本文選題：丁香醛代謝物　切入點：輻射防護劑　出處：《安徽醫(yī)科大學》2013年碩士論文

【摘要】：在人類生存環(huán)境中，電離輻射時刻威脅著人們的健康，人們一直在尋求有效的防護藥物,目前廣泛應(yīng)用于臨床且效果滿意的輻射防護藥物并不多,尋找高效低毒活性高，不良反應(yīng)少的輻射防護劑變得日益重要。國內(nèi)外對輻射防護劑的研究都比較重視，除含硫化合物外，，細胞生長因子、天然藥物等以外，對細胞活素類，中草藥及復方、雌激素的抗輻射作用研究也較多。 天然化合物香蘭素，研究證實其具有抗氧化和抗誘變活性，清除電離輻射產(chǎn)生的自由基，抑制射線誘導的DNA和染色體損傷等，但其作用濃度高，活性相對較低。本實驗室篩選出輻射防護作用更好的香蘭素衍生物--丁香醛，已證實，丁香醛清除電離輻射產(chǎn)生的自由基的效果更好，可降低DNA損傷，提高DNA損傷修復能力以及輻射細胞存活率。丁香醛合成方便，理化性質(zhì)穩(wěn)定，具有開發(fā)成為新型輻射防護藥的價值。藥物代謝動力學發(fā)現(xiàn)，丁香醛進入動物體內(nèi)10～15min即可被機體吸收，達到血液循環(huán)的量明顯低于給藥劑量，在血漿里檢測到大量的丁香酸和少量的丁香醇，我們推測丁香醛進入體內(nèi)發(fā)揮輻射防護作用的是其主要代謝物--丁香酸或者兩種代謝物（丁香酸和丁香醇的混合物，以下簡稱混合物）協(xié)同發(fā)揮作用。本課題將從藥效學部分研究，利用細胞實驗觀察和比較丁香醛主要代謝物--丁香酸和混合物的輻射防護作用。利用動物實驗對丁香醛的主要代謝物--丁香酸的放射性氧化損傷和放射性造血損傷的防護作用進行研究，為丁香醛的進一步臨床應(yīng)用提供實驗依據(jù)。主要研究結(jié)果如下： 1.MTT方法觀察丁香醛代謝物對輻射后MEFs細胞效應(yīng)，結(jié)果顯示：和0μmol/L組相比，各加藥組的OD值明顯增加（p0.01），說明丁香醛代謝物能有效減輕輻射對MEF細胞的損傷。 2.MTT方法觀察丁香醛代謝物對照射后的MEFs細胞活力，結(jié)果顯示：和IR組相比，各加藥組的OD值明顯增加（p0.01），說明丁香醛代謝物能夠提高輻射后的MEF細胞活力。 3.細胞增殖實驗觀察丁香醛代謝物對照射后的MEFs細胞生長的影響，通過繪制細胞生長曲線顯示，和IR組相比，丁香醛代謝物能促進照射后MEFs細胞增殖。 4.細胞克隆形成實驗檢測丁香醛代謝物對照射后的MEFs細胞克隆形成率的影響，結(jié)果顯示：和IR組相比，各加藥組都能夠提高細胞克隆形成率，差異有統(tǒng)計學意義（p0.01），說丁香醛代謝物能提高照后MEFs細胞克隆形成率。 5．小鼠骨髓有核細胞計數(shù)，觀察丁香酸對小鼠骨髓造血損傷的作用，與IR組相比較，差異有統(tǒng)計學意義（p0.01），說明丁香酸對照后小鼠骨髓有核細胞有一定的輻射保護作用，能夠有效減輕小鼠骨髓造血損傷。 6.流式細胞術(shù)檢測小鼠骨髓有核細胞凋亡，結(jié)果顯示，與IR組相比較，差異有統(tǒng)計學意義（p0.01），說明丁香酸可降低輻射誘導的小鼠骨髓有核細胞凋亡。 7.照后取小鼠骨髓和脾臟組織，處理后光鏡下觀察各組動物的骨髓和脾臟的組織病理學變化，結(jié)果顯示：和IR組相比，丁香酸組小鼠的骨髓和脾臟組織損傷明顯減輕。 8.利用試劑盒檢測丁香酸對照后小鼠抗氧化損傷作用，與IR組相比較，丁香酸組在照后3d和6d，SOD活性的恢復都較顯著，差異有統(tǒng)計學意義(p0.01)，說明丁香酸可促進小鼠血清SOD活性的恢復，同時也降低了血清MDA的含量，與IR組比較有顯著的差異(p0.01)，丁香酸能夠增強細胞清除自由基的能力，減輕輻射誘發(fā)的膜脂質(zhì)過氧化損傷程度，減輕細胞的輻射損傷。 綜上所述，丁香醛代謝物對MEFs細胞具有輻射損傷防護作用，丁香醛主要代謝物--丁香酸對輻射小鼠也具有抗氧化損傷和造血損傷防護作用，為丁香醛的進一步應(yīng)用于臨床提供實驗依據(jù)。
[Abstract]:In the human survival environment, ionizing radiation, threatening people's health, people have been seeking the protection of effective drugs, radiation protection drugs currently used in clinical and the effect is not satisfactory, for high efficiency and low toxicity and high activity, less adverse reaction of the radiation protective agent becomes more and more important. The domestic and foreign research on radiation protection agent are more seriously, except sulphur compounds, cell growth factor, other than natural drugs, on cytokines, Chinese herbal medicine and compound, more research on anti radiation effect of estrogen.
Natural compounds vanillin, proved that it has antioxidant and antimutagenic activity, scavenging of ionizing radiation generated free radicals, inhibition of radiation-induced DNA and chromosomal damage, but the effect of high concentration of activity is relatively low. Our laboratory screened better radiation protective effect of vanillin derivatives -- syringaldehyde have confirmed that better effect of clove formaldehyde removal of ionizing radiation generated free radicals, can reduce DNA damage, improve the ability to repair DNA damage and radiation. The cell survival rate lilac aldehyde synthesis is convenient, stable physicochemical property, has developed as a new type of radiation protection drug found value. Pharmacokinetics of lilac aldehyde into animal body 10 ~ 15min can be absorbed by the body. Reach the blood circulation was significantly lower than the dosage, to a large number of syringic acid and a small amount of clove alcohol detection in the plasma, we speculate that syringaldehyde into play Radiation protection is the main metabolite -- syringic acid or two metabolites (Ding Xiangsuan and syringic alcohol mixture, hereinafter referred to as the synergistic mixture). This paper will study some research from the effect of radiation protective effect by cell experiment to observe and compare the main metabolites -- Ding Xiangsuan and lilac aldehyde mixture. The protective effect of the use of animal experiments the main metabolites of lilac aldehyde -- syringic acid radioactiveoxidative damage and radioactive hematopoietic injury were studied for further clinical application of syringaldehyde to provide experimental basis. The main results are as follows:
1.MTT method to observe the syringaldehyde metabolites on MEFs cells after radiation effect, the results show that compared with the 0 mol/L group, the OD values of each dosing group was significantly increased (P0.01), that syringaldehyde can effectively reduce the radiation of metabolites on MEF cell injury.
2.MTT method to observe the syringaldehyde metabolites on MEFs cell viability after irradiation, the results showed that compared with the IR group, the OD values of each dosing group was significantly increased (P0.01), that syringaldehyde metabolites can improve the viability of MEF cells after radiation.
3. cell proliferation experiments were conducted to observe the effects of syringal metabolites on the growth of MEFs cells after irradiation. By drawing cell growth curves, Ding Xiangquan metabolites could promote the proliferation of MEFs cells after irradiation compared with group IR.
4. cell clone formation experiment to detect the formation rate of syringaldehyde metabolites influence on MEFs cell clones after irradiation results show that compared with IR group, the experimental groups can improve cell colony formation rate, the difference was statistically significant (P0.01), said Ding Xiangquan metabolite MEFs after irradiation can improve the cell colony formation rate.
5. mice bone marrow nucleated cell count, observe the syringic acid injury of mouse bone marrow hematopoietic function, compared with the IR group, the difference was statistically significant (P0.01), indicates syringic acid in mouse bone marrow cells have some radiation protection effect, can effectively reduce the bone marrow hematopoietic damage in mice.
6. flow cytometry was used to detect the apoptosis of mouse bone marrow nucleated cells. The results showed that compared with group IR, the difference was statistically significant (P0.01), indicating that syringic acid could reduce the apoptosis of bone marrow nucleated cells induced by radiation.
The bone marrow and spleen tissues of mice after exposure to 7., changes of pathology were observed in animal bone marrow and spleen treatment under light microscope results showed that compared with the IR group, the bone marrow and spleen tissue injury mice syringic acid significantly reduced.
8. using the kit of syringic acid in mouse anti oxidative effect, compared with group IR, group 3D after irradiation of syringic acid and 6D, SOD activity recovery significantly, the difference was statistically significant (P0.01), indicates syringic acid can promote the activity of SOD in serum of mice recovered, while also reducing the content of the serum levels of MDA, there is a significant difference compared with IR group (P0.01), syringic acid can enhance the ability of the cells to eliminate free radicals, reducing lipid peroxidation induced by radiation damage, reduce the radiation injury of the cells.
In conclusion, Ding Xiangquan metabolites have protective effects on MEFs cells, and the main metabolite of Ding Xiangquan, syringic acid, has protective effects against oxidative damage and hematopoietic injury in radiation mice, providing experimental evidence for further application of syringaldehyde in clinic.

【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R818

【共引文獻】

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2 魏松宇;唐文偉;徐慧;梁瓊鶴;顧海斌;柴雪娥;郭斌;;小兒社區(qū)獲得性肺炎X線陽性預測模型的研究[J];東南大學學報(醫(yī)學版);2011年04期

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