發(fā)布時(shí)間：2018-01-22 05:48

  本文關(guān)鍵詞： 急性 低氧 腎臟 熱休克蛋白72　出處：《青海大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：利用低壓氧艙模擬急性低氧環(huán)境，復(fù)制急性缺氧動(dòng)物模型，探討急性低氧環(huán)境下大鼠腎臟熱休克蛋白72（heat shock protein,HSP72）的表達(dá)及意義。 材料與方法：30只健康雄性SD大鼠，體重均在180g~220g之間，隨機(jī)分為急性低氧組，分別為：12h、24h、48h、72h、1周組，模擬海拔高度為5000m，同時(shí)設(shè)立對(duì)照組（西寧地區(qū)）。SD大鼠適應(yīng)性喂養(yǎng)2周后，將其中五組大鼠放入低壓氧艙建立急性低氧動(dòng)物模型，，一組放于動(dòng)物房普通飲食飼養(yǎng)。分別在不同時(shí)間段從低壓氧艙內(nèi)取出各組大鼠，即刻進(jìn)行取材：20%烏拉坦（1ml/100g）經(jīng)腹腔注入麻醉大鼠后，充分暴露腹腔，摘取雙側(cè)腎臟。大鼠腎臟組織用蛋白免疫印跡（Western Blotting）法檢測(cè)HSP72蛋白的表達(dá)水平；同時(shí)膀胱穿刺留取各組大鼠尿液，離心后吸取上清液，用酶聯(lián)免疫吸附測(cè)定法（ELISA法）檢測(cè)大鼠尿微量白蛋白及尿肌酐濃度。 結(jié)果：低壓氧艙內(nèi)模擬大鼠急性缺氧后，可見(jiàn)各低氧組大鼠腎組織中HSP72蛋白的表達(dá)水平明顯高于對(duì)照組，而對(duì)照組幾乎很少表達(dá)，隨著缺氧時(shí)間的延長(zhǎng)，大鼠腎組織中HSP72蛋白的表達(dá)水平逐漸增高，于72h達(dá)最高水平（2.68±0.08），到1周時(shí)其表達(dá)水平有所下降（1.59±0.17），各組與對(duì)照組（0.19±0.12）比較差異均有統(tǒng)計(jì)學(xué)意義（P0.01）；分別測(cè)定各組大鼠尿微量白蛋白及尿肌酐的含量，然后求其比值，結(jié)果提示尿微量白蛋白與尿肌酐比值從48h開(kāi)始升高，48h（235.40±20.60）、72h（378.12±61.31）、1周組（504.55±48.77）與對(duì)照組（169.63±17.45）比較差異有統(tǒng)計(jì)學(xué)意義（P0.05）。 結(jié)論：急性低氧可以引起大鼠尿微量白蛋白排泄量增加，并隨著缺氧時(shí)間的延長(zhǎng)而增加，同時(shí)伴有腎臟HSP72蛋白表達(dá)的增加，但二者表達(dá)呈現(xiàn)不一致性。提示缺氧是引起腎臟損傷的重要原因，雖然HSP72升高是機(jī)體保護(hù)性應(yīng)激反應(yīng)，但在急性缺氧時(shí)，并不能充分緩解缺氧引起的臟器損傷，其保護(hù)作用可能具有一定的滯后性。
[Abstract]:Objective: to establish an acute hypoxic animal model by using hypobaric oxygen chamber to simulate acute hypoxic environment, and to study the heat shock protein 72 shock protein in the kidney of rats under acute hypoxia. HSP72) and its significance. Materials and methods 30 healthy male Sprague-Dawley rats, weighing between 180 g and 220 g, were randomly divided into acute hypoxia group (n = 12) and control group (n = 12). The simulated altitude was 5 000 m.The control group (Xining area SD rats were fed adaptively for 2 weeks) was put into hypobaric oxygen chamber to establish acute hypoxic animal model. One group was fed in a common diet in the animal room. Rats in each group were taken out from the hypobaric oxygen chamber at different time periods, and then the rats were injected intraperitoneally into anesthetized rats with 1 ml / 100g of 1 ml / 100g of 1 ml / 100 g of 1% uratan. The expression of HSP72 protein was detected by Western blotting method. At the same time, urine samples were collected from rats by bladder puncture, supernatants were collected after centrifugation, and urine microalbumin and creatinine concentrations in rats were detected by enzyme-linked immunosorbent assay (Elisa). Results: after acute hypoxia in hypobaric oxygen chamber, the expression of HSP72 protein in renal tissue of rats in hypoxia group was significantly higher than that in control group, but the expression of HSP72 protein in control group was almost rare. With the prolongation of hypoxia time, the expression of HSP72 protein increased gradually and reached the highest level of 2.68 鹵0.08 at 72 h. At 1 week, the expression level decreased by 1.59 鹵0.17, and there was significant difference between each group and the control group (0.19 鹵0.12). The contents of urinary microalbumin and urinary creatinine were measured and the ratio of urinary microalbumin to urinary creatinine was calculated. The results showed that the ratio of urinary microalbumin to urinary creatinine increased from 48h. 48h ~ 235.40 鹵20.60 ~ 72h ~ 378.12 鹵61.31). There was significant difference between the 1 week group (504.55 鹵48.77) and the control group (169.63 鹵17.45). Conclusion: acute hypoxia can induce the increase of urinary microalbumin excretion in rats, and increase with the prolongation of hypoxia time, accompanied by the increase of HSP72 protein expression in kidney. However, the expression of these two proteins is inconsistent. It is suggested that hypoxia is an important cause of renal injury. Although the increase of HSP72 is a protective stress response, it is during acute hypoxia. It can not fully alleviate the organ injury caused by hypoxia, and its protective effect may have some lag.
【學(xué)位授予單位】：青海大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R594.3

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