【摘要】：目的通過(guò)建立模擬急進(jìn)高原大鼠模型,利用HE染色、透射電鏡及免疫組化的方法,初步探討急進(jìn)高原大鼠腸損傷后細(xì)胞自噬的變化情況,為進(jìn)一步研究自噬在急進(jìn)高原相關(guān)腸道疾病中的作用提供一定理論依據(jù)。 方法將50只Wistar大鼠隨機(jī)分成5組：平原對(duì)照組,急進(jìn)高原6h、12h、24h、48h組,每組各10只。通過(guò)低壓低氧動(dòng)物實(shí)驗(yàn)艙模擬急進(jìn)海拔4767m,建立急進(jìn)高原大鼠模型。用光鏡觀察各組大鼠腸組織病理改變,用透射電鏡觀察腸上皮細(xì)胞中自噬體結(jié)構(gòu),用免疫組化法檢測(cè)自噬相關(guān)蛋白Beclinl及LC3B的表達(dá)。 結(jié)果光鏡顯示：與平原對(duì)照組比較,急進(jìn)高原6h組大鼠腸組織發(fā)生輕度損傷,急進(jìn)高原12h組及24h組損傷明顯加重,急進(jìn)高原48h組損傷最重。透射電鏡顯示：急進(jìn)高原6h組開(kāi)始發(fā)現(xiàn)雙層膜結(jié)構(gòu)的自噬體,急進(jìn)高原24h組自噬體增多最明顯,急進(jìn)高原48h組仍可見(jiàn)自噬體。免疫組化顯示：Beclinl在平原對(duì)照組,急進(jìn)高原6h、12h、24h、48h組的表達(dá)分別為(O.142±O.005)、(O.161±0.017)、(O.179±O.018)、(O.195±0.015)、(0.185±0.020):急進(jìn)高原6h、12h、24h、48h組Beclinl表達(dá)均顯著高于平原對(duì)照組(P值均0.01)。LC3B在平原對(duì)照組,急進(jìn)高原6h、12h.24h、48h組的表達(dá)分別為(0.150±O.007)、(O.171±O.015)、(0.190±0.017)、(0.207±0.016)、(O.193±0.019):急進(jìn)高原6h、12h.24h、48h組LC3B表達(dá)均顯著高于平原對(duì)照組(P值均0.01)。 結(jié)論1.急進(jìn)高原環(huán)境可使大鼠腸組織發(fā)生不同程度的損傷,并且隨著急進(jìn)高原暴露時(shí)間延長(zhǎng),腸損傷程度逐漸加重。2.急進(jìn)高原大鼠腸損傷后細(xì)胞自噬表達(dá)水平明顯增強(qiáng)。
[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 鹵0.005), (鹵0.017), (0.179 鹵0.018), (O.195 鹵0.015), (0.185 鹵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 鹵0. 007), (0. 171 鹵0. 015), (0.190 鹵0.017), (0.207 鹵0.016), (0. 193 鹵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.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R574

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