高壓電燒傷大鼠血清SOD、MDA的變化及烏司他丁的干預(yù)作用
發(fā)布時(shí)間:2018-01-16 15:14
本文關(guān)鍵詞:高壓電燒傷大鼠血清SOD、MDA的變化及烏司他丁的干預(yù)作用 出處:《河北醫(yī)科大學(xué)》2017年碩士論文 論文類型:學(xué)位論文
更多相關(guān)文章: 燒傷 高壓電 超氧化物岐化酶 丙二醛 烏司他丁 氧化應(yīng)激 大鼠
【摘要】:目的:高壓電通過電熱效應(yīng)、電化學(xué)效應(yīng)及電場(chǎng)力等作用,對(duì)機(jī)體神經(jīng)、血管、肌肉等組織及器官造成損傷。高壓電燒傷致傷的病理及生理機(jī)制尚未完全明了。研究認(rèn)為,氧化應(yīng)激參與高壓電對(duì)機(jī)體的損傷過程。研究表明,燒傷后機(jī)體產(chǎn)生大量活性氧(reactive oxygen species,ROS),活性氧通過脂質(zhì)過氧化等反應(yīng)損傷組織和器官,并與炎癥因子和血管活性物質(zhì)協(xié)同作用,造成氧化應(yīng)激損傷。高壓電燒傷導(dǎo)致全身氧化應(yīng)激反應(yīng)的因素很多,其中血漿超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)在調(diào)節(jié)氧化應(yīng)激方面有重要作用。SOD是廣泛存在于生物體的金屬氧化酶,能催化超氧陰離子自由基的歧化反應(yīng),通過清除細(xì)胞產(chǎn)生的超氧離子而發(fā)揮作用。MDA是脂質(zhì)過氧化反應(yīng)的產(chǎn)物。目前,關(guān)于高壓電燒傷后大鼠血清SOD、MDA定量及定性變化的相關(guān)研究較少,本研究檢測(cè)高壓電燒傷大鼠血清SOD、MDA含量的變化,并采用烏司他丁(Ulinastatin,UTI)進(jìn)行干預(yù),旨在探索大鼠高壓電燒傷后血清SOD、MDA的變化規(guī)律,證明氧化應(yīng)激在高壓電燒傷漸進(jìn)性損傷發(fā)生和發(fā)展過程中的作用,探索UTI對(duì)高壓電燒傷的治療效果。方法:1實(shí)驗(yàn)動(dòng)物分組:本實(shí)驗(yàn)用成年SD大鼠240只,大鼠由河北醫(yī)科大學(xué)動(dòng)物實(shí)驗(yàn)中心提供(合格證編號(hào)832457),體重291-362g,按隨機(jī)區(qū)組設(shè)計(jì)法將其隨機(jī)分為四組,每組60只,即假高壓電燒傷組(假傷組)、高壓電燒傷組(電傷組)、高壓電燒傷UTI治療組(UTI組)、高壓電燒傷鹽水治療組(鹽水組)。按隨機(jī)數(shù)字表法,每組按觀察時(shí)相分為電傷前15min、電傷后5min、電傷后1h、2h、4h、8h六個(gè)時(shí)相組,每一時(shí)相組10只大鼠。2實(shí)驗(yàn)前準(zhǔn)備:(1)將實(shí)驗(yàn)大鼠稱重并給予其編號(hào)登記;(2)準(zhǔn)備實(shí)驗(yàn)用采血管、采血針等物品,記錄實(shí)驗(yàn)數(shù)據(jù);(3)按藥品說(shuō)明配置實(shí)驗(yàn)藥品使其達(dá)到實(shí)驗(yàn)所需濃度。3高壓電燒傷動(dòng)物模型制備:用10%的水合氯醛(0.3ml/100g)溶液注入大鼠腹腔以麻醉實(shí)驗(yàn)大鼠,待麻醉成功后,將麻醉后大鼠仰臥固定于專用的電擊實(shí)驗(yàn)臺(tái)上。大鼠左上肢及右下肢脫毛處理,大鼠的左上肢(電流入口)及右下肢(電流出口)分別連接1cm×1cm電極片。將實(shí)驗(yàn)變壓器和調(diào)壓器電線相連接;再接通電源,調(diào)節(jié)調(diào)壓器指針,將升壓器輸出電壓調(diào)至2kv,接通升壓器輸出電源開關(guān),使高壓電流通過大鼠致電擊,持續(xù)電擊3s,高壓電燒傷模型制作完成。將電傷后大鼠分籠放好,待采血。假傷組不予通電,其余處理步驟與電傷組及uti組的相同。鹽水組電后5min內(nèi)注射生理鹽水5ml/kg至大鼠腹腔內(nèi),uti組電后5min內(nèi)注射uti(2ml/kg)至大鼠腹腔內(nèi)。每組樣品按時(shí)相進(jìn)行采血、分離血清。4標(biāo)本采集與保存:將高壓電燒傷大鼠再次麻醉,待麻醉后開胸暴露心臟,于心臟內(nèi)直接抽血取樣,血量約6ml,將抽出的血液注入促凝管中,靜置30min,待血清析出后,放入離心機(jī),調(diào)整轉(zhuǎn)速3000轉(zhuǎn)/min,離心10min,取上清液置于eppendorf管中于-70℃條件下保存。5指標(biāo)檢測(cè):采用酶聯(lián)免疫吸附雙抗體夾心法(enzymelinkedimmunosorbentassay,elisa),分別檢測(cè)每組大鼠的六個(gè)時(shí)相組血清sod、mda含量。6實(shí)驗(yàn)數(shù)據(jù)處理:采用spss21.0統(tǒng)計(jì)軟件,行兩因素析因設(shè)計(jì)的方差分析,多重比較采用lsd法t檢驗(yàn)。以p0.05為顯著性檢驗(yàn)水準(zhǔn)。結(jié)果:1大鼠血清sod含量變化電傷組大鼠血清sod含量與假傷組差異有顯著性,總體低于假傷組(主效應(yīng)f=165.245,p0.05);電傷組血清sod含量各時(shí)相變化差異有顯著性(主效應(yīng)f=16.905,p0.05),傷后5min~8h各時(shí)相均低于本組傷前值(p0.05),且呈逐漸降低趨勢(shì),sod含量在8h時(shí)相時(shí)達(dá)到最低,但電傷組sod含量仍較假傷組同時(shí)相低。血清sod含量uti組與鹽水組差異有顯著性,總體高于鹽水組(主效應(yīng)f=31.043,p0.05);uti組sod含量各時(shí)相變化差異有顯著性(主效應(yīng)f=62.830,p0.05),uti組sod含量傷后5min~8h各時(shí)相均高于本組傷前值(p0.05),且呈逐漸降低趨勢(shì)。2大鼠血清mda含量變化電傷組血清mda含量與假傷組差異有顯著性,總體高于假傷組(主效應(yīng)F=1586.203,P0.05);電傷組MDA含量各時(shí)相變化差異有顯著性(主效應(yīng)F=126.552,P0.05),傷后5min~8h呈逐漸增高趨勢(shì),且各時(shí)相均高于本組電傷前值(P0.05)。UTI組血清MDA含量與鹽水組差異有顯著性,總體低于鹽水組(主效應(yīng)F=45.326,P0.05);UTI組MDA含量各時(shí)相變化差異有顯著性(主效應(yīng)F=246.201,P0.05),UTI組傷后5min~8h各時(shí)相均高于本組傷前值(P0.05),且呈逐漸增高趨勢(shì)。結(jié)論:高壓電燒傷使大鼠血清SOD含量降低、MDA含量升高,誘導(dǎo)高壓電燒傷后氧化應(yīng)激反應(yīng)。高壓電燒傷后采用UTI治療,可使血清SOD的含量增加,MDA含量降低,表明UTI可以抑制高壓電燒傷后的氧化應(yīng)激反應(yīng),減輕氧化應(yīng)激對(duì)高壓電燒傷后機(jī)體的損傷。
[Abstract]:Objective: through high-voltage electric heating effect, effect of electrochemical effect and electric field force on the body, vascular nerve, muscle damage to tissues and organs. High voltage electric burn injury pathological and physiological mechanisms are not completely understood. Studies suggest that oxidative stress is involved in the damage process of high-voltage electrical machine body. The results show that after burn the body produces a large number of reactive oxygen species (reactive oxygen, species, ROS), reactive oxygen species through lipid oxidation reaction of damaged tissue and organs, and the synergistic effect of vasoactive substances and factors, inflammation, oxidative stress injury caused by high voltage electric burn. Factors leading to systemic oxidative stress reaction of many, including plasma superoxide dismutase (superoxide dismutase SOD, malondialdehyde (malondialdehyde), and MDA) in the regulation of oxidative stress plays an important role in metal.SOD oxidase widely exists in organisms, can catalyze the superoxide anions By the disproportionation reaction of radicals, superoxide anion scavenging by cells and play a role of.MDA is the product of lipid peroxidation. At present, on serum SOD of rats after electric burns, MDA related research on quantitative and qualitative changes less, this study is to detect the high-voltage electric burn rat serum SOD, MDA content changes, and the effects of ulinastatin (Ulinastatin, UTI) to intervene, to explore after electrical burn serum SOD hypertension rats, the changing rules of MDA, prove the oxidative stress in the high voltage electric burn progressive damage occurrence and development process, explore the therapeutic effect of UTI on high voltage electric burn. Methods: 1 animal groups: experiment with 240 adult SD rats, rats were provided by the animal experimental center of Hebei Medical University (Certificate No. 832457), weight 291-362g, according to a randomized block design method will be divided into four groups randomly, 60 rats in each group: sham, high-voltage electric burn 緇,
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