本文選題：鈾 + WSC-DTPA納米粒��； 參考：《蘇州大學(xué)》2013年碩士論文

【摘要】：目的：通過細胞和動物實驗研究水溶性低分子量殼聚糖-二乙烯三氨五乙酸（WSC-DTPA）納米粒對硝酸鈾酰染毒的HK-2細胞和SD大鼠促排效果和防護作用。 方法：根據(jù)實驗室已建立的方法制備WSC-DTPA聚合物及其納米粒，再利用電鏡和粒徑儀檢測納米粒大小。細胞實驗：采用MTT法檢測WSC-DTPA納米粒對硝酸鈾酰染毒的正常腎近曲小管上皮細胞(HK-2)存活率的影響；流式細胞儀PI單染法檢測WSC-DTPA納米粒作用硝酸鈾酰染毒的HK-2細胞的細胞周期分布份額變化；流式細胞儀熒光法檢測WSC-DTPA納米粒作用硝酸鈾酰染毒的HK-2細胞內(nèi)的ROS含量；ICP-AES檢測經(jīng)過WSC-DTPA納米粒作用硝酸鈾酰染毒后的HK-2細胞內(nèi)鈾含量。動物實驗：采用腹腔注射硝酸鈾酰（0.5mgU/kg）方式對SD大鼠進行染毒，即刻尾靜脈注射92.7%WSC-DTPA納米粒、1.59%WSC-DTPA納米粒、WSC納米粒及促排靈（DTPA-Na_3Ca），給藥劑量均為60mg/kg。ICP-AES測量SD大鼠第1天、第2天、第3天的尿和糞中鈾的排出量，以及給藥3天后肝、脾、腎和骨中鈾蓄積量；采用腹腔注射硝酸鈾酰（2.0mgU/kg）方式對SD大鼠進行染毒，相同方式處理，5天后，取肝、腎、脾做病理切片，骨髓作骨髓細胞涂片，查看病理損傷程度。評價WSC-DTPA納米粒的對內(nèi)污染鈾的促排和輻射防護效果。 結(jié)果：制備出的WSC-DTPA納米粒經(jīng)檢測，符合實驗要求。細胞實驗，MTT法結(jié)果表明，鈾濃度在12.5μg/mL左右HK-2細胞的存活率接近50%。MTT實驗發(fā)現(xiàn)DTPA-CaNa_3和1.59%WSC-DTPA納米粒不能提高12.5μg/mL鈾染毒HK-2細胞的存活率，而92.7%WSC-DTPA納米粒和WSC納米粒能明顯提高細胞的存活率（P㩳0.05），并且存活率隨著給藥濃度的增加而提高。流式細胞PI單染法結(jié)果表明，12.5μg/mL鈾染毒細胞出現(xiàn)G0/G1期阻滯，該期的DNA含量顯著增高（P㩳0.05），而S期含量明顯減少，G2/M期的含量大致不變，維持在15%左右；DTPA-CaNa_3和1.59%WSC-DTPA納米粒不能改善鈾染毒造成的G0/G1期阻滯現(xiàn)象，而92.7%WSC-DTPA納米粒和WSC納米粒能顯著地緩減G0/G1期阻滯。細胞促排實驗結(jié)果表明，與單純?nèi)径荆?.13μg/mL）組相比， DTPA-CaNa_3、1.59%WSC-DTPA納米粒、92.7%WSC-DTPA納米粒和WSC納米粒均有促排效果（P㩳0.05），，其中1.59%WSC-DTPA納米粒的促排效果最佳，不同時間給藥實驗發(fā)現(xiàn)立刻給藥的促排效果相對較好。細胞內(nèi)ROS含量實驗結(jié)果表明，鈾染毒可造成細胞內(nèi)的ROS含量明顯增加（P㩳0.05），92.7%WSC-DTPA納米粒和WSC納米粒能有效地減少細胞內(nèi)的ROS含量（P㩳0.05），而1.59%WSC-DTPA納米粒和DTPA-CaNa_3不能減少細胞內(nèi)的ROS含量�？梢�92.7%WSC-DTPA納米粒和WSC納米粒具有抗輻射效果。動物實驗結(jié)果顯示，在0.5μg/ml鈾染毒實驗中，92.7%WSC-DTPA納米粒、1.59%WSC-DTPA和DTPA-CaNa_3均有促排效果，且1.59%WSC-DTPA納米粒促排效果明顯優(yōu)于DTPA-CaNa_3組，統(tǒng)計學(xué)上有顯著性差異（P㩳0.05）；給藥組的主要組織鈾蓄積量明顯低于單純?nèi)径窘M，有顯著性統(tǒng)計學(xué)差異（P㩳0.05）。在2.0μg/ml鈾染毒實驗中，92.7%WSC-DTPA納米粒和WSC納米粒的主要組織鈾蓄積量明顯低于DTPA-CaNa_3和1.59%WSC-DTPA納米粒，統(tǒng)計學(xué)上有顯著性差異（P㩳0.05）；病理組織切片顯示92.7%WSC-DTPA納米粒和WSC納米粒對肝、腎有保護作用，損傷明顯要比其他藥物組輕�？梢娫谳^高劑量鈾染毒的情況下對組織的防護作用相對比較重要的。骨髓圖片的細胞分類未見有明顯異常。 結(jié)論：WSC-DTPA納米粒對內(nèi)污染鈾有較好促排效果，其中1.59%WSC-DTPA納米粒的促排效果最好；92.7%WSC-DTPA納米粒不僅有促排作用，而且對鈾染毒的大鼠的肝、腎組織有輻射防護作用。
[Abstract]:Objective: To study the effect and protective effect of water-soluble low molecular weight chitosan, two ethylene three ammonia five acetic acid (WSC-DTPA) nanoparticles, on HK-2 cells and SD rats infected with uranyl nitrate.
Methods: the WSC-DTPA polymer and its nanoparticles were prepared according to the method established in the laboratory. The size of the nanoparticles was detected by electron microscopy and particle size analyzer. The effect of WSC-DTPA nanoparticles on the survival rate of the normal renal proximal tubular epithelial cells (HK-2) was detected by the MTT method. The flow cytometry PI single staining method was used to detect WSC. The changes in the cell cycle distribution of the HK-2 cells infected with -DTPA nanoparticles; the flow cytometer fluorescence assay was used to detect the ROS content in HK-2 cells infected with WSC-DTPA nanoparticles; ICP-AES was used to detect the content of uranium in HK-2 cells infected by WSC-DTPA nanoparticles in the presence of uranyl nitrate. Animal experiments: SD rats were poisoned by intraperitoneal injection of uranyl nitrate (0.5mgU/kg). 92.7%WSC-DTPA nanoparticles, 1.59%WSC-DTPA nanoparticles, WSC nanoparticles and DTPA-Na_3Ca (DTPA-Na_3Ca) were injected into the tail vein. The dosage of the drug was measured by 60mg/kg.ICP-AES in SD rats for first days, second days, third days of urine and excretion of uranium, and 3 days after the administration of the liver and spleen. The amount of uranium accumulated in the kidney and bone; the SD rats were treated by intraperitoneal injection of uranyl nitrate (2.0mgU/kg) and treated in the same way. After 5 days, the liver, kidney and spleen were taken to make pathological sections. The bone marrow smear was used as the bone marrow cell smear to examine the degree of pathological damage. The effect of WSC-DTPA nanoparticles on internal pollution of uranium and radiation protection were evaluated.
Results: the prepared WSC-DTPA nanoparticles were tested and met the experimental requirements. Cell experiments and MTT results showed that the survival rate of uranium concentration at about 12.5 u g/mL near 50%.MTT showed that DTPA-CaNa_3 and 1.59%WSC-DTPA nanoparticles could not increase the survival rate of HK-2 fine cells infected by 12.5 u g/mL uranium, while 92.7%WSC-DTPA nanoparticles and WSC Na The rice grain survival rate (P? 0.05) was significantly increased, and the survival rate increased with the increase of drug concentration. The results of PI single staining in flow cytometry showed that the G0/G1 phase block appeared in the 12.5 u g/mL infected cells, and the DNA content in this period increased significantly (P? 0.05), while the content of S phase decreased significantly, and the content of G2/M phase remained approximately 15%; DT; DT PA-CaNa_3 and 1.59%WSC-DTPA nanoparticles did not improve the G0/G1 phase blocking phenomenon caused by uranium exposure, while 92.7%WSC-DTPA nanoparticles and WSC nanoparticles could significantly reduce the G0/G1 phase block. The results of cell promotion experiments showed that DTPA-CaNa_3,1.59% WSC-DTPA nanoparticles, 92.7%WSC-DTPA nanoparticles and WSC nanoparticles were compared with those of the group of pure (3.13 mu g/mL). The effect of P? 0.05 was the best, and the effect of 1.59%WSC-DTPA nanoparticles was the best. The effect of immediate delivery was better. The results of intracellular ROS content showed that the content of ROS in the cells increased significantly (P? 0.05), and the 92.7%WSC-DTPA nanoparticles and WSC nanoparticles could be effectively reduced. The intracellular ROS content (P? 0.05) and 1.59%WSC-DTPA nanoparticles and DTPA-CaNa_3 did not reduce the ROS content in the cells. The 92.7%WSC-DTPA nanoparticles and WSC nanoparticles had radiation resistance. The experimental results showed that the 92.7%WSC-DTPA nanoparticles, 1.59%WSC-DTPA and DTPA-CaNa_3 had the effect of stimulating, and 1.5 in the 0.5 u g/ml uranium exposure test. The effect of 9%WSC-DTPA nanoparticles was significantly better than that in the DTPA-CaNa_3 group (P? 0.05); the main tissue uranium accumulation of the drug group was significantly lower than that of the pure exposure group (P? 0.05). In the 2 UU g/ml uranium exposure experiment, the main tissue uranium accumulation of 92.7% WSC-DTPA nanoparticles and WSC nanoparticles Significantly lower than DTPA-CaNa_3 and 1.59%WSC-DTPA nanoparticles, statistically significant differences (P? 0.05); pathological tissue sections show that 92.7%WSC-DTPA nanoparticles and WSC nanoparticles have protective effects on the liver and kidney, and the damage is significantly lighter than that of other drug groups. There were no obvious abnormalities in the cell classification of bone marrow images.
Conclusion: WSC-DTPA nanoparticles have better effect on promoting and discharging of internal contaminated uranium, and the effect of 1.59%WSC-DTPA nanoparticles is the best, and 92.7%WSC-DTPA nanoparticles not only have the effect of stimulating, but also have radiation protection against the liver and kidney of rats infected with uranium.

【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R818.05

【參考文獻】

相關(guān)期刊論文 前10條

1 劉維俊;高分子殼聚糖對微量金屬離子的螯合作用研究[J];環(huán)境科學(xué)與技術(shù);2003年03期

2 陳紅紅,胡昱興,王英華,金美英,羅梅初,孫梅貞;鄰苯二酚氨羧酸螯合劑對放射性釷的促排效果比較[J];輻射防護;2003年04期

3 劉濤;張貴鋒;周衛(wèi)斌;蘇志國;;凝膠過濾色譜-多角度激光散射法測定琥珀酰明膠自由氨基修飾度[J];分析化學(xué);2007年01期

4 劉蘊瑩;周效旺;萬建美;胡丹飛;張友九;;WSC-DTPA納米粒輻射防護作用的初步研究[J];輻射研究與輻射工藝學(xué)報;2012年05期

5 高禮;;殼聚糖應(yīng)用于水處理的化學(xué)基礎(chǔ)[J];水科學(xué)與工程技術(shù);2008年S2期

6 王麗麗;林強;;殼聚糖印跡分子吸附性能的研究進展[J];化學(xué)世界;2010年08期

7 曹珍山,朱茂祥,楊陟華,潘秀頡,李元敏;大鼠吸入貧鈾氣溶膠后主要器官的病理損傷特點[J];中國輻射衛(wèi)生;2005年02期

8 張斌;丁新民;段蘊鈾;;貧鈾對機體健康影響的研究進展[J];中國預(yù)防醫(yī)學(xué)雜志;2011年05期

9 匡長春,何文,羅順德,宋金春;去甲斑蝥素殼聚糖納米粒在小鼠體內(nèi)分布及肝靶向作用[J];中國醫(yī)院藥學(xué)雜志;2005年06期

10 羅梅初,陳紅紅;放射性核素內(nèi)污染促排藥物研究[J];中華放射醫(yī)學(xué)與防護雜志;2000年01期

本文編號：1849449