本文選題：低氧 + 端粒�。� 參考：《青海大學》2017年碩士論文

【摘要】：目的:通過檢測急性低氧環(huán)境下大鼠胃組織細胞端粒長度及TERT(端粒酶逆轉錄酶,Telomerase reverse transcriptase)、HIF-1a(低氧誘導因子1α,Hypoxia-inducible factor 1α)、HIF-2a(低氧誘導因子2α,Hypoxia-inducible factor2α)、ROS(活性氧自由基,reactive oxygen species)表達水平,觀察急性低氧環(huán)境下大鼠胃組織細胞端粒長度的變化,并探討其具體機制。方法:(1)選擇雄性Wistar大鼠40只,隨機分為對照組(n=10,蘭州,1500米)與實驗組(n=30,低壓氧艙,5000米),實驗組根據(jù)低氧暴露時間不同,分為低氧1d、3d、7d三組,每組各10只,取各組大鼠胃組織,行HE染色并在顯微鏡下觀察胃組織細胞大體形態(tài)結構變化;(2)提取各組大鼠胃組織細胞DNA,采用Real-time PCR法測其端粒長度;(3)提取各組大鼠胃組織細胞總RNA及蛋白,采用Real-time PCR及Elisa法分別測其TERT、HIF-1α及HIF-2αm RNA及蛋白表達水平;(4)制備各組大鼠胃組織勻漿液,采用化學熒光法測其ROS水平;(5)分析各組大鼠胃組織細胞TERT、HIF-1α、HIF-2α及ROS水平在其端粒長度變化中的作用,并行相關性分析。結果:(1)在低壓氧艙中模擬5000m高海拔(嚴重低氧環(huán)境)構建出高原嚴重低氧對大鼠胃組織細胞的損傷模型,其病理結果顯示隨著低氧時間的逐漸延長,胃黏膜損傷逐漸加重,主要表現(xiàn)為胃黏膜腺體逐漸萎縮,黏膜充血,大量炎細胞浸潤,細胞發(fā)生嗜酸性變;(2)與對照組相比,各時間點實驗組端粒長度均延長(P0.05),并且隨著低氧時間的延長,端粒長度表現(xiàn)為逐漸增加的趨勢;(3)各組TERT、HIF-1αm RNA及蛋白表達水平的變化趨勢與其端粒長度的變化趨勢基本一致,但其HIF-2αm RNA及蛋白表達水平未見這一變化趨勢,僅在低氧7天組表達增加;(4)實驗組ROS表達量與對照組相比均增加(P0.05),并且隨著低氧時間的延長,ROS表達量逐漸升高;(5)大鼠胃組織細胞中TERT與HIF-1α的蛋白表達水平呈正相關(r=0.839 p0.05),而TERT與HIF-2α蛋白表達水平無相關性(r=0.298 p=0.062);(6)大鼠胃組織細胞中TERT與HIF-1αm RNA表達水平呈正相關(r=0.904 p0.05);TERT m RNA表達水平與端粒長度呈正相關(r=0.962 p0.05),而TERT與HIF-2αm RNA表達無相關性(r=0.298 p=0.248)。結論:(1)急性嚴重高原低氧環(huán)境可使機體ROS表達增加,進而導致大鼠胃組織細胞損傷;(2)急性嚴重高原低氧環(huán)境可誘導大鼠胃組織細胞端粒延長,并且端粒的長度隨著低氧時間的延長而延長,且與TERT與HIF-1α表達呈正相關,而與HIF-2α表達無明顯相關,由此考慮急性高原嚴重低氧引起端粒延長的機制為:大鼠胃組織細胞HIF-1α表達水平升高,誘導TERT表達及端粒酶活性增加,來延長端粒;(3)急性嚴重高原低氧環(huán)境下,大鼠胃組織細胞可以通過延長端粒,來對抗低氧對其的損害,不致于使其發(fā)生嚴重的致命性損傷。
[Abstract]:Objective: to detect the length of telomere and the expression of reactive oxygen species) (reactive oxygen species) of reactive oxygen free radical) in gastric tissue cells of rats exposed to acute hypoxia, and to detect the expression of HIF-2a (Hypoxia-inducible factor 1 偽) and telomerase reverse transcriptase (reverse transcriptase) HIF-1a (hypoxia inducible factor 1 偽). To observe the changes of telomere length in gastric tissue of rats under acute hypoxia and to explore its mechanism. Methods: (1) Forty male Wistar rats were randomly divided into control group (n = 10, Lanzhou 1500m) and experimental group (n = 30, hypobaric oxygen chamber, 5000m). According to the time of hypoxia exposure, the experimental group was divided into three groups with 10 rats in each group. He staining was performed and observed under microscope. (2) DNA of gastric tissue cells was extracted from each group and telomere length was measured by Real-time PCR. (3) Total RNA and protein of gastric tissue cells were extracted from each group. Real-time PCR and Elisa were used to detect the mRNA and protein expression levels of TERTHIF-1 偽 and HIF-2 偽, respectively. (4) the gastric tissue homogenate of rats in each group was prepared. (5) the role of TERTHIF-1 偽 HIF-2 偽 and Ros in telomere length was analyzed. Results: (1) the model of gastric tissue cells injury induced by severe hypoxia at altitude of 5000m was established in the hypobaric oxygen chamber. The pathological results showed that the gastric mucosal injury was aggravated with the prolongation of hypoxia time. The main manifestations of gastric mucosal gland atrophy, mucosal congestion, a large number of inflammatory cells infiltration, eosinophilic cells; (2) compared with the control group, the telomere length of the experimental group was prolonged at each time point (P0.05), and with the prolongation of hypoxic time, Telomere length showed a tendency of increasing gradually. (3) the change trend of telomere length was basically the same as that of telomere length, but the expression level of HIF-2 偽 mRNA and protein was not. (4) the expression of Ros in the experimental group was higher than that in the control group (P0.05), and gradually increased with the prolongation of hypoxia time. (5) the expression level of TERT and HIF-1 偽 protein in gastric tissue cells of rats was positive. The expression level of TERT and HIF-1 偽 mRNA was positively correlated with telomere length (r 0.962 p0.05) in rat gastric tissue cells (r 0.298pn 0.062); (6), while TERT was not correlated with HIF-2 偽 mRNA expression (r 0.298pt0. 248) in rat gastric tissue cells (r = 0.904 p0.05), while TERT was not correlated with HIF-2 偽 mRNA expression (r = 0.298pt0. 248). The expression of TERT mRNA was positively correlated with telomere length (rr 0.962 p0.05), but not between TERT and HIF-2 偽 mRNA expression (r = 0.298p0. 248). Conclusion: (1) acute severe high altitude hypoxia can increase the expression of Ros and induce gastric tissue cell injury in rats, (2) acute severe high altitude hypoxia can induce telomere prolongation in gastric tissue of rats. The length of telomere increased with the prolongation of hypoxia time, and was positively correlated with the expression of TERT and HIF-1 偽, but not with the expression of HIF-2 偽. Therefore, the mechanism of telomere prolongation caused by severe hypoxia at high altitude is as follows: the expression of HIF-1 偽 in rat gastric tissue cells is increased, and the expression of TERT and telomerase activity are increased to prolong telomere; (3) in acute severe hypoxia environment, the expression of TERT and telomerase activity are increased in order to prolong telomere. Rat gastric tissue cells can prolong telomeres to prevent hypoxia from causing severe and fatal damage.
【學位授予單位】：青海大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R594.3

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