缓释纳米尿激酶的制备VIP专享VIP免费

上海交通大学学报(医学版)JournalofShanghaiJiaotongUniversity(MedicalScience)Vol．31No．6Jun．2011［基金项目］上海市科委课题(0852nm05100)(ShanghaiScienceandTechnologyCommitteeFoundation，0852nm05100)。［作者简介］金海将(1984—)，男，博士生;电子信箱:jinhaijiang521@163．com。［通信作者］张皓，电子信箱:chlzhcx@163．com。［文章编号］1674-8115(2011)06-0862-04·技术与方法·缓释纳米尿激酶的制备金海将，张皓，张柏根，孙敏莉(上海交通大学医学院附属仁济医院血管外科，上海200001)［摘要］目的制备包载尿激酶的水溶性壳聚糖(WSC)纳米粒子，观察纳米尿激酶的活性。方法采用离子交联法制备包载尿激酶的WSC纳米粒子;检测尿激酶纳米粒子的包封率，分析WSC与三聚磷酸钠质量比、尿激酶浓度、磁力搅拌速度和时间、超声时间和功率等条件对包封率的影响，获得包封率最高的制备条件;粒径仪测量尿激酶纳米粒子粒径;超滤离心法纯化尿激酶纳米粒子，注入新西兰兔体内(纳米尿激酶组)，于不同时间点取血测定尿激酶活性，以注射单纯尿激酶的动物作为对照组。结果室温下，将三聚磷酸钠溶液(0．6g/L)滴加入包含尿激酶(1mg/10mL)的WSC溶液(1．0g/L)，磁力搅拌速度800r/min，滴加完毕后继续搅拌60min，冰浴条件下超声30s(功率10W)，成功制备出包封率为94．8%的尿激酶纳米粒子;对照组血尿激酶活性达到峰值后随着时间的延长逐渐降低，纳米尿激酶组血尿激酶活性达到峰值后仍保持较高水平。结论成功制备了包载尿激酶的WSC纳米粒子，纳米尿激酶具有缓释功能并保持了尿激酶的活性。［关键词］水溶性壳聚糖;尿激酶纳米粒子;包封率;离子交联法［DOI］10．3969/j．issn．1674-8115．2011．06．041［中图分类号］TB383［文献标志码］BPreparationofsustained-releaseurokinasenanoparticlesJINHai-jiang，ZHANGHao，ZHANGBai-gen，SUNMin-li(DepartmentofVascularSurgery，RenjiHospital，ShanghaiJiaotongUniversitySchoolofMedicine，Shanghai200001，China)［Abstract］ObjectiveToprepareurokinasewatersolublechitosan(WSC)nanoparticles，andinvestigateitsactivity．MethodsUrokinaseWSCnanoparticleswerepreparedbyioniccrosslinkingmethod，andtheencapsulationefficiencyofurokinasenanoparticleswasmeasured．TheeffectsofqualityratioofWSCtotripolyphosphate，concentrationofurokinase，timeandvelocityofmagneticstirring，andtimeandpowerofultrasoundonencapsulationefficiencyweredetermined，andtheconditionsforthehighestencapsulationefficiencywereobtained．Thediametersofurokinasenanoparticlesweremeasuredbyparticlesizeinstrument．Urokinasenanoparticleswerepurifiedbycentrifugalultrafiltration，andwereinjectedintoNewZealandrabbits(urokinasenanoparticlesgroup)．Bloodsamplesweretakenfromrabbitsatdifferenttimepoints，andtheactivityofurokinasenanoparticleswasexamined．Rabbitsinjectedwithsingleurokinasewereservedascontrolgroup．ResultsUnderroomtemperature，whentripolyphosphatesolution(0．6g/L)wasdroppedintoWSCsolution(1．0g/L)containingurokinase(1mg/10mL)，urokinasenanoparticleswiththehighestencapsulationefficiency(94．8%)wereobtainedwiththetimeandvelocityofstirringof60minand800r/minrespectively，andthetimeandpowerofultrasoundinicebathof30sand10Wrespectively．Theserumactivityofurokinasegraduallydecreasedwithtimeafterreachingthepeakincontrolgroup，whilethatmaintainedatahigherlevelafterreachingthepeakinurokinasenanoparticlesgroup．ConclusionUrokinaseWSCnanoparticleshavebeensuccessfullyprepared，whichpossessnotonlysustained-releasefunction，butalsoactivityofurokinase．［Keywords］watersolublechitosan;urokinasenanoparticles;encapsulationefficiency;ioniccrosslinkingmethod临床上常用尿激酶(urokinase)治疗血栓性疾病，但尿激酶半衰期短，需持续大量给药，故实际应用不便...