急性低氧环境下HSP72蛋白在大鼠肾脏中的表达及意义

发布时间：2018-01-22 05:48

本文关键词： 急性低氧肾脏热休克蛋白72　出处：《青海大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：利用低压氧舱模拟急性低氧环境，复制急性缺氧动物模型，探讨急性低氧环境下大鼠肾脏热休克蛋白72（heat shock protein,HSP72）的表达及意义。材料与方法：30只健康雄性SD大鼠，体重均在180g~220g之间，随机分为急性低氧组，分别为：12h、24h、48h、72h、1周组，模拟海拔高度为5000m，同时设立对照组（西宁地区）。SD大鼠适应性喂养2周后，将其中五组大鼠放入低压氧舱建立急性低氧动物模型，，一组放于动物房普通饮食饲养。分别在不同时间段从低压氧舱内取出各组大鼠，即刻进行取材：20%乌拉坦（1ml/100g）经腹腔注入麻醉大鼠后，充分暴露腹腔，摘取双侧肾脏。大鼠肾脏组织用蛋白免疫印迹（Western Blotting）法检测HSP72蛋白的表达水平；同时膀胱穿刺留取各组大鼠尿液，离心后吸取上清液，用酶联免疫吸附测定法（ELISA法）检测大鼠尿微量白蛋白及尿肌酐浓度。结果：低压氧舱内模拟大鼠急性缺氧后，可见各低氧组大鼠肾组织中HSP72蛋白的表达水平明显高于对照组，而对照组几乎很少表达，随着缺氧时间的延长，大鼠肾组织中HSP72蛋白的表达水平逐渐增高，于72h达最高水平（2.68±0.08），到1周时其表达水平有所下降（1.59±0.17），各组与对照组（0.19±0.12）比较差异均有统计学意义（P0.01）；分别测定各组大鼠尿微量白蛋白及尿肌酐的含量，然后求其比值，结果提示尿微量白蛋白与尿肌酐比值从48h开始升高，48h（235.40±20.60）、72h（378.12±61.31）、1周组（504.55±48.77）与对照组（169.63±17.45）比较差异有统计学意义（P0.05）。结论：急性低氧可以引起大鼠尿微量白蛋白排泄量增加，并随着缺氧时间的延长而增加，同时伴有肾脏HSP72蛋白表达的增加，但二者表达呈现不一致性。提示缺氧是引起肾脏损伤的重要原因，虽然HSP72升高是机体保护性应激反应，但在急性缺氧时，并不能充分缓解缺氧引起的脏器损伤，其保护作用可能具有一定的滞后性。
[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 卤0.08 at 72 h. At 1 week, the expression level decreased by 1.59 卤0.17, and there was significant difference between each group and the control group (0.19 卤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 卤20.60 ~ 72h ~ 378.12 卤61.31). There was significant difference between the 1 week group (504.55 卤48.77) and the control group (169.63 卤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.
【学位授予单位】：青海大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R594.3

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