新生大鼠高氧动物模型肠道AKT的变化  

Intestinal AKT Expression in Neonatal Rats with Hyperoxia

作　　者：王思[1] 曹旭[1] 刘冬妍[1] 

机构地区：[1]中国医科大学附属盛京医院实验研究中心,卫生部小儿先天畸型重点实验室,沈阳110004

出　　处：《中国医科大学学报》2013年第7期577-581,共5页Journal of China Medical University

基　　金：国家自然科学基金(30871158,81170604);盛京自由研究者基金

摘　　要：目的研究高氧对新生大鼠肠道AKT信号通路的影响。方法将新生大鼠分为高氧组和空气组,采用电镜观察2组大鼠肠组织结构的变化;应用免疫组化染色(SP法)和Western blot检测肠组织AKT和p-AKT蛋白的表达;应用实时PCR检测肠组织AKT mRNA的表达。结果电镜显示空气组肠黏膜形态完整;高氧组可见上皮表面绒毛有部分缺失,核形不规则,胞质内疑有凋亡小体。免疫组化结果显示,空气组AKT蛋白表达随生长发育逐渐增加;与空气组比较,高氧组AKT表达第7天升高,第10天反而下降(P<0.05)。空气组p-AKT蛋白表达亦随生长发育逐渐增加,与空气组比较高氧组p-AKT表达第7天明显升高(P<0.05),但第10天表达减少。Western blot结果显示,与空气组比较,高氧组AKT表达第7天显著升高(P<0.05),但第10天下降。p-AKT表达亦升高,但差异无统计学意义(P>0.05)。实时PCR结果显示,与空气组比较,高氧组AKT mRNA表达第5天和第7天显著升高(P<0.05),但在第10天表达下降(P<0.05)。结论高氧刺激使肠道黏膜细胞结构发生改变,肠道黏膜细胞可能通过AKT的磷酸化来对抗高氧刺激进而保护肠道黏膜。Abstract Objective To explore the effect of hyperoxia on intestinal AKT of neonatal rats. Methods Electron microscopy was used to observe the structure change of rat intestinal tissues. Immunohistochemistry staining and Western blot were applied to detect the protein expression of AKT and p-AKT. Real-time PCR was employed to evaluate the mRNA level of AKT. Results Electron microscopy results showed an integrity structure of rat intestinal cells in the air group; while the hyperoxia group showed that parts of villus were absence, irregular nuclear and apoptosis body was increased. Compared with those of the air group, the expression of AKT and p-AKT was significantly increased in the 7th day, but decreased in the 10th day （P 〈 0.05 ）. The change of p-AKT was similar with AKT （P 〈 0.05 ）. The results of Western blot showed that the expression of AKT in hyperoxia group was higher than the air group except the 10th day and was significantly increased in the 7th day （P 〈 0.05 ）. The expression of p-AKT increased but there was no significantly difference. The result of real-time PCR showed that the expression level of AKT in hypemxia group was higher than that of the air group, but significant differences were only observed in the 7th and 10th day. Conclusion In hyperoxia, the structure of intestinal mucosa was changed, which affected the function of intestinal mucosa. The intestinal mucosal ceils might against hypemxia stimulation through AKT phosphorylation, which further protects the intestinal mucosa.

关 键 词：高氧 肠道 蛋白激酶B 磷酸化的蛋白激酶B 

分 类 号：R392[医药卫生—免疫学]