急进高原大鼠肠损伤后自噬变化的初步研究

发布时间：2018-10-25 12:53

【摘要】：目的通过建立模拟急进高原大鼠模型,利用HE染色、透射电镜及免疫组化的方法,初步探讨急进高原大鼠肠损伤后细胞自噬的变化情况,为进一步研究自噬在急进高原相关肠道疾病中的作用提供一定理论依据。方法将50只Wistar大鼠随机分成5组：平原对照组,急进高原6h、12h、24h、48h组,每组各10只。通过低压低氧动物实验舱模拟急进海拔4767m,建立急进高原大鼠模型。用光镜观察各组大鼠肠组织病理改变,用透射电镜观察肠上皮细胞中自噬体结构,用免疫组化法检测自噬相关蛋白Beclinl及LC3B的表达。结果光镜显示：与平原对照组比较,急进高原6h组大鼠肠组织发生轻度损伤,急进高原12h组及24h组损伤明显加重,急进高原48h组损伤最重。透射电镜显示：急进高原6h组开始发现双层膜结构的自噬体,急进高原24h组自噬体增多最明显,急进高原48h组仍可见自噬体。免疫组化显示：Beclinl在平原对照组,急进高原6h、12h、24h、48h组的表达分别为(O.142±O.005)、(O.161±0.017)、(O.179±O.018)、(O.195±0.015)、(0.185±0.020):急进高原6h、12h、24h、48h组Beclinl表达均显著高于平原对照组(P值均0.01)。LC3B在平原对照组,急进高原6h、12h.24h、48h组的表达分别为(0.150±O.007)、(O.171±O.015)、(0.190±0.017)、(0.207±0.016)、(O.193±0.019):急进高原6h、12h.24h、48h组LC3B表达均显著高于平原对照组(P值均0.01)。结论1.急进高原环境可使大鼠肠组织发生不同程度的损伤,并且随着急进高原暴露时间延长,肠损伤程度逐渐加重。2.急进高原大鼠肠损伤后细胞自噬表达水平明显增强。
[Abstract]:Objective to study the changes of autophagy after intestinal injury in rats at high altitude by using HE staining, transmission electron microscopy and immunohistochemistry. It provides a theoretical basis for further study on the role of autophagy in acute plateau-related intestinal diseases. Methods Fifty Wistar rats were randomly divided into 5 groups: plain control group (n = 10) and control group (n = 10). A rat model of rapid entry into plateau was established by simulating the rapid advance altitude of 4767 m in the experimental chamber of low pressure and hypoxia. The pathological changes of intestinal tissue were observed by light microscope, the structure of autophagy in intestinal epithelial cells was observed by transmission electron microscope, and the expression of autophagy associated protein Beclinl and LC3B were detected by immunohistochemical method. Results compared with the plain control group, the intestinal tissue of the rats in the 6h group was slightly damaged, the injury was aggravated in the 12h group and 24h group, and the damage was the most serious in the 48h group. Transmission electron microscopy (TEM) showed that autophagy with double-layer membrane structure was found at 6h of rapid entry to high altitude, and the increase of autophagy was most obvious in 24h group, and autophagy was still observed in 48h group. Immunohistochemical staining showed that the expression of Beclinl in the plain control group and in the acute altitude group at 6h, 12h and 24h were significantly higher than those in the plain control group (0.142 卤0.005), (卤0.017), (0.179 卤0.018), (O.195 卤0.015), (0.185 卤0.020), and the Beclinl expression in the acute high altitude group (6h / 12h / 24h) was significantly higher than that in the plain control group (P < 0.01), and the expression of Beclinl in the plain control group was significantly higher than that in the plain control group (P < 0.05). The expression of LC3B was significantly higher than that of the plain control group (0.150 卤0. 007), (0. 171 卤0. 015), (0.190 卤0.017), (0.207 卤0.016), (0. 193 卤0.019), and the expression of LC3B was significantly higher than that of the plain control group (P < 0. 01). Conclusion 1. Acute exposure to high altitude could cause different degree of intestinal injury in rats, and the degree of intestinal injury increased gradually with the prolongation of exposure time. 2. The expression of autophagy was significantly increased after intestinal injury in rats at high altitude.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R574

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