本文選題：溺死 + ACTB��； 參考：《重慶醫(yī)科大學》2016年碩士論文

【摘要】：目的探討溺死大鼠死后不同時間點mRNA的變化及穩(wěn)定性,篩選穩(wěn)定的內(nèi)參基因,并定量水通道蛋白5(Aquaporin,AQP-5)mRNA的表達。方法選取30只無特定病原體(Specific pathogen Free,SPF)級健康成年SD(Sprague Dawley)大鼠,并按照隨機單位組設計分組法,以每組6只,隨機均勻分為對照組(脊椎脫臼法處死)、溺死15min組(溺死尸體置于水中15min),溺死4h組(溺死尸體置于水中4h),溺死16h組(溺死尸體置于水中16h),溺死48h組(溺死尸體置于水中48h)。取溺死尸體及對照組尸體解剖,觀察肺臟變化,并分取肺組織,一部分用4%多聚甲醛(polyformaldehyde,PFA)中性PBS(phosphate buffer saline)液固定,用于形態(tài)學檢測;一部分液氮速凍,提取RNA,采用qPCR(Real Time PCR,實時熒光定量聚合酶鏈反應)篩選穩(wěn)定的內(nèi)參基因以及對AQP-5進行定量檢測。結果與對照組比較,各溺死組大鼠肺部均呈嚴重水腫,一定程度破裂出血,并可感覺到明顯溺液存于肺臟。通過提取RNA,并行q PCR,geNorm和NormFinder排序篩選發(fā)現(xiàn)ACTB(β-Actin,β-肌動蛋白)為最穩(wěn)定的內(nèi)參基因;檢測AQP-5的表達我們發(fā)現(xiàn)溺死大鼠較對照組大鼠AQP-5相對表達量下降。結論篩選出ACTB為最穩(wěn)定的內(nèi)參基因,各溺死時間點大鼠AQP-5 mRNA的表達均顯著下調(diào)。本研究中AQP-5 mRNA檢測在大鼠溺死表達有一定陽性意義,有助于法醫(yī)溺死鑒定。
[Abstract]:Objective to investigate the changes and stability of mRNA at different time points after drowning in rats, to screen stable internal reference genes and to quantify the expression of aquaporin aquaporin AQP-5 mRNA.Methods Thirty healthy adult SD(Sprague Dawley rats without specific pathogen were selected, and 6 rats in each group were divided into two groups according to random unit group.They were randomly divided into control group (spinal dislocated method, 15min group, drowning dead body in water for 15 minutes, drowning dead body in water for 4 hours, drowning dead body in water for 16 hours, drowning dead body in water for 16 hours and drowning dead body in water for 48 hours.The dead body of drowning and the control group were dissected to observe the changes of lung, and the lung tissue was divided into two groups. One part was fixed with 4% paraformaldehyde polyformalin polyformaldehydede (PFA-PFA-neutral PBS(phosphate buffer salineine) solution for morphological examination, and one part of liquid nitrogen was frozen quickly.RNAs were extracted, qPCR(Real Time PCR and real-time fluorescence quantitative polymerase chain reaction (PCR) were used to screen stable internal reference genes and to detect AQP-5 quantitatively.Results compared with the control group, the lungs of the rats in each drowning group showed severe edema, rupture and hemorrhage to some extent, and obviously drowned fluid could be found in the lungs.The expression of ACTB (尾 -Actinin, 尾 -actin) was found to be the most stable internal reference gene, and the relative expression of AQP-5 in drowning rats was lower than that in control rats.Conclusion ACTB is the most stable internal reference gene, and the expression of AQP-5 mRNA is down-regulated at all drowning time points in rats.In this study, the expression of AQP-5 mRNA was positive in the drowning of rats, which was helpful for forensic drowning identification.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：D919

【相似文獻】

相關期刊論文 前10條

1 陳同度,張昌穎;素食大鼠的貧血現(xiàn)象[J];營養(yǎng)學報;1957年04期

2 陳偉強;趙善廣;;自制注射用大鼠固定裝置[J];上海實驗動物科學;1992年04期

3 肖柳英,林培英,馮昭明,張丹;不同周齡的SD大鼠生理、生化及體重的正常值測定[J];中藥新藥與臨床藥理;1996年03期

4 李淑云;簡易大鼠灌胃器的制作[J];錦州醫(yī)學院學報;2001年04期

5 楊明智,陳積圣;一種大鼠抓取與固定的新工具介紹[J];上海實驗動物科學;2001年03期

6 戴英,陸群;復方H_(505)對Wistar大鼠外周血的血液流變學指標的影響[J];中國血液流變學雜志;2001年01期

7 韋應波,孫喜慶,曹新生,姚永杰,馮岱雅,楊長斌;+Gz暴露時間對大鼠記憶功能和行為的影響[J];航天醫(yī)學與醫(yī)學工程;2003年01期

8 呂學軍,郭俊生,李敏,周利梅,張永娟;暈船大鼠體內(nèi)鐵含量的變化[J];中國職業(yè)醫(yī)學;2003年04期

9 湯仁仙,王迎偉,王慧,周峰;201A中藥合劑對大鼠抗腎小球基底膜腎炎病變的影響[J];徐州醫(yī)學院學報;2003年06期

10 孫同柱,付小兵,翁立新,梁雪梅,陳偉;介紹一種簡易的大鼠保定方法[J];上海實驗動物科學;2004年01期

相關會議論文 前10條

1 尹音;孫振宇;胡敏;李冬霞;;持續(xù)性高正加速度對大鼠顳頜關節(jié)損傷的作用[A];第八屆全國顳下頜關節(jié)病學及（牙合）學大會論文匯編[C];2011年

2 祝~=驤;iJ梊霞;洃克琴;崔素英;文允摪;，

本文編號：1760215