本文主要研究内容
作者张朵朵（2019）在《金纳米荷载白藜芦醇的制备表征及抗肝癌作用研究》一文中研究指出：癌症作为一个日益严重的全球健康问题,每年都影响着数百万人。其中,肝癌是世界上第五大常见恶性肿瘤,也是导致死亡的第二大原因。在临床医生和科学家的共同努力下,肝癌的总体生存率得以延长。然而,在治疗过程中,放疗和化疗药物造成的的不良反应,大大降低了患者的生存率。因此,迫切需要研究新的药物来提高我们诊断、预防和治疗肝癌的能力。纳米粒子作为运载工具为医学诊断和治疗提供了巨大的潜力。银纳米和金纳米具有良好的生物相容性、低毒性和功能多样性,并且在生物医学领域有着广泛的应用。由于金纳米颗粒表面积大,能被多种治疗药物包覆,因此在药物传递系统中具有广阔的应用前景。最近的研究表明,金纳米颗粒已被报道应用于抗肿瘤治疗,包括癌症抗原和佐剂的传递。根据金纳米颗粒的特性,本实验室猜想纳米技术是否可以应用于增强白藜芦醇的抗肿瘤作用。白藜芦醇是一种天然的化合物,存在于浆果、坚果、虎杖和葡萄中。近年来,大量的研究表明白藜芦醇具有多种药理学效应,包括潜在的抗癌、抗炎、心脏保护和神经保护活性。但是,白藜芦醇难溶于水和生物利用度低的特点大大限制了它的应用。目前,有很多研究者通过对白藜芦醇进行修饰来改善它的缺点。基于这些发现,本课题在体外和体内进行了一系列研究,以探讨金纳米颗粒是否可用于增强Res的抗肿瘤作用。本实验利用白藜芦醇自身具备的还原性,以白藜芦醇和氯金酸溶液为原料,在37℃、质量比为1:2、反应时间为120 min的条件下,一步法制备粒径均一、分散性良好的金纳米荷载白藜芦醇。紫外光谱结果表明金纳米荷载白藜芦醇的等离子共振吸收峰在539 nm处出现。透射电镜实验结果表明通过调节反应温度、时间和质量比可以得到粒径在39 nm的金纳米荷载白藜芦醇。另外,红外光谱实验数据表明金与白藜芦醇酚羟基结构中氧的协同作用,使白藜芦醇包裹在金纳米粒子表面,白藜芦醇中的碳碳双键和羟基得到了保留,并且已有大量研究表明碳碳双键和羟基在白藜芦醇发挥很多药理作用中起到重要的作用。因此,我们预测金纳米荷载白藜芦醇可能具有白藜芦醇的部分药理作用和金纳米颗粒的优良特性,在抗肿瘤方面具有良好的应用前景。MTT实验检测白藜芦醇和金纳米荷载白藜芦醇对Hepg2细胞生长的影响;AO染色法检测药物对肝癌细胞形态的影响;流式细胞仪检测肝癌细胞的凋亡情况;透射电镜检测金纳米荷载白藜芦醇在Hepg2细胞内的分布情况;蛋白印迹免疫法（Western Blot）检测Caspase家族和PI3K/Akt等相关凋亡蛋白的变化;MTT实验检测白藜芦醇和金纳米荷载白藜芦醇对正常肝细胞L02和肾小球系膜GMCs细胞的毒性作用。在以上实验的基础上,用BALB/c裸鼠构建人肝癌Hepg2细胞荷瘤模型,持续给予药物42天,处死荷瘤裸鼠,剥离瘤体,HE染色观察肿瘤组织的形态学变化;Tunel染色检测肿瘤组织中细胞凋亡的情况;免疫组化法检测瘤体中血管生长因子（VEGF）的表达。摘取动物的心脏、肝脏、脾脏、肺脏和肾脏,HE切片染色观察受试物对上述器官组织形态学改变的影响,同时用MTT法检测药物对L02和GMCs细胞的毒性。MTT结果表明白藜芦醇和金纳米荷载白藜芦醇对Hepg2细胞均具有明显的抑制作用,并且金纳米荷载白藜芦醇的抑制作用更显著。白藜芦醇和金纳米荷载白藜芦醇处理Hepg2细胞24 h后,半数抑制浓度(IC50)分别为27.74g/ml和3.84g/ml,为了进一步检测白藜芦醇和金纳米荷载白藜芦醇对L02细胞和GMCs细胞的毒性作用,分别用白藜芦醇和金纳米荷载白藜芦醇处理24 h,金纳米荷载白藜芦醇对L02细胞和GMCs细胞的抑制率依次为35%和10%,而白藜芦醇组的抑制率依次为83%和63%。AO染色结果表明两种药物均能够促进细胞凋亡,导致细胞发生形态学改变,给药后Hepg2细胞的细胞核膜皱缩,细胞体积变小而呈绿色的荧光,凋亡细胞呈碎片状。随着给药浓度的递增,其凋亡细胞数也相应增加。透射电镜结果表明功能化的金纳米荷载白藜芦醇在细胞内大量积累,并通过线粒体途径诱导细胞凋亡。流式细胞仪结果也表明细胞的凋亡有显著的剂量依赖性。WB检测相关蛋白发现,金纳米荷载白藜芦醇可显著上调激活的caspase-3、caspase-8、caspase-9与bax和下调PI3K与Akt的表达。白藜芦醇对上述蛋白的调节也呈相同趋势,但调节作用弱于金纳米荷载白藜芦醇。荷瘤实验结束后,剥离瘤体并称重。发现白藜芦醇和金纳米荷载白藜芦醇对瘤体的生长均有较强抑制作用。白藜芦醇组和金纳米荷载白藜芦醇组中裸鼠移植瘤体的体积和重量均明显的降低,对瘤重的抑制率分别为35.5%和57.2%。荷瘤组织经HE染色后,结果显示,模型组肿瘤组织中的细胞排列整齐,细胞形态完整;给药组的细胞随着浓度的递增细胞数量逐渐减少,细胞核膜皱缩,呈均匀蓝色,相较于白藜芦醇组,金纳米荷载白藜芦醇组诱导细胞凋亡的效果更加显著;HE染色结果表明,发现金纳米荷载白藜芦醇对裸鼠的心脏、肝脏、脾脏、肺脏和肾脏无明显毒性。Tunel检测的凋亡细胞呈棕色或棕黄色,白藜芦醇组和金纳米荷载白藜芦醇组凋亡细胞的比例分别为50.2%和65.8%,均比正常组明显增加。免疫组化检测结果也表明金纳米荷载白藜芦醇可显著抑制VEGF的表达。蛋白免疫印记法结果表明,Caspase家族相关凋亡蛋白和PI3K/Akt信号通路在促进肝癌细胞中发挥重要的作用,并且金纳米荷载白藜芦醇对肿瘤组织中caspase-8、caspase-9、caspase-3、bax、PI3K和Akt蛋白的调节作用强于白藜芦醇。体内外实验结果表明白藜芦醇和金纳米荷载白藜芦醇均可显著抑制Hepg2细胞和肝癌移植瘤体的生长,并且金纳米荷载白藜芦醇的抗肿瘤效果要优于白藜芦醇,其作用机制可能与调节线粒体相关凋亡蛋白有关。另外,金纳米荷载白藜芦醇对L02细胞、GMCs和裸鼠的脏器均没有明显的毒性作用。因此,金纳米荷载白藜芦醇作为治疗或者辅助治疗肝癌的药物可能会有潜在的价值。
Abstract
ai zheng zuo wei yi ge ri yi yan chong de quan qiu jian kang wen ti ,mei nian dou ying xiang zhao shu bai mo ren 。ji zhong ,gan ai shi shi jie shang di wu da chang jian e xing zhong liu ,ye shi dao zhi si wang de di er da yuan yin 。zai lin chuang yi sheng he ke xue jia de gong tong nu li xia ,gan ai de zong ti sheng cun lv de yi yan chang 。ran er ,zai zhi liao guo cheng zhong ,fang liao he hua liao yao wu zao cheng de de bu liang fan ying ,da da jiang di le huan zhe de sheng cun lv 。yin ci ,pai qie xu yao yan jiu xin de yao wu lai di gao wo men zhen duan 、yu fang he zhi liao gan ai de neng li 。na mi li zi zuo wei yun zai gong ju wei yi xue zhen duan he zhi liao di gong le ju da de qian li 。yin na mi he jin na mi ju you liang hao de sheng wu xiang rong xing 、di du xing he gong neng duo yang xing ,bing ju zai sheng wu yi xue ling yu you zhao an fan de ying yong 。you yu jin na mi ke li biao mian ji da ,neng bei duo chong zhi liao yao wu bao fu ,yin ci zai yao wu chuan di ji tong zhong ju you an kuo de ying yong qian jing 。zui jin de yan jiu biao ming ,jin na mi ke li yi bei bao dao ying yong yu kang zhong liu zhi liao ,bao gua ai zheng kang yuan he zuo ji de chuan di 。gen ju jin na mi ke li de te xing ,ben shi yan shi cai xiang na mi ji shu shi fou ke yi ying yong yu zeng jiang bai li lu chun de kang zhong liu zuo yong 。bai li lu chun shi yi chong tian ran de hua ge wu ,cun zai yu jiang guo 、jian guo 、hu zhang he pu tao zhong 。jin nian lai ,da liang de yan jiu biao ming bai li lu chun ju you duo chong yao li xue xiao ying ,bao gua qian zai de kang ai 、kang yan 、xin zang bao hu he shen jing bao hu huo xing 。dan shi ,bai li lu chun nan rong yu shui he sheng wu li yong du di de te dian da da xian zhi le ta de ying yong 。mu qian ,you hen duo yan jiu zhe tong guo dui bai li lu chun jin hang xiu shi lai gai shan ta de que dian 。ji yu zhe xie fa xian ,ben ke ti zai ti wai he ti nei jin hang le yi ji lie yan jiu ,yi tan tao jin na mi ke li shi fou ke yong yu zeng jiang Resde kang zhong liu zuo yong 。ben shi yan li yong bai li lu chun zi shen ju bei de hai yuan xing ,yi bai li lu chun he lv jin suan rong ye wei yuan liao ,zai 37℃、zhi liang bi wei 1:2、fan ying shi jian wei 120 minde tiao jian xia ,yi bu fa zhi bei li jing jun yi 、fen san xing liang hao de jin na mi he zai bai li lu chun 。zi wai guang pu jie guo biao ming jin na mi he zai bai li lu chun de deng li zi gong zhen xi shou feng zai 539 nmchu chu xian 。tou she dian jing shi yan jie guo biao ming tong guo diao jie fan ying wen du 、shi jian he zhi liang bi ke yi de dao li jing zai 39 nmde jin na mi he zai bai li lu chun 。ling wai ,gong wai guang pu shi yan shu ju biao ming jin yu bai li lu chun fen qiang ji jie gou zhong yang de xie tong zuo yong ,shi bai li lu chun bao guo zai jin na mi li zi biao mian ,bai li lu chun zhong de tan tan shuang jian he qiang ji de dao le bao liu ,bing ju yi you da liang yan jiu biao ming tan tan shuang jian he qiang ji zai bai li lu chun fa hui hen duo yao li zuo yong zhong qi dao chong yao de zuo yong 。yin ci ,wo men yu ce jin na mi he zai bai li lu chun ke neng ju you bai li lu chun de bu fen yao li zuo yong he jin na mi ke li de you liang te xing ,zai kang zhong liu fang mian ju you liang hao de ying yong qian jing 。MTTshi yan jian ce bai li lu chun he jin na mi he zai bai li lu chun dui Hepg2xi bao sheng chang de ying xiang ;AOran se fa jian ce yao wu dui gan ai xi bao xing tai de ying xiang ;liu shi xi bao yi jian ce gan ai xi bao de diao wang qing kuang ;tou she dian jing jian ce jin na mi he zai bai li lu chun zai Hepg2xi bao nei de fen bu qing kuang ;dan bai yin ji mian yi fa （Western Blot）jian ce Caspasejia zu he PI3K/Aktdeng xiang guan diao wang dan bai de bian hua ;MTTshi yan jian ce bai li lu chun he jin na mi he zai bai li lu chun dui zheng chang gan xi bao L02he shen xiao qiu ji mo GMCsxi bao de du xing zuo yong 。zai yi shang shi yan de ji chu shang ,yong BALB/cluo shu gou jian ren gan ai Hepg2xi bao he liu mo xing ,chi xu gei yu yao wu 42tian ,chu si he liu luo shu ,bao li liu ti ,HEran se guan cha zhong liu zu zhi de xing tai xue bian hua ;Tunelran se jian ce zhong liu zu zhi zhong xi bao diao wang de qing kuang ;mian yi zu hua fa jian ce liu ti zhong xie guan sheng chang yin zi （VEGF）de biao da 。zhai qu dong wu de xin zang 、gan zang 、pi zang 、fei zang he shen zang ,HEqie pian ran se guan cha shou shi wu dui shang shu qi guan zu zhi xing tai xue gai bian de ying xiang ,tong shi yong MTTfa jian ce yao wu dui L02he GMCsxi bao de du xing 。MTTjie guo biao ming bai li lu chun he jin na mi he zai bai li lu chun dui Hepg2xi bao jun ju you ming xian de yi zhi zuo yong ,bing ju jin na mi he zai bai li lu chun de yi zhi zuo yong geng xian zhe 。bai li lu chun he jin na mi he zai bai li lu chun chu li Hepg2xi bao 24 hhou ,ban shu yi zhi nong du (IC50)fen bie wei 27.74g/mlhe 3.84g/ml,wei le jin yi bu jian ce bai li lu chun he jin na mi he zai bai li lu chun dui L02xi bao he GMCsxi bao de du xing zuo yong ,fen bie yong bai li lu chun he jin na mi he zai bai li lu chun chu li 24 h,jin na mi he zai bai li lu chun dui L02xi bao he GMCsxi bao de yi zhi lv yi ci wei 35%he 10%,er bai li lu chun zu de yi zhi lv yi ci wei 83%he 63%。AOran se jie guo biao ming liang chong yao wu jun neng gou cu jin xi bao diao wang ,dao zhi xi bao fa sheng xing tai xue gai bian ,gei yao hou Hepg2xi bao de xi bao he mo zhou su ,xi bao ti ji bian xiao er cheng lu se de ying guang ,diao wang xi bao cheng sui pian zhuang 。sui zhao gei yao nong du de di zeng ,ji diao wang xi bao shu ye xiang ying zeng jia 。tou she dian jing jie guo biao ming gong neng hua de jin na mi he zai bai li lu chun zai xi bao nei da liang ji lei ,bing tong guo xian li ti tu jing you dao xi bao diao wang 。liu shi xi bao yi jie guo ye biao ming xi bao de diao wang you xian zhe de ji liang yi lai xing 。WBjian ce xiang guan dan bai fa xian ,jin na mi he zai bai li lu chun ke xian zhe shang diao ji huo de caspase-3、caspase-8、caspase-9yu baxhe xia diao PI3Kyu Aktde biao da 。bai li lu chun dui shang shu dan bai de diao jie ye cheng xiang tong qu shi ,dan diao jie zuo yong ruo yu jin na mi he zai bai li lu chun 。he liu shi yan jie shu hou ,bao li liu ti bing chen chong 。fa xian bai li lu chun he jin na mi he zai bai li lu chun dui liu ti de sheng chang jun you jiao jiang yi zhi zuo yong 。bai li lu chun zu he jin na mi he zai bai li lu chun zu zhong luo shu yi zhi liu ti de ti ji he chong liang jun ming xian de jiang di ,dui liu chong de yi zhi lv fen bie wei 35.5%he 57.2%。he liu zu zhi jing HEran se hou ,jie guo xian shi ,mo xing zu zhong liu zu zhi zhong de xi bao pai lie zheng ji ,xi bao xing tai wan zheng ;gei yao zu de xi bao sui zhao nong du de di zeng xi bao shu liang zhu jian jian shao ,xi bao he mo zhou su ,cheng jun yun lan se ,xiang jiao yu bai li lu chun zu ,jin na mi he zai bai li lu chun zu you dao xi bao diao wang de xiao guo geng jia xian zhe ;HEran se jie guo biao ming ,fa xian jin na mi he zai bai li lu chun dui luo shu de xin zang 、gan zang 、pi zang 、fei zang he shen zang mo ming xian du xing 。Tuneljian ce de diao wang xi bao cheng zong se huo zong huang se ,bai li lu chun zu he jin na mi he zai bai li lu chun zu diao wang xi bao de bi li fen bie wei 50.2%he 65.8%,jun bi zheng chang zu ming xian zeng jia 。mian yi zu hua jian ce jie guo ye biao ming jin na mi he zai bai li lu chun ke xian zhe yi zhi VEGFde biao da 。dan bai mian yi yin ji fa jie guo biao ming ,Caspasejia zu xiang guan diao wang dan bai he PI3K/Aktxin hao tong lu zai cu jin gan ai xi bao zhong fa hui chong yao de zuo yong ,bing ju jin na mi he zai bai li lu chun dui zhong liu zu zhi zhong caspase-8、caspase-9、caspase-3、bax、PI3Khe Aktdan bai de diao jie zuo yong jiang yu bai li lu chun 。ti nei wai shi yan jie guo biao ming bai li lu chun he jin na mi he zai bai li lu chun jun ke xian zhe yi zhi Hepg2xi bao he gan ai yi zhi liu ti de sheng chang ,bing ju jin na mi he zai bai li lu chun de kang zhong liu xiao guo yao you yu bai li lu chun ,ji zuo yong ji zhi ke neng yu diao jie xian li ti xiang guan diao wang dan bai you guan 。ling wai ,jin na mi he zai bai li lu chun dui L02xi bao 、GMCshe luo shu de zang qi jun mei you ming xian de du xing zuo yong 。yin ci ,jin na mi he zai bai li lu chun zuo wei zhi liao huo zhe fu zhu zhi liao gan ai de yao wu ke neng hui you qian zai de jia zhi 。
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论文作者分别是来自西南大学的张朵朵，发表于刊物西南大学2019-09-24论文，是一篇关于白藜芦醇论文,金纳米荷载白藜芦醇论文,表征论文,细胞系论文,移植瘤论文,西南大学2019-09-24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，资料来自西南大学2019-09-24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。
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