中华眼科杂志
中華眼科雜誌
중화안과잡지
Chinese Journal of Ophthalmology
2009年
11期
1027-1032
,共6页
詹文芳%蔡善君%刘锐%谢兵%李红%宿罡
詹文芳%蔡善君%劉銳%謝兵%李紅%宿罡
첨문방%채선군%류예%사병%리홍%숙강
胶囊%壳聚糖%内皮抑素类%投药,局部
膠囊%殼聚糖%內皮抑素類%投藥,跼部
효낭%각취당%내피억소류%투약,국부
Capsules%Chitosan%Endostatins%Administration,topical
目的 检测乳化.内部凝胶化制得的海藻酸钙.壳聚糖(CAC)内皮抑素微囊的体外释放情况,大鼠眼球周注射后眼内组织分布情况及生物相容性.方法 实验研究.采用乳化.内部凝胶化方式制备微囊,紫外分光光度法、考马斯亮蓝法检测内皮抑素囊外释放情况;32只SD大鼠随机分成4组,每组8只.A组:微囊化Es组,球周注射微囊化内皮抑素20 μl(2.5 g/L);B组:单纯内皮抑素组,球周注射内皮抑素蛋白20 μl(2.5 g/L);C组:空微囊组,球周注射空微囊20 μl;D组:生理盐水组,球周注射生理盐水20 μl.免疫组织化学法、ELISA法、Western-blot法检测内皮抑素分布、浓度和蛋白表达;观察SD大鼠的日常活动,进食情况和体重,以断颈法将大鼠处死取心、肝、脾、肺、肾及球周组织(球周注射部位周围组织)行病理学检查.采用独立样本设计定量资料t检验对数据进行分析.结果 十二烷基硫酸钠-聚丙烯酰胺凝胶电泳检测可见内皮抑素蛋白条带,3、7、14、21 d各时点上清液中内皮抑素蛋白质浓度逐渐增加.A组:球周注射后7、14 d,内外核层和RPE层见内皮抑素蛋白阳性表达;B组:7 d时,以上组织可见内皮抑素蛋白阳性表达,14 d后无蛋白阳性表达;C组和D组7、14 d以上:组织均未见内皮抑素蛋白阳性表达,表明内皮抑素蛋白经CAC微囊化后通过球周注射可以穿过巩膜壁进入球内.A组7、14 d,眼球组织匀浆内皮抑素蛋白浓度分别为每只眼(63.16±7.64)μl和(33.2±5.77)μg/L,而B组分别为(33.2±2.89) μg/L,(15.73±2.08)μg/L,A组明显大于B组且差异具有统计学意义(t=6.364、4.920,P=0.003、0.008);大鼠日常活动和进食情况均正常,14 d后病理学检查心肝、脾、肺、肾及球周组织未见异常.结论 微囊化内皮抑素具有缓控释功能,球周注射微囊化内皮抑素,能穿过巩膜壁并分布在视网膜组织内,微囊对机体无明显毒副作用.
目的 檢測乳化.內部凝膠化製得的海藻痠鈣.殼聚糖(CAC)內皮抑素微囊的體外釋放情況,大鼠眼毬週註射後眼內組織分佈情況及生物相容性.方法 實驗研究.採用乳化.內部凝膠化方式製備微囊,紫外分光光度法、攷馬斯亮藍法檢測內皮抑素囊外釋放情況;32隻SD大鼠隨機分成4組,每組8隻.A組:微囊化Es組,毬週註射微囊化內皮抑素20 μl(2.5 g/L);B組:單純內皮抑素組,毬週註射內皮抑素蛋白20 μl(2.5 g/L);C組:空微囊組,毬週註射空微囊20 μl;D組:生理鹽水組,毬週註射生理鹽水20 μl.免疫組織化學法、ELISA法、Western-blot法檢測內皮抑素分佈、濃度和蛋白錶達;觀察SD大鼠的日常活動,進食情況和體重,以斷頸法將大鼠處死取心、肝、脾、肺、腎及毬週組織(毬週註射部位週圍組織)行病理學檢查.採用獨立樣本設計定量資料t檢驗對數據進行分析.結果 十二烷基硫痠鈉-聚丙烯酰胺凝膠電泳檢測可見內皮抑素蛋白條帶,3、7、14、21 d各時點上清液中內皮抑素蛋白質濃度逐漸增加.A組:毬週註射後7、14 d,內外覈層和RPE層見內皮抑素蛋白暘性錶達;B組:7 d時,以上組織可見內皮抑素蛋白暘性錶達,14 d後無蛋白暘性錶達;C組和D組7、14 d以上:組織均未見內皮抑素蛋白暘性錶達,錶明內皮抑素蛋白經CAC微囊化後通過毬週註射可以穿過鞏膜壁進入毬內.A組7、14 d,眼毬組織勻漿內皮抑素蛋白濃度分彆為每隻眼(63.16±7.64)μl和(33.2±5.77)μg/L,而B組分彆為(33.2±2.89) μg/L,(15.73±2.08)μg/L,A組明顯大于B組且差異具有統計學意義(t=6.364、4.920,P=0.003、0.008);大鼠日常活動和進食情況均正常,14 d後病理學檢查心肝、脾、肺、腎及毬週組織未見異常.結論 微囊化內皮抑素具有緩控釋功能,毬週註射微囊化內皮抑素,能穿過鞏膜壁併分佈在視網膜組織內,微囊對機體無明顯毒副作用.
목적 검측유화.내부응효화제득적해조산개.각취당(CAC)내피억소미낭적체외석방정황,대서안구주주사후안내조직분포정황급생물상용성.방법 실험연구.채용유화.내부응효화방식제비미낭,자외분광광도법、고마사량람법검측내피억소낭외석방정황;32지SD대서수궤분성4조,매조8지.A조:미낭화Es조,구주주사미낭화내피억소20 μl(2.5 g/L);B조:단순내피억소조,구주주사내피억소단백20 μl(2.5 g/L);C조:공미낭조,구주주사공미낭20 μl;D조:생리염수조,구주주사생리염수20 μl.면역조직화학법、ELISA법、Western-blot법검측내피억소분포、농도화단백표체;관찰SD대서적일상활동,진식정황화체중,이단경법장대서처사취심、간、비、폐、신급구주조직(구주주사부위주위조직)행병이학검사.채용독립양본설계정량자료t검험대수거진행분석.결과 십이완기류산납-취병희선알응효전영검측가견내피억소단백조대,3、7、14、21 d각시점상청액중내피억소단백질농도축점증가.A조:구주주사후7、14 d,내외핵층화RPE층견내피억소단백양성표체;B조:7 d시,이상조직가견내피억소단백양성표체,14 d후무단백양성표체;C조화D조7、14 d이상:조직균미견내피억소단백양성표체,표명내피억소단백경CAC미낭화후통과구주주사가이천과공막벽진입구내.A조7、14 d,안구조직균장내피억소단백농도분별위매지안(63.16±7.64)μl화(33.2±5.77)μg/L,이B조분별위(33.2±2.89) μg/L,(15.73±2.08)μg/L,A조명현대우B조차차이구유통계학의의(t=6.364、4.920,P=0.003、0.008);대서일상활동화진식정황균정상,14 d후병이학검사심간、비、폐、신급구주조직미견이상.결론 미낭화내피억소구유완공석공능,구주주사미낭화내피억소,능천과공막벽병분포재시망막조직내,미낭대궤체무명현독부작용.
Objective To detect the release of endostatin in microcapsules of calcium alginate gel by emulsification-internal gelatification technology, to observe the distribution of Endostatin microcapsulized in the eye by periocular injection and biocompatibility.Methods The calcium alginate gel microcapsules encapsulating endostatin were prepared by emulsification-internal gelatification technology, the release of endostatin in microcapsules in vitro was examined by uv-Spectrophotometry.32 SD mice were divided into randomly four groups: group A, periocular injection of endostatin microcapsulized 20 μl (2.4 g/L) ; group B, periocular injection of endostatin protein 20 μl (2.5 g/L) ; group C, periocular injection of null-Microcapsules 20 μl; group D, periocular injection of saline 20 μl.Western blot, immunohistochemical and enzyme-linked immunosorbent assay were used.Protein expression and pathology of the heart, liver, spleen,lung, kidney and periocular tissue were examed.Daily activities and eating condition were observed, t-test was wsed to analyze the data.Results SDS polyacrylamidedel electrophoresis showed strap of ES protein.Concentration of ES protein in superior schedule fluid gradually increased at 3,7,14,21 days.A group: ES protein expression were observed in inner nuclear layer,outer nuclear layer and RPE lamina after periocular injection 7 or 14 days.B group: ES protein expression were observed in above tissues at 7 days, but not at 14 days.C and D group: ES protein expression were not observed in above tissues at 7 or 14 days.The results showed that Endostatin microcapsulized was able to pass through the sclera and spread out in various retinal tissues.Concertration of endostatin in the homogenates of eyeball was (63.16±7.64) μg · L~(-1)·eye~(-1) and (33.2±5.77) μg · L~(-1) · eye~(-1) respectively after one and two weeks in A group, but(33.2±2.89) μg · L~(-1) · eye~(-1) and (15.73±2.08) μg · L~(-1) · eye~(-1).The concertration of endostatin was significant difference in two group, A group was obviously larger than B group (t=6.364 and 4.920, P=0.003 and 0.008 respectively).Eriocular administration.Daily activities and eating condition were normal.The abnormality in important organs were not detected by pathologic examination.Conclusions Endostatin microcapsulized is able to release persistently, to pass through the sclera and spread out in various retinal tissues.There were good biocompatibility and not ill effect remarkably.
