)致心肌組織氧化應(yīng)激損傷的主要原因,在應(yīng)用藥物美托洛爾后能通過提高線粒體復(fù)合體活性來降低ROS所帶來的急性高海拔損傷。
[Abstract]:objective
1. to 55m, 1500m, 2260m, 3400m, 4300m five altitude gradient experiment, damage situation and trend of the research on high altitude environment after myocardial in rats, to investigate the effects of oxidative stress and mitochondria in myocardial injury;
2. in oxidative stress, blood biochemical, morphological, microscopic observation and comprehensive evaluation indexes of four kinds of drug intervention effects on myocardial in rats at high altitude, the excellent effect of drug screening;
3. to explore the drug metoprolol myocardial protection at high altitude rats mechanism of oxidative damage and mitochondrial damage.
Method
Experiment one: the rats were randomly divided into 5 groups with 12 rats in each group, group A (Shanghai 55m), group B (Gansu Lanzhou 1500m), group C (Qinghai Xining 2260m), D group (group E, 3400m Gansu Luqu) (Qinghai 4300m, record the weight after Maduo) respectively from the Plains (Shanghai 55m) determination of biochemical radical groups at.3d altitude field of venous serum, measured arterial blood gas indexes, myocardial tissue fixed pathology, electron microscope observation and determination of the oxidation level for liquid nitrogen refrigeration; experiment two: radical 3400m after the four drugs were given: Amiodarone, metoprolol, Cato Plymouth, furosemide and positive the control of dexamethasone. Recorded 1D and 3D weight, 3D after the administration of rats, heart rate, blood pressure, and the orbital venous blood for biochemical tests (serum CK, CK-MB, AST, HDBH, LDH etc.) and cardiac markers (MYO, Tn-I); the experimental animal after anesthesia in abdominal aorta arterial blood gas Analysis (pH, pO2, pCO2, Hct, ctHb, cHCO3, BB, BE, SO2); record the weight of living donor rat heart tissue, cut a part fixed for pathological and electron microscopic observation; the remaining myocardial tissue using liquid nitrogen rapid freezing preservation, for the determination of the oxidation level (SOD, MDA, NO); experiment three: the plain group, the elevation of 3400m blank group and metoprolol group. Heart homogenate of mitochondrial complex (COM- I, COM- II, COM- III, COM- and IV) activity, determination of MDH. SDH and ATP activity.
Result
The first 1. groups of myocardial enzymogram of rats results in B, C, D, E group rats were significantly increased compared with group A, CK and CK-MB in B group were increased by 43.87%, 55.36%, C increased 185.14%, 84.54%, D increased 73.54%, 139.46%, E group increased 118.28% (175.32% P0.05). So the B group (1500m), C group (2260m), D group (3400m), E group (4300m) of myocardial enzymes in the blood release increased significantly; the rats cardiac biomarker results: compared with A group MYO, Tn-I in C, D, E were significantly increased. In the C group increased by 182.82%, 21.30%, D group increased by 328.35%, 54.73%, E increased 444.04%, 62.67% (P0.05), marker results show that in the C group (2260m), D group (3400m), E group (4300m) myocardial injury obviously increased compared with the plain group;
Experiment 2. compared with the A group rats showed oxidative stress in B, SOD, C, D, E were reduced by 10.06%, 13.51%, 12.18%, 29.92%; MDA in B, C, D, E group respectively increased 14.67%, 89.33%, 36.01%, 86.66%; NO in B, C, D, E groups were reduced by 64.45%, 65.49%, 66.94%, 40.54% (P0.05), C D, E in rats, myocardial oxidative stress increased, arterial oxygen pressure decreased significantly;
3. of the rat PaO2 and PaCO2 decreased significantly (P0.05) (at an altitude of more than 2260m), compared with the A group, pCO2 in B, C, D, E were 9.56%, 6.68%, 25.07%, 32.01%; pO2 in B, C, D, E were 7.95%, 11.97%, 48.94% 43.09%, S02; in D, E group were decreased respectively decreased by 12.04%, 8.25% (P0.05);
4. experiment two, on oxidative damage in different altitude effect in 3400m application of drugs, including SOD and NO compared with metoprolol group plateau blank group were increased by 10.75%, 144.65% (P0.05), MDA (P0.05), reduced by 25.49% and increase the oxygen partial pressure increased 31.68% (P0.05), can significantly reduce the spectrum increase the myocardial enzyme;
5. experiment three, at an elevation of 3400m at the mitochondrial complex I, II, III, IV activity in model group was significantly higher than that in plain plateau group decreased, the COM- I and COM- II, COM-III decreased significantly, compared with the plain group in the plateau region COM- I decreased by 26.17%, COM- decreased by 24.21% COM- II, III decreased by 17.94% COM-IV, down 32.61% (P0.05), improve the activity of drug use of metoprolol, respectively compared with the plateau group increased by 22.48%, 44.01%, 36.01%, 17.23% (P0.05), as ATP content increased significantly after the drug application (P0.05);
Experiment 6. and experiment two myocardial histopathology and electron microscopy results show that the altitude of more than 2260m compared with the plain group showed significant damage, muscle fiber is not neatly arranged, mitochondrial swelling and dissolution, intercalated disc broadening, giving drugs have different degrees of improvement, the protective effect of metoprolol was obvious.
conclusion
1. experimental animal from the plain 55m 1500m 2260m, 3400m radical, 4300m, different altitude, oxidative stress results showed that the negative influence brought by the increase of oxygen free radicals in plateau environment showed the degree of myocardial damage and significantly increased with the increase of elevation, and significantly increased at an altitude of more than 2260m; rat arterial oxygen partial pressure, the partial pressure of carbon dioxide, BE and HC03 decreased, indicating a compensatory elevation of more than 2260m in rats with respiratory alkalosis and metabolic acidosis, and with the increase of elevation has aggravated the trend.
The 2. drugs on the cardiovascular system evaluation found that are effective in resisting oxidative damage, and the blood biochemical drugs but the results are not the same, in which Mei TORO M has a good effect on the protection of cardiac oxidative damage and improve arterial oxygen saturation;
Decreased myocardial mitochondrial respiratory chain complex activity of each 3. at high altitude after 3400m, the increased production of ROS, and mainly ROS sites in the mitochondrial complex I, II, III. Correlation analysis of myocardial oxidative stress and mitochondrial complex activity, found at high altitude after oxygen had a significant negative correlation of the stress and the degree of damage of mitochondrial complex, therefore decreased analysis of mitochondrial complex activity is a major cause of oxidative stress in myocardial tissue injury, can reduce the acute high altitude ROS damage brought by increasing the activity of mitochondrial complex application in drug metoprolol.

【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R594.3

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