【摘要】 目的 觀察長期大量酒精攝入對大鼠心肌結(jié)構(gòu)及心肌組織中丙二醛(MDA)、超氧化物歧化酶(SOD)和金屬硫蛋白(MT)含量的影響,探討氧化應(yīng)激在酒精性心肌病大鼠中的作用。 方法 雄性健康SD大鼠45只,隨機分為2組,即對照組20只和模型組25只。模型組酒精濃度從5%、10%、20%和30%依次各自由飲1周,然后遞增至36%后以該濃度維持飼喂。對照組每日飲用與模型組酒精同等熱量的葡萄糖水。6個月后,觀察大鼠心肌組織的形態(tài)學(xué)改變及超微結(jié)構(gòu)的變化,測定心肌組織中MDA、SOD及MT的含量。結(jié)果 模型組大鼠心肌細胞排列紊亂、間質(zhì)充血、炎細胞浸潤、線粒體腫脹、空泡形成、肌絲溶解、核膜不規(guī)則和核仁裂解。心肌組織中MDA含量明顯升高(Plt;0.01),SOD活力含量明顯降低(Plt;0.01),MT含量明顯降低(Plt;0.01)。 結(jié)論 長期攝入大量酒精可使氧自由基代謝失衡,導(dǎo)致心肌損傷。氧化應(yīng)激在酒精性心肌病發(fā)病機制中發(fā)揮著重要的作用。【Abstract】 Objective To observe the effect of longterm and large quantities of alcohol intake on myocardial structure of rats and the content of malondialdehyde (MDA), superoxide dismutase (SOD) and metallothionein (MT) in myocardium tissue. To study the effect of oxidative stress on the rats with alcoholic cardiomyopathy. Methods Fortyfive male and healthy SD rats were randomly divided into the control group (20 rats) and model group (25 rats).The alcoholic concentrate in model group was increased from 5%,10%,20% to 30% every week, and maintain free drinking mass concentration of 36% alcohol. The control group drink the same calories of glucose water. Six months later, the myocardial tissues were observed both in light microscope and electron microscope .The level of MDA、SOD and MT were tested in myocardium tissue. Results In the model rats, the cells of myocardial disarray, interstitial congestion, inflammatory cell infiltration, mitochondrial swelling, vacuole formation, melt filaments, irregular nuclear membrane and nucleolus cracking. The content of MDA incresed(Plt;0.01)and the activities of SOD decreased(Plt;001),levels of MT decreased (Plt;0.01) in the cardiac muscular tissues in the model group compared with the control group. Conclusion Longterm intake of large amounts of alcohol can break the balance of oxygen free radicals, which leading to the damage of myocardial. Oxidative stress plays an important role in the etiopathogenesis of alcoholic cardiomyopathy.
目的觀察高糖誘導(dǎo)的牛視網(wǎng)膜血管內(nèi)皮細胞(BRECs)中共濟失調(diào)毛細血管擴張突變基因(ATM)激酶的活性變化及其對細胞氧化應(yīng)激狀態(tài)的影響。 方法體外培養(yǎng)BRECs, 取4~8代細胞用于實驗。采用含5 mmol/L葡萄糖(正常糖組)、30 mmol/L葡萄糖(高糖組)、30 mmol/L葡萄糖和10 μmol/L ATM激酶特異性抑制劑KU55933(干預(yù)組)的細胞培養(yǎng)液培養(yǎng)BRECs。細胞培養(yǎng)48 h后, 采用蛋白免疫印跡法檢測細胞中磷酸化ATM(P-ATM)、磷酸化細胞分裂素活化蛋白激酶(P-P38)及磷酸化細胞外信號調(diào)節(jié)激酶(P-ERK)1/2的蛋白表達水平; 酶聯(lián)免疫吸附試驗檢測細胞上清液中血管內(nèi)皮生長因子(VEGF)的含量; 細胞內(nèi)活性氧(ROS)水平檢測試劑盒檢測細胞內(nèi)ROS水平; 碘化丙啶和赫斯特染料雙染色法檢測細胞凋亡情況; 異硫氰酸熒光素標(biāo)記的葡聚糖檢測內(nèi)皮細胞屏障的通透性。 結(jié)果與正常糖組比較, 高糖組P-ATM、P-P38、P-ERK1/2蛋白表達均增高; 與高糖組比較, 干預(yù)組P-ATM蛋白表達降低, p-p38、p-ERK1/2蛋白表達明顯增加。3組間P-ATM、P-P38、P-ERK1/2蛋白表達比較, 差異均有統(tǒng)計學(xué)意義(F=436.00、14 010.00、985.10, P＜0.05)。與正常糖組比較, 高糖組、干預(yù)組細胞上清液中VEGF含量均升高, 干預(yù)組升高幅度更明顯; 3組間細胞上清液中VEGF含量比較, 差異有統(tǒng)計學(xué)意義(F=278.00, P＜0.05)。高糖組ROS含量(t=2.98、10.91)、BRECs細胞凋亡率(t=4.31、11.14)均較正常糖組升高, 而又明顯低于干預(yù)組, 差異均有統(tǒng)計學(xué)意義(P＜0.05)。與正常糖組比較, 高糖組、干預(yù)組細胞旁通透率均升高, 干預(yù)組升高更明顯; 3組間細胞旁通透率比較, 差異有統(tǒng)計學(xué)意義(F=2 223.00, P＜0.05)。 結(jié)論高糖誘導(dǎo)的BRECs中P-ATM表達、ROS水平、細胞凋亡率及細胞旁通透率均增高, 其機制可能與P38、ERK、VEGF有關(guān)。