本文選題：Y_2O_3NSs + 溶解性��； 參考：《河北大學(xué)》2017年碩士論文

【摘要】：隨著納米技術(shù)的發(fā)展,稀土納米材料由于其獨特的光、電、磁的性質(zhì)已廣泛的應(yīng)用于生物醫(yī)學(xué)領(lǐng)域。氧化釔納米材料被大量用作熒光探針、藥物載體、骨支架材料等,文獻報道稱稀土納米材料易于在骨中蓄積且不易排出,然而氧化釔納米材料(Y_2O_3 NPs)對骨代謝的影響還不是很清楚。本文研究了氧化釔納米球(Y_2O_3 NSs)與骨髓間充質(zhì)干細胞(BMSCs)之間的相互作用。水熱法制備Y_2O_3 NSs,通過SEM、TEM、DLS等手段進行表征。MTT法等等方法檢測Y_2O_3 NSs對細胞活力的影響。在細胞水平上,Annexin V/PI雙染法,攝取機制研究,定位,組織蛋白酶的釋放,Bid和Bax的表達,細胞色素C(Cyt C)的釋放,Caspase 3的激活等方法研究了Y_2O_3 NSs對BMSCs溶酶體-線粒體依賴的凋亡通路的影響。在溶酶體模擬液中合成了磷酸根包覆的氧化釔(Y_2O_3@YPO4),采用TEM、Zeta、FTIR等方法進行表征,Y_2O_3 NSs及Y_2O_3@YPO4對組織蛋白酶的釋放及Caspase 3表達的比較,探討了Y_2O_3 NSs在溶酶體內(nèi)的物種變化及Y_2O_3@YPO4安全性的設(shè)計思路。在動物水平上,通過HE染色,末端標(biāo)記法(TUNEL)、Caspase 3的表達等手段檢測了Y_2O_3 NSs對ICR小鼠的體內(nèi)骨代謝的影響。結(jié)果表明,合成了粒徑約為160 nm的Y_2O_3 NSs,MTT法表明Y_2O_3 NSs對BMSCs具有濃度、時間依賴的細胞毒性。Y_2O_3NSs通過大胞飲的形式進入細胞,定位于溶酶體,并且在溶酶體內(nèi)溶解,釋放出的Y3+與溶酶體內(nèi)的磷酸根絡(luò)合,形成蜘蛛網(wǎng)狀結(jié)構(gòu),破壞了溶酶體的穩(wěn)態(tài),最終通過Caspase3依賴的方式誘導(dǎo)了細胞凋亡,而Y_2O_3@YPO4減少了組織蛋白酶的釋放,降低了Caspase3的表達以及Cyt C的釋放,體內(nèi)實驗結(jié)果表明Y_2O_3 NSs誘導(dǎo)部分肝細胞壞死,肺泡的破裂,骨小梁變細,減少,并且激活骨組織中Caspase 3的表達,而Y_2O_3.@YPO4對骨組織沒有明顯的毒性。因此,通過Y_2O_3 NSs在體內(nèi)外的毒性評價表明Y_2O_3 NSs在溶酶體內(nèi)的溶解是引起細胞凋亡的關(guān)鍵,Y_2O_3@YPO4為稀土氧化物對生物體內(nèi)的安全性設(shè)計提供了一種新思路。
[Abstract]:With the development of nanotechnology, rare earth nanomaterials have been widely used in biomedical fields due to their unique optical, electrical and magnetic properties. Yttrium oxide nanomaterials are widely used as fluorescent probes, drug carriers, bone scaffolds and so on. However, the effect of Y _ S _ 2O _ 3 NPs on bone metabolism is unclear. The interaction between yttrium oxide nanospheres (Ys) and bone marrow mesenchymal stem cells (BMSCs) has been studied. Ys _ 2O _ 3 NSs were prepared by hydrothermal method. The effects of Ys _ 2O _ 3 NSs on cell viability were detected by means of SEMT-TEML-DLS and other methods, such as MTT method. At the cellular level, Annexin V / Pi double staining, uptake mechanism studies, localization, cathepsin release and expression of bid and Bax, The effects of Y _ 2O _ 3 NSs on lysosome-mitochondrial dependent apoptosis pathway of BMSCs were studied by the activation of Caspase 3 by the release of cytochrome Cyt C, and the activation of Caspase 3. Yttrium phosphate coated yttrium oxide YPO4 was synthesized in lysosomal mimic solution. The expression of cathepsin and expression of Caspase 3 were characterized by means of TEMN ZetaI FTIR and the comparison of the release of cathepsin and the expression of Caspase 3 in Y2O3 / YPO4 and Y2O3 / YPO4, respectively. The species changes in lysosomes of Ys _ 2O _ 3NSs and the safety of Ys _ 2O _ 3NSs in lysosomes were discussed. At the animal level, the effects of Y _ 2O _ 3 NSs on bone metabolism in ICR mice were detected by HE staining and end labeling method. The results showed that Ys _ 2O _ 3 NSs with a diameter of about 160nm were synthesized. The results showed that Ys _ 2O _ 3NSs had concentration and time-dependent cytotoxicity to BMSCs. Ys _ 2O _ 3NSs entered the cells through the form of large cell drink, located in lysosomes, and dissolved in lysosomes. The released Y3 complex with phosphates in lysosomes to form spider reticular structure, destroy the steady-state of lysosome, induce cell apoptosis by Caspase3-dependent manner, and Y2O3YPO4 reduce the release of cathepsin, and Y2O3-dependent YPO4 reduces the release of cathepsin, and Y2O3YPO4 reduces the release of cathepsin. The expression of Caspase3 and the release of Cyt C were decreased. The results of in vivo experiments showed that Y2O3 NSs induced partial hepatocyte necrosis, alveolar rupture, bone trabecula thinning, and activation of Caspase 3 expression in bone tissue. But Ys _ 2 O _ 3. No obvious toxicity to bone tissue. Therefore, the evaluation of the toxicity of Y2O3NSs in vivo and in vitro shows that the dissolution of Y _ 2O _ 3NSs in the lysosome is the key to the apoptosis of the cells. Y2O3OP-YPO4 provides a new idea for the safety design of rare earth oxides to organisms.
【學(xué)位授予單位】：河北大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R99;TB383.1

【參考文獻】

相關(guān)期刊論文 前1條

1 李媛;王小慧;莊遠;張華欣;閻少多;詹林盛;;不同尺寸、形貌金納米粒子小鼠體內(nèi)急性毒性研究[J];軍事醫(yī)學(xué);2013年06期

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本文編號：1981662