本文主要研究内容
作者徐晓玲（2019）在《基于唾液酸的类风湿关节炎靶向给药系统研究》一文中研究指出：类风湿关节炎(Rheumatoid Arthritis,RA)是一种以多关节滑膜炎症为主要特征的系统性自身免疫疾病,常伴关节外器官受累及血清类风湿因子阳性,最终导致关节畸形及功能丧失,高发于30-50岁的女性,发病率呈逐年上升的趋势。目前临床上RA的治疗以药物治疗和手术治疗为主,但手术治疗易造成关节腔二次损伤,滑膜的切除使关节失去了合成滑液的基本功能,进一步加剧了关节功能的受损。甲氨蝶呤(Methotrexate,MTX)是临床一线的抗风湿药,具有免疫抑制、抗炎作用,能够减轻疼痛、缓解症状、控制病情发展。但因其在体内的非特异性分布,使得进入炎症关节部位的药物分子数量有限,而在正常组织中大量分布,服用高剂量甲氨蝶呤常产生溃疡性口腔炎、腹泻甚至致命的肠穿孔等毒副作用,严重影响了患者的生活质量,而低剂量的治疗又严重制约RA疗效,且长期小剂量使用仅能延缓骨质侵蚀的进展,而不能阻止长期炎症反应导致的骨质疏松的发生,更不能促进已受损骨质的修复与再生。新型纳米给药系统是改善药物体内分布,降低药物毒副作用的有效手段,且可通过载体材料的功能化实现RA的多靶点治疗。唾液酸(SialicAcid,SA)是一类位于细胞膜最外层的九碳糖,可与E选择素特异性结合,且唾液酸化的糖复合物可通过抑制破骨细胞成熟,促进成骨细胞分化,进而促进骨的修复和再生。本研究采用唾液酸、葡聚糖和硬脂酸为原料合成了唾液酸-葡聚糖-硬脂酸(SA-Dex-OA,SDO)嫁接物;SDO可在水性介质中自组装形成胶束,临界胶束浓度(Critical micelle concentration,CMC)为 55.06 μg/mL,胶束粒径为75.20 ± 11.91 nm,胶束能有效负载抗类风湿药物MTX,包封率为89.39 ±0.26%。唾液酸-葡聚糖-硬脂酸载药胶束(SA-Dex-OA/MTX,SDM)粒径为117.33±5.77 nm,载药胶束对药物有明显的缓释效应,药物可持续释放48 h以上。以人脐静脉内皮细胞(HUVEC)为模型细胞,对嫁接物的细胞毒性进行考察,结果显示其在1-1000 μg/mL浓度范围内对细胞的生长及增殖均无影响,细胞毒性较低。采用脂多糖刺激HUVEC构建E-选择素受体高表达的体外炎症细胞模型用于考察SDO嫁接物胶束的主动靶向能力,以葡聚糖-硬脂酸(Dex-OA,DO)嫁接物胶束为对照,结果表明炎症HUVEC对SDO嫁接物胶束具有更好的摄取行为;采用游离唾液酸阻断细胞表面E-选择素受体后,炎症细胞对SDO嫁接物胶束的摄取量降低;E-选择素荧光与SDO胶束荧光信号呈现共定位现象,证实SDO嫁接物胶束的炎症血管内皮细胞主动转运是由SA与E-选择素的特异性结合所介导的。以小鼠胚胎成骨细胞前体细胞(MC3T3-E1)为模型细胞,对嫁接物的骨修复与再生作用进行考察,结果显示SDO嫁接物胶束可通过唾液酸介导的促胞内碱性磷酸酶活力和胞外矿化结节形成,促进成骨细胞分化和矿化。采用胶原诱导关节炎法构建大鼠RA模型,尾静脉给药考察包载荧光探针的嫁接物胶束在模型动物体内的分布情况,结果显示SDO嫁接物胶束可通过唾液酸与炎症关节周围血管内皮细胞表面高表达E选择素的特异性结合,实现在炎症关节部位的大量蓄积。经尾静脉注射给药14天后,SDM组足爪肿胀程度显著减轻,关节炎评分指数显著下降;滑膜组织增生和胞内线粒体肿胀情况得到有效抑制,显著改善了细胞缺血缺氧状态;同时炎性细胞浸润程度明显减轻,TNF-a和IL-6的合成和释放能力显著降低,炎症反应得到有效抑制。另外,SDM载药胶束还可通过唾液酸介导的骨修复调控作用,上调OPG表达,下调RANKL表达,使骨代谢向骨形成倾斜,提高骨密度,发挥骨修复与再生作用;MTX给药14天后,血清中谷草转氨酶、谷丙转氨酶、尿素氮和血肌酐的浓度显著升高,对肝肾均有一定毒性,而SDM组生化指标与正常组接近,显示SDM载药胶束的安全性。纳米羟基磷灰石(Hydroxyapatite,HAP)作为骨骼的天然组分之一,具有生物粘附性强、可吸附药量大、酸敏感、骨诱导性能良好等特性,是优良的人体骨仿生再生材料。本研究在前期研究的基础上,以具有修复软骨作用的壳寡糖为胶束主要材料,通过酰胺反应合成唾液酸-壳寡糖-脱氧胆酸嫁接物胶束(SA-CSO-DA,SD);SD可在水性介质中自组装形成胶束,CMC为38.45 μg/mL;以具有较好表面活性剂特性的该嫁接物胶束作为稳定剂,通过水热法制得棒状的唾液酸修饰壳寡糖/羟基磷灰石(SH)纳米粒,大小均一,粒径约为30.03±6.54nm;该纳米粒可有效吸附MTX,制得的载药纳米粒(SHM)外观仍呈棒状,分布均一,粒径大小增加至46.08 ± 7.88 nm,包封率为68.67±2.80%。体外释放结果显示,SHM呈现明显的酸敏感释放特性,在pH 7.4中性介质中释放缓慢,而在pH 5.5酸性条件下释放加快,48h内的累积释放百分率可达87.95%左右。以原代大鼠软骨细胞为模型细胞,考察SH纳米粒的促软骨修复作用,结果表明,SH纳米粒具有较低的细胞毒性;IL-1β刺激的原代大鼠软骨细胞经SH处理后,凋亡细胞的比例显著降低,糖胺多糖含量显著增加,Ⅱ型胶原表达明显上调,而HAP预处理基本不影响凋亡细胞比例、糖胺多糖含量和Ⅱ型胶原的表达。以原代成骨细胞为模型细胞,考察SH纳米粒的促成骨修复与再生作用,结果显示,与SH共孵育后,胞内碱性磷酸酶活力明显增强,矿化结节数目显著增加,Ⅰ型胶原和OPG表达明显上调,显著促进了成骨细胞的分化及矿化进程。采用胶原诱导关节炎法建立RA模型,SH纳米粒经静脉给药后,其可有效分布至靶部位,实现在炎症关节部位的大量蓄积;经尾静脉注射给药28天后,SHM组足爪肿胀程度显著减轻,关节炎评分指数显著下降;脾脏指数和胸腺指数有效降低,免疫功能亢进情况得到显著改善,同时滑膜组织中炎性细胞合成和释放TNF-α和IL-6的量显著降低,炎症反应得到有效抑制;还可通过补充软骨组织中糖胺多糖含量、上调Ⅱ型胶原表达及抑制基质金属蛋白酶的活性,发挥促软骨修复作用;SHM纳米给药系统可通过上调骨形成阶段关键蛋白OPG、下调破骨细胞激活因子RANKL,使新骨生成速度显著加快,显著增加骨质及骨量,实现骨修复与再生作用;MTX给药28天后,血清中尿素氮的浓度和谷草转氨酶/谷丙转氨酶比值显著升高,对肝肾均有一定毒性,而SHM组生化指标与正常组接近,肝脏和肾脏无明显病变区域,显示SHM纳米给药系统安全性较高。本研究构建的功能性甲氨蝶呤纳米给药系统均能通过药物向炎症关节部位的靶向递送,实现类风湿关节炎炎症反应的有效抑制,同时,残留的壳寡糖可促进软骨修复;纳米羟基磷灰石协同唾液酸,发挥成骨修复与再生功能,最终实现类风湿性关节炎的安全高效治疗,为RA治疗方法提供新策略和新思路。
Abstract
lei feng shi guan jie yan (Rheumatoid Arthritis,RA)shi yi chong yi duo guan jie hua mo yan zheng wei zhu yao te zheng de ji tong xing zi shen mian yi ji bing ,chang ban guan jie wai qi guan shou lei ji xie qing lei feng shi yin zi yang xing ,zui zhong dao zhi guan jie ji xing ji gong neng sang shi ,gao fa yu 30-50sui de nv xing ,fa bing lv cheng zhu nian shang sheng de qu shi 。mu qian lin chuang shang RAde zhi liao yi yao wu zhi liao he shou shu zhi liao wei zhu ,dan shou shu zhi liao yi zao cheng guan jie qiang er ci sun shang ,hua mo de qie chu shi guan jie shi qu le ge cheng hua ye de ji ben gong neng ,jin yi bu jia ju le guan jie gong neng de shou sun 。jia an die ling (Methotrexate,MTX)shi lin chuang yi xian de kang feng shi yao ,ju you mian yi yi zhi 、kang yan zuo yong ,neng gou jian qing teng tong 、huan jie zheng zhuang 、kong zhi bing qing fa zhan 。dan yin ji zai ti nei de fei te yi xing fen bu ,shi de jin ru yan zheng guan jie bu wei de yao wu fen zi shu liang you xian ,er zai zheng chang zu zhi zhong da liang fen bu ,fu yong gao ji liang jia an die ling chang chan sheng hui yang xing kou qiang yan 、fu xie shen zhi zhi ming de chang chuan kong deng du fu zuo yong ,yan chong ying xiang le huan zhe de sheng huo zhi liang ,er di ji liang de zhi liao you yan chong zhi yao RAliao xiao ,ju chang ji xiao ji liang shi yong jin neng yan huan gu zhi qin shi de jin zhan ,er bu neng zu zhi chang ji yan zheng fan ying dao zhi de gu zhi shu song de fa sheng ,geng bu neng cu jin yi shou sun gu zhi de xiu fu yu zai sheng 。xin xing na mi gei yao ji tong shi gai shan yao wu ti nei fen bu ,jiang di yao wu du fu zuo yong de you xiao shou duan ,ju ke tong guo zai ti cai liao de gong neng hua shi xian RAde duo ba dian zhi liao 。tuo ye suan (SialicAcid,SA)shi yi lei wei yu xi bao mo zui wai ceng de jiu tan tang ,ke yu Eshua ze su te yi xing jie ge ,ju tuo ye suan hua de tang fu ge wu ke tong guo yi zhi po gu xi bao cheng shou ,cu jin cheng gu xi bao fen hua ,jin er cu jin gu de xiu fu he zai sheng 。ben yan jiu cai yong tuo ye suan 、pu ju tang he ying zhi suan wei yuan liao ge cheng le tuo ye suan -pu ju tang -ying zhi suan (SA-Dex-OA,SDO)jia jie wu ;SDOke zai shui xing jie zhi zhong zi zu zhuang xing cheng jiao shu ,lin jie jiao shu nong du (Critical micelle concentration,CMC)wei 55.06 μg/mL,jiao shu li jing wei 75.20 ± 11.91 nm,jiao shu neng you xiao fu zai kang lei feng shi yao wu MTX,bao feng lv wei 89.39 ±0.26%。tuo ye suan -pu ju tang -ying zhi suan zai yao jiao shu (SA-Dex-OA/MTX,SDM)li jing wei 117.33±5.77 nm,zai yao jiao shu dui yao wu you ming xian de huan shi xiao ying ,yao wu ke chi xu shi fang 48 hyi shang 。yi ren qi jing mai nei pi xi bao (HUVEC)wei mo xing xi bao ,dui jia jie wu de xi bao du xing jin hang kao cha ,jie guo xian shi ji zai 1-1000 μg/mLnong du fan wei nei dui xi bao de sheng chang ji zeng shi jun mo ying xiang ,xi bao du xing jiao di 。cai yong zhi duo tang ci ji HUVECgou jian E-shua ze su shou ti gao biao da de ti wai yan zheng xi bao mo xing yong yu kao cha SDOjia jie wu jiao shu de zhu dong ba xiang neng li ,yi pu ju tang -ying zhi suan (Dex-OA,DO)jia jie wu jiao shu wei dui zhao ,jie guo biao ming yan zheng HUVECdui SDOjia jie wu jiao shu ju you geng hao de she qu hang wei ;cai yong you li tuo ye suan zu duan xi bao biao mian E-shua ze su shou ti hou ,yan zheng xi bao dui SDOjia jie wu jiao shu de she qu liang jiang di ;E-shua ze su ying guang yu SDOjiao shu ying guang xin hao cheng xian gong ding wei xian xiang ,zheng shi SDOjia jie wu jiao shu de yan zheng xie guan nei pi xi bao zhu dong zhuai yun shi you SAyu E-shua ze su de te yi xing jie ge suo jie dao de 。yi xiao shu pei tai cheng gu xi bao qian ti xi bao (MC3T3-E1)wei mo xing xi bao ,dui jia jie wu de gu xiu fu yu zai sheng zuo yong jin hang kao cha ,jie guo xian shi SDOjia jie wu jiao shu ke tong guo tuo ye suan jie dao de cu bao nei jian xing lin suan mei huo li he bao wai kuang hua jie jie xing cheng ,cu jin cheng gu xi bao fen hua he kuang hua 。cai yong jiao yuan you dao guan jie yan fa gou jian da shu RAmo xing ,wei jing mai gei yao kao cha bao zai ying guang tan zhen de jia jie wu jiao shu zai mo xing dong wu ti nei de fen bu qing kuang ,jie guo xian shi SDOjia jie wu jiao shu ke tong guo tuo ye suan yu yan zheng guan jie zhou wei xie guan nei pi xi bao biao mian gao biao da Eshua ze su de te yi xing jie ge ,shi xian zai yan zheng guan jie bu wei de da liang xu ji 。jing wei jing mai zhu she gei yao 14tian hou ,SDMzu zu zhao zhong zhang cheng du xian zhe jian qing ,guan jie yan ping fen zhi shu xian zhe xia jiang ;hua mo zu zhi zeng sheng he bao nei xian li ti zhong zhang qing kuang de dao you xiao yi zhi ,xian zhe gai shan le xi bao que xie que yang zhuang tai ;tong shi yan xing xi bao jin run cheng du ming xian jian qing ,TNF-ahe IL-6de ge cheng he shi fang neng li xian zhe jiang di ,yan zheng fan ying de dao you xiao yi zhi 。ling wai ,SDMzai yao jiao shu hai ke tong guo tuo ye suan jie dao de gu xiu fu diao kong zuo yong ,shang diao OPGbiao da ,xia diao RANKLbiao da ,shi gu dai xie xiang gu xing cheng qing xie ,di gao gu mi du ,fa hui gu xiu fu yu zai sheng zuo yong ;MTXgei yao 14tian hou ,xie qing zhong gu cao zhuai an mei 、gu bing zhuai an mei 、niao su dan he xie ji gan de nong du xian zhe sheng gao ,dui gan shen jun you yi ding du xing ,er SDMzu sheng hua zhi biao yu zheng chang zu jie jin ,xian shi SDMzai yao jiao shu de an quan xing 。na mi qiang ji lin hui dan (Hydroxyapatite,HAP)zuo wei gu ge de tian ran zu fen zhi yi ,ju you sheng wu nian fu xing jiang 、ke xi fu yao liang da 、suan min gan 、gu you dao xing neng liang hao deng te xing ,shi you liang de ren ti gu fang sheng zai sheng cai liao 。ben yan jiu zai qian ji yan jiu de ji chu shang ,yi ju you xiu fu ruan gu zuo yong de ke gua tang wei jiao shu zhu yao cai liao ,tong guo xian an fan ying ge cheng tuo ye suan -ke gua tang -tuo yang dan suan jia jie wu jiao shu (SA-CSO-DA,SD);SDke zai shui xing jie zhi zhong zi zu zhuang xing cheng jiao shu ,CMCwei 38.45 μg/mL;yi ju you jiao hao biao mian huo xing ji te xing de gai jia jie wu jiao shu zuo wei wen ding ji ,tong guo shui re fa zhi de bang zhuang de tuo ye suan xiu shi ke gua tang /qiang ji lin hui dan (SH)na mi li ,da xiao jun yi ,li jing yao wei 30.03±6.54nm;gai na mi li ke you xiao xi fu MTX,zhi de de zai yao na mi li (SHM)wai guan reng cheng bang zhuang ,fen bu jun yi ,li jing da xiao zeng jia zhi 46.08 ± 7.88 nm,bao feng lv wei 68.67±2.80%。ti wai shi fang jie guo xian shi ,SHMcheng xian ming xian de suan min gan shi fang te xing ,zai pH 7.4zhong xing jie zhi zhong shi fang huan man ,er zai pH 5.5suan xing tiao jian xia shi fang jia kuai ,48hnei de lei ji shi fang bai fen lv ke da 87.95%zuo you 。yi yuan dai da shu ruan gu xi bao wei mo xing xi bao ,kao cha SHna mi li de cu ruan gu xiu fu zuo yong ,jie guo biao ming ,SHna mi li ju you jiao di de xi bao du xing ;IL-1βci ji de yuan dai da shu ruan gu xi bao jing SHchu li hou ,diao wang xi bao de bi li xian zhe jiang di ,tang an duo tang han liang xian zhe zeng jia ,Ⅱxing jiao yuan biao da ming xian shang diao ,er HAPyu chu li ji ben bu ying xiang diao wang xi bao bi li 、tang an duo tang han liang he Ⅱxing jiao yuan de biao da 。yi yuan dai cheng gu xi bao wei mo xing xi bao ,kao cha SHna mi li de cu cheng gu xiu fu yu zai sheng zuo yong ,jie guo xian shi ,yu SHgong fu yo hou ,bao nei jian xing lin suan mei huo li ming xian zeng jiang ,kuang hua jie jie shu mu xian zhe zeng jia ,Ⅰxing jiao yuan he OPGbiao da ming xian shang diao ,xian zhe cu jin le cheng gu xi bao de fen hua ji kuang hua jin cheng 。cai yong jiao yuan you dao guan jie yan fa jian li RAmo xing ,SHna mi li jing jing mai gei yao hou ,ji ke you xiao fen bu zhi ba bu wei ,shi xian zai yan zheng guan jie bu wei de da liang xu ji ;jing wei jing mai zhu she gei yao 28tian hou ,SHMzu zu zhao zhong zhang cheng du xian zhe jian qing ,guan jie yan ping fen zhi shu xian zhe xia jiang ;pi zang zhi shu he xiong xian zhi shu you xiao jiang di ,mian yi gong neng kang jin qing kuang de dao xian zhe gai shan ,tong shi hua mo zu zhi zhong yan xing xi bao ge cheng he shi fang TNF-αhe IL-6de liang xian zhe jiang di ,yan zheng fan ying de dao you xiao yi zhi ;hai ke tong guo bu chong ruan gu zu zhi zhong tang an duo tang han liang 、shang diao Ⅱxing jiao yuan biao da ji yi zhi ji zhi jin shu dan bai mei de huo xing ,fa hui cu ruan gu xiu fu zuo yong ;SHMna mi gei yao ji tong ke tong guo shang diao gu xing cheng jie duan guan jian dan bai OPG、xia diao po gu xi bao ji huo yin zi RANKL,shi xin gu sheng cheng su du xian zhe jia kuai ,xian zhe zeng jia gu zhi ji gu liang ,shi xian gu xiu fu yu zai sheng zuo yong ;MTXgei yao 28tian hou ,xie qing zhong niao su dan de nong du he gu cao zhuai an mei /gu bing zhuai an mei bi zhi xian zhe sheng gao ,dui gan shen jun you yi ding du xing ,er SHMzu sheng hua zhi biao yu zheng chang zu jie jin ,gan zang he shen zang mo ming xian bing bian ou yu ,xian shi SHMna mi gei yao ji tong an quan xing jiao gao 。ben yan jiu gou jian de gong neng xing jia an die ling na mi gei yao ji tong jun neng tong guo yao wu xiang yan zheng guan jie bu wei de ba xiang di song ,shi xian lei feng shi guan jie yan yan zheng fan ying de you xiao yi zhi ,tong shi ,can liu de ke gua tang ke cu jin ruan gu xiu fu ;na mi qiang ji lin hui dan xie tong tuo ye suan ,fa hui cheng gu xiu fu yu zai sheng gong neng ,zui zhong shi xian lei feng shi xing guan jie yan de an quan gao xiao zhi liao ,wei RAzhi liao fang fa di gong xin ce lve he xin sai lu 。
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论文作者分别是来自浙江大学的徐晓玲，发表于刊物浙江大学2019-05-09论文，是一篇关于类风湿关节炎论文,唾液酸论文,羟基磷灰石论文,壳寡糖论文,靶向给药论文,协同治疗论文,骨再生论文,浙江大学2019-05-09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，资料来自浙江大学2019-05-09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。
标签：类风湿关节炎论文; 唾液酸论文; 羟基磷灰石论文; 壳寡糖论文; 靶向给药论文; 协同治疗论文; 骨再生论文; 浙江大学2019-05-09论文;