人外周血淋巴細(xì)胞pig3和gdf15基因表達(dá)變化作為輻射生物劑量計(jì)的研究
發(fā)布時間：2019-07-03 16:20
【摘要】：目的： 隨著人類核活動的日益增多,核與輻射的意外暴露以及核恐怖事件發(fā)生的可能性也較以往有所增大。在大人群受到照射的情況下,傳統(tǒng)的生物劑量估算方法雙著絲粒和著絲粒環(huán)畸變分析不能滿足快速生物劑量估算的需求,因此需要建立一種快速、高通量的生物標(biāo)志物,以應(yīng)對大批量人群輻射暴露情況下的生物劑量估算。因此本研究試圖使用目前較為先進(jìn)的Real-time PCR、ELISA等技術(shù),對電離輻射誘導(dǎo)的人外周血淋巴細(xì)胞中pig3和gdfl5基因表達(dá)進(jìn)行定量測定,并對其變化規(guī)律進(jìn)行分析,這將有助于電離輻射的分子水平生物效應(yīng)及其機(jī)制的闡明,為輻射損傷研究提供新的生物標(biāo)志物,也將為發(fā)現(xiàn)更加有效、快速、高通量的生物劑量計(jì)積累理論依據(jù)。 方法： 1、在知情同意情況下,收集73名正常人外周血樣品。利用Taqman MGB探針法絕對定量分析正常人外周血淋巴細(xì)胞中pig3和gdf15基因表達(dá)水平的本底值。 2、利用Taqman MGB探針法和ELISA技術(shù)分析受照射人外周血淋巴細(xì)胞中pig3和gdf15基因在照后多個時間點(diǎn)(4h、8h、12h、24h、48h和72h)轉(zhuǎn)錄水平和翻譯水平的劑量效應(yīng)關(guān)系,劑量為0、1、3、5和8Gy,最終通過統(tǒng)計(jì)學(xué)方法擬合劑量效應(yīng)曲線；并確定時間響應(yīng)范圍。 3、通過分析受到不同劑量(OGy、2Gy、4Gy.6Gy)60Co γ射線照射后SD大鼠離體和整體外周血中g(shù)df15基因表達(dá)變化,驗(yàn)證兩種基因整體與離體效應(yīng)是否一致,探索兩種基因作為輻射生物劑量計(jì)應(yīng)用的準(zhǔn)確性。 結(jié)果： 1、在轉(zhuǎn)錄水平,正常人外周血pig3和gdf15基因表達(dá)的本底拷貝數(shù)在不同性別和不同年齡組之間沒有統(tǒng)計(jì)學(xué)差異。 2.0～8Gy60Coγ射線照射后8h～48h人外周血淋巴細(xì)胞pig3和gdf15基因表達(dá)水平均呈現(xiàn)良好的劑量效應(yīng)關(guān)系,并可擬合成為R2值較高的直線；照射后72h出現(xiàn)了隨劑量增加,pig3和gdf15基因表達(dá)先上調(diào)后下調(diào)的趨勢。 3、在翻譯水平,0～8Gy60Coγ射線照射后48h和72h,在一定的劑量范圍內(nèi),人外周血淋巴細(xì)胞PIG3蛋白表達(dá)呈現(xiàn)一定的劑量效應(yīng)關(guān)系,可以擬合成為R2值較高的直線；其他時間點(diǎn)沒有明顯的劑量效應(yīng)關(guān)系。 結(jié)論： 1、止常人外周血pig3和gdf15基因mRNA本底水平不受供血者性別和年齡的影響,這是兩種基因表達(dá)水平成為輻射生物劑量計(jì)的基礎(chǔ)。 2、在mRNA水平,離體照射的人外周血pig3和gdf15基因表達(dá)在0～8Gy劑量照射后8h～48h均表現(xiàn)出了良好的劑量效應(yīng)關(guān)系,證實(shí)了兩種基因表達(dá)變化具備了成為生物劑量計(jì)的基本條件。 3、PIG3蛋白水平在0～5Gy劑量照射后48h和72h具有良好的劑量效應(yīng)關(guān)系。
[Abstract]:Purpose: With the increasing number of human nuclear activities, the accidental exposure of nuclear and radiation, as well as the possibility of nuclear terrorism, has also increased in the past Large. In the case of large population exposure, the traditional biological dose estimation method, the double centromere and the centromere ring distortion analysis, cannot meet the requirement of rapid biological dose estimation, therefore, a rapid and high-flux biological marker needs to be established. To deal with the estimation of the biological dose in the case of exposure to radiation in a large number of people In this study, we tried to measure the expression of pg3 and gdfl5 in human peripheral blood lymphocytes induced by ionizing radiation by using the technology of real-time PCR, ELISA and so on. This will help the molecular level biological effect of ionizing radiation and its mechanism, provide a new biological marker for the research of radiation damage, and will also find a more effective, rapid and high-flux biological dosimeter accumulation theory. It was reported. Method:1. Under the condition of informed consent,73 normal persons were collected. quantitative analysis of the expression of pg3 and gdf15 in peripheral blood lymphocytes from peripheral blood samples by using the Taqman MGB probe method the level of the pg3 and gdf15 genes in the peripheral blood lymphocytes of the irradiated human were analyzed by using the Taqman MGB probe method and the ELISA technique, and the dose effect relationship between the transcription level and the translation level of the human peripheral blood lymphocytes after the irradiation (4h, 8h, 12h, 24h, 48h and 72h) was analyzed, and the dose was 0,1, 3,5 and 8 Gy, and the dose response curves were finally fitted by a statistical method; and 3. By analyzing the expression of gdf15 gene in the isolated and whole peripheral blood of SD rats exposed to different doses (OGy, 2Gy, 4Gy. 6Gy), the expression of gdf15 gene in the isolated and whole peripheral blood of SD rats was changed. dosimeter The results were as follows:1. The background copy number of the expression of the pg3 and gdf15 genes in the peripheral blood of the normal human is different from the different sex and not in the normal human. There was no statistical difference between the same age groups. The expression levels of the pg3 and gdf15 gene in the peripheral blood lymphocytes of the human peripheral blood lymphocytes from 8 h to 48 hours after the irradiation of 2.0 to 8 Gy 60Co showed a good dose-effect relationship and can be fitted into a straight line with a higher R2 value. The dose-increasing, the pige3 and the gdf occurred at 72 h after the irradiation. The expression of PIG3 protein in peripheral blood of human peripheral blood showed a dose-effect relationship at 48 h and 72 h after the level of translation, and the expression of PIG3 protein in human peripheral blood lymphocytes could be fit into a straight line with high R2 value. ; others Conclusion:1. The level of the normal human peripheral blood pige3 and gdf15 gene mRNA is not affected by the sex and age of the donor. The expression level of the two genes was the basis of the radiation biological dosimeter.2. The expression of the human peripheral blood pige3 and gdf15 in the in-vitro irradiation at the mRNA level showed a good dose-effect relationship between 8 and 48 hours after the dose of 0-8Gy. The expression of the two genes is the basic condition of the biological dosimeter.3, the level of the PIG3 protein is 0-5.
【學(xué)位授予單位】：中國疾病預(yù)防控制中心
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R142
本文編號：2509519
[Abstract]:Purpose: With the increasing number of human nuclear activities, the accidental exposure of nuclear and radiation, as well as the possibility of nuclear terrorism, has also increased in the past Large. In the case of large population exposure, the traditional biological dose estimation method, the double centromere and the centromere ring distortion analysis, cannot meet the requirement of rapid biological dose estimation, therefore, a rapid and high-flux biological marker needs to be established. To deal with the estimation of the biological dose in the case of exposure to radiation in a large number of people In this study, we tried to measure the expression of pg3 and gdfl5 in human peripheral blood lymphocytes induced by ionizing radiation by using the technology of real-time PCR, ELISA and so on. This will help the molecular level biological effect of ionizing radiation and its mechanism, provide a new biological marker for the research of radiation damage, and will also find a more effective, rapid and high-flux biological dosimeter accumulation theory. It was reported. Method:1. Under the condition of informed consent,73 normal persons were collected. quantitative analysis of the expression of pg3 and gdf15 in peripheral blood lymphocytes from peripheral blood samples by using the Taqman MGB probe method the level of the pg3 and gdf15 genes in the peripheral blood lymphocytes of the irradiated human were analyzed by using the Taqman MGB probe method and the ELISA technique, and the dose effect relationship between the transcription level and the translation level of the human peripheral blood lymphocytes after the irradiation (4h, 8h, 12h, 24h, 48h and 72h) was analyzed, and the dose was 0,1, 3,5 and 8 Gy, and the dose response curves were finally fitted by a statistical method; and 3. By analyzing the expression of gdf15 gene in the isolated and whole peripheral blood of SD rats exposed to different doses (OGy, 2Gy, 4Gy. 6Gy), the expression of gdf15 gene in the isolated and whole peripheral blood of SD rats was changed. dosimeter The results were as follows:1. The background copy number of the expression of the pg3 and gdf15 genes in the peripheral blood of the normal human is different from the different sex and not in the normal human. There was no statistical difference between the same age groups. The expression levels of the pg3 and gdf15 gene in the peripheral blood lymphocytes of the human peripheral blood lymphocytes from 8 h to 48 hours after the irradiation of 2.0 to 8 Gy 60Co showed a good dose-effect relationship and can be fitted into a straight line with a higher R2 value. The dose-increasing, the pige3 and the gdf occurred at 72 h after the irradiation. The expression of PIG3 protein in peripheral blood of human peripheral blood showed a dose-effect relationship at 48 h and 72 h after the level of translation, and the expression of PIG3 protein in human peripheral blood lymphocytes could be fit into a straight line with high R2 value. ; others Conclusion:1. The level of the normal human peripheral blood pige3 and gdf15 gene mRNA is not affected by the sex and age of the donor. The expression level of the two genes was the basis of the radiation biological dosimeter.2. The expression of the human peripheral blood pige3 and gdf15 in the in-vitro irradiation at the mRNA level showed a good dose-effect relationship between 8 and 48 hours after the dose of 0-8Gy. The expression of the two genes is the basic condition of the biological dosimeter.3, the level of the PIG3 protein is 0-5.
【學(xué)位授予單位】：中國疾病預(yù)防控制中心
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R142
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