| Objective:To observe Animal model of acute hypoxia Wistar ratsplateau autophagy inhibitor3-MA after the intervention,the organization form of intestinal mucosal epithelial injury after the change,the expression of Tight binding protein occludin and occludin m RNA,the expression of autophagy function state Beclin1,Beclin1 m RNA,LC3,LC3 m RNA,to explore whether autophagy has protective effect on intestinal barrier function under acute high altitude hypoxia.Methods: Twenty-four healthy male Wistar rats with a body weight of about(200±20)g and aged 7-8 weeks were randomly divided into the control group,plateau hypoxia group and plateau hypoxia 3-MA group.The acute plateau hypoxia rat model was established under the simulated environment of low-pressure oxygen in the plateau low-pressure oxygen chamber.The optimal concentration of 3-methyladenine(3mg/kg)and the optimal altitude(6000m,48h)were selected in the previous and preliminary experiment,and the selected concentration of3-ma(3mg/kg)were intraperitoneally injected in the plateau hypoxia 3-MA group half an hour before the oxygen chamber,the control group and the plateau hypoxia group were intraperitoneally injected with Physiological saline.The behavioral changes of rats in each group were observed.HE staining was used to analyze the damage degree of rat intestinal tissue.The expressions of Occludin,autophagy related proteins(LC3,Beclin-1)and LC3,Beclin-1 and Occludin m RNA were detected by Western blot and RT-PCR.Results:(1)HE staining of intestinal mucosa in rats with acute hypoxia at altitude of 6000 m and 48 h showed that the intestinal villi in the plateau group showed edema,shedding,disorder in arrangement,thinning of intestinal wall,and even separation of epidermis and lamina propria on both sides of intestinal villi.Compared with the plateau hypoxia group,intestinal tissue of the 3-MA group was damage more seriouse,It can be seen that with local separation of mucosa and submucosa,and even lamina propria nudity and inflammatory granulocyte aggregation.(2)Rt-PCR analysis showed that,compared with the control group,the expressions of autophagy related genes LC3 and Beclin-1 m RNA in the plateau hypoxia group were increased(P<0.01),with statistical difference.Compared with the plateau hypoxia group,LC3 and Beclin-1 m RNA expressions in intestinal mucosa were decreased in the 3-MA group,and the differences between the two groups were statistically significant(P<0.01).In addition,Occludin m RNA expression in intestinal mucosa decreased gradually in the control group,plateau hypoxia group and plateau 3-MA group(control group> plateau hypoxia group>3-methyladenine group),and with statistically significant differences(P<0.01).(3)Western Blot analysis showed that compared with the control group,LC3 and Beclin-1 protein expressions in intestinal mucosa were increased in the plateau group(P<0.01),with statistical difference.Compared with the plateau hypoxia group,LC3 and Beclin-1 protein expressions in the 3-MA group decreased significantly(P<0.01).The expression level of Occludin protein in the three groups was consistent with the expression trend of Occludin m RNA,and there were significant statistical differences among the groups(P<0.05).Conclusion :Through the establishment of simulation animal model of acute plateau hypoxia found acute plateau hypoxia not only have a harm effect on intestinal mucosa barrier,but also autophagy inhibitor 3-MA can be tied to cut the expression of intestinal mucosa epithelial cells associated protein Occludin and autophagy related genes LC3,Beclin 1,It is suggested that autophagy may play a protective role in the process of intestinal barrier function injury in rats with acute hypoxia at high altitude. |
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