本文主要研究内容
作者董亚楠（2019）在《叶酸靶向的人参皂苷Rg5牛血清白蛋白纳米颗粒的制备及生物学研究》一文中研究指出：人参皂苷Rg5(Rg5)是从天然草本植物人参中提取的三萜类皂苷,对一些癌细胞具有不同程度的抑制作用,是最有效的抗癌药物之一。然而,Rg5的治疗潜力受其低的生物利用度,短的生物半衰期和差的靶向性等因素的限制。在纳米技术大背景下,研究工作者热衷于将纳米医药和肿瘤学相结合。纳米药物传送系统具有靶向性,可降解性和生物相容性,有望克服Rg5的缺点。因此,我们制备了叶酸(FA)靶向的Rg5负载的牛血清白蛋白(BSA)纳米颗粒,即FA-Rg5-BSA NPs。旨在增强Rg5药效同时,提高药物的靶向递送能力。基于此,本文主要的实验研究结果如下:(1)通过采用去溶剂化法制备Rg5-BSA NPs,利用酰胺化反应在Rg5-BSA NPs表面的活性氨基上修饰叶酸分子,制备新型的Rg5靶向药FA-Rg5-BSA NPs。采用单因素实验优化Rg5-BSA NPs和FA-Rg5-BSA NPs的制备工艺。最终优化的结果为:BSA浓度为15 mg/mL、BSA溶液pH值为9,水无水乙醇体积比1:3,Rg5与BSA摩尔比20:1,滴加乙醇速率为1.5-2.0 mL/min,搅拌转速600 r/min,搅拌时间24 h,叶酸与BSA反应的摩尔比为15:1,加入叶酸后反应时间6 h,离心转速10000 r/min。(2)制备出的FA-Rg5-BSA NPs水合直径为201.4 nm,PDI为0.081,且zeta电位是负值。呈小圆球形、外表光滑。载药量为12.64±4.02%,包封率为73.59±5.50%,叶酸偶联量为94.6μg/mg BSA NPs。FA-Rg5-BSA NPs水溶液在4℃下能维持8周左右的稳定性。在酸性(pH 5)条件,Rg5从颗粒中释放速度要快于在中性环境。(3)体外细胞MTT毒性实验证明,与Rg5-BSA NPs和Rg5相比,FA-Rg5-BSA NPs在MCF-7细胞中具有更高的抑制癌细胞增殖的能力,而在正常L929细胞中具通过有较高的细胞存活率。说明FA-Rg5-BSA NPs对不同种类的细胞有一定的选择性。通过Hoechst 33342及AO/EB荧光染色实验得出,在乳腺癌MCF-7细胞中FA-Rg5-BSA NPs比Rg5-BSA NPs和Rg5更能诱导细胞发生凋亡。观察FITC标记Rg5-BSA NPs和FA-Rg5-BSA NPs进入MCF-7细胞的摄取分布情况显示,细胞摄取FITC标记的FA-Rg5-BSA NPs的量要高于FITC标记的Rg5-BSA NPs,提前加入游离叶酸时,细胞摄取FITC标记的FA-Rg5-BSA NPs的量明显下降。这表明叶酸修饰的纳米颗粒可以通过叶酸受体介导的内吞途径促进细胞摄取。(4)在MCF-7异种移植肿瘤模型中体内药效研究表明,FA-Rg5-BSA NPs比Rg5-BSA NPs和Rg5更能抑制肿瘤生长,有较好的体内抗肿瘤活性。体内实时生物成像表明,FA-Rg5-BSA NPs比Rg5-BSA NPs有更好的肿瘤靶向作用。
Abstract
ren can zao gan Rg5(Rg5)shi cong tian ran cao ben zhi wu ren can zhong di qu de san tie lei zao gan ,dui yi xie ai xi bao ju you bu tong cheng du de yi zhi zuo yong ,shi zui you xiao de kang ai yao wu zhi yi 。ran er ,Rg5de zhi liao qian li shou ji di de sheng wu li yong du ,duan de sheng wu ban cui ji he cha de ba xiang xing deng yin su de xian zhi 。zai na mi ji shu da bei jing xia ,yan jiu gong zuo zhe re zhong yu jiang na mi yi yao he zhong liu xue xiang jie ge 。na mi yao wu chuan song ji tong ju you ba xiang xing ,ke jiang jie xing he sheng wu xiang rong xing ,you wang ke fu Rg5de que dian 。yin ci ,wo men zhi bei le xie suan (FA)ba xiang de Rg5fu zai de niu xie qing bai dan bai (BSA)na mi ke li ,ji FA-Rg5-BSA NPs。zhi zai zeng jiang Rg5yao xiao tong shi ,di gao yao wu de ba xiang di song neng li 。ji yu ci ,ben wen zhu yao de shi yan yan jiu jie guo ru xia :(1)tong guo cai yong qu rong ji hua fa zhi bei Rg5-BSA NPs,li yong xian an hua fan ying zai Rg5-BSA NPsbiao mian de huo xing an ji shang xiu shi xie suan fen zi ,zhi bei xin xing de Rg5ba xiang yao FA-Rg5-BSA NPs。cai yong chan yin su shi yan you hua Rg5-BSA NPshe FA-Rg5-BSA NPsde zhi bei gong yi 。zui zhong you hua de jie guo wei :BSAnong du wei 15 mg/mL、BSArong ye pHzhi wei 9,shui mo shui yi chun ti ji bi 1:3,Rg5yu BSAma er bi 20:1,di jia yi chun su lv wei 1.5-2.0 mL/min,jiao ban zhuai su 600 r/min,jiao ban shi jian 24 h,xie suan yu BSAfan ying de ma er bi wei 15:1,jia ru xie suan hou fan ying shi jian 6 h,li xin zhuai su 10000 r/min。(2)zhi bei chu de FA-Rg5-BSA NPsshui ge zhi jing wei 201.4 nm,PDIwei 0.081,ju zetadian wei shi fu zhi 。cheng xiao yuan qiu xing 、wai biao guang hua 。zai yao liang wei 12.64±4.02%,bao feng lv wei 73.59±5.50%,xie suan ou lian liang wei 94.6μg/mg BSA NPs。FA-Rg5-BSA NPsshui rong ye zai 4℃xia neng wei chi 8zhou zuo you de wen ding xing 。zai suan xing (pH 5)tiao jian ,Rg5cong ke li zhong shi fang su du yao kuai yu zai zhong xing huan jing 。(3)ti wai xi bao MTTdu xing shi yan zheng ming ,yu Rg5-BSA NPshe Rg5xiang bi ,FA-Rg5-BSA NPszai MCF-7xi bao zhong ju you geng gao de yi zhi ai xi bao zeng shi de neng li ,er zai zheng chang L929xi bao zhong ju tong guo you jiao gao de xi bao cun huo lv 。shui ming FA-Rg5-BSA NPsdui bu tong chong lei de xi bao you yi ding de shua ze xing 。tong guo Hoechst 33342ji AO/EBying guang ran se shi yan de chu ,zai ru xian ai MCF-7xi bao zhong FA-Rg5-BSA NPsbi Rg5-BSA NPshe Rg5geng neng you dao xi bao fa sheng diao wang 。guan cha FITCbiao ji Rg5-BSA NPshe FA-Rg5-BSA NPsjin ru MCF-7xi bao de she qu fen bu qing kuang xian shi ,xi bao she qu FITCbiao ji de FA-Rg5-BSA NPsde liang yao gao yu FITCbiao ji de Rg5-BSA NPs,di qian jia ru you li xie suan shi ,xi bao she qu FITCbiao ji de FA-Rg5-BSA NPsde liang ming xian xia jiang 。zhe biao ming xie suan xiu shi de na mi ke li ke yi tong guo xie suan shou ti jie dao de nei tun tu jing cu jin xi bao she qu 。(4)zai MCF-7yi chong yi zhi zhong liu mo xing zhong ti nei yao xiao yan jiu biao ming ,FA-Rg5-BSA NPsbi Rg5-BSA NPshe Rg5geng neng yi zhi zhong liu sheng chang ,you jiao hao de ti nei kang zhong liu huo xing 。ti nei shi shi sheng wu cheng xiang biao ming ,FA-Rg5-BSA NPsbi Rg5-BSA NPsyou geng hao de zhong liu ba xiang zuo yong 。
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论文作者分别是来自西北大学的董亚楠，发表于刊物西北大学2019-10-08论文，是一篇关于人参皂苷论文,牛血清白蛋白论文,纳米载药论文,靶向论文,抗肿瘤论文,西北大学2019-10-08论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自西北大学2019-10-08论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。