本文關(guān)鍵詞： 細(xì)胞穿膜肽 聚精氨酸 眼球屏障 晶狀體上皮細(xì)胞　出處：《眼科新進(jìn)展》2017年06期 　論文類型：期刊論文

【摘要】：目的探討以多聚精氨酸為載體,是否能夠?qū)⑼庠吹鞍踪|(zhì)穿透細(xì)胞膜甚至穿透眼球屏障。方法本研究以9聚精氨酸(R9)為穿膜肽,構(gòu)建R9-綠色熒光蛋白(green fluorescent protein,GFP)、GFP兩種重組蛋白。體外實(shí)驗(yàn)中,將GFP、R9-GFP與人晶狀體上皮細(xì)胞(human lens epithelial cell,HLEC)共培養(yǎng)后,經(jīng)流式細(xì)胞儀、激光共聚焦顯微鏡及MTT法檢測二者進(jìn)入HLEC的能力及對HLEC的影響。體內(nèi)實(shí)驗(yàn)中,將GFP、R9-GFP分別通過滴眼的方式作用于小鼠眼球,7 d后裂隙燈下觀察后提取眼球總蛋白,ELISA法檢測眼球中重組蛋白的含量。結(jié)果 MTT結(jié)果顯示,R9-GFP、GFP都不影響HLEC的增殖活力。流式細(xì)胞儀和激光共聚焦顯微鏡檢測結(jié)果顯示,R9-GFP可以快速有效地進(jìn)入HLEC內(nèi)部且效果與給藥劑量呈正相關(guān)。體內(nèi)實(shí)驗(yàn)中,裂隙燈觀察結(jié)果示,GFP、R9-GFP作用后小鼠眼球均未發(fā)現(xiàn)異常;ELISA檢測結(jié)果顯示,R9-GFP可以進(jìn)入眼球內(nèi)部。結(jié)論 R9具有攜帶蛋白質(zhì)進(jìn)入細(xì)胞膜甚至穿透眼球屏障的能力。本研究為眼部用藥及給藥模式的進(jìn)一步改良提供了基礎(chǔ)。
[Abstract]:Objective to investigate whether polyarginine can penetrate the membrane and even the eyeball barrier with polyarginine as the carrier. Methods in this study, 9 polyarginine R9 was used as the transmembrane peptide. Two recombinant proteins of R9-green fluorescent protein (GFP) were constructed. In vitro, GFP was prepared. R9-GFP was co-cultured with human lens epithelial cell line HLEC. Laser confocal microscope and MTT method were used to detect the ability of HLEC entry and its effect on HLEC. In vivo, GFP R9-GFP was applied to mouse eyeball by dropping eyes. After 7 days observation under slit lamp, the content of recombinant protein in the eyeball was detected by Elisa. Results the results of MTT showed R9-GFP. GFP did not affect the proliferative activity of HLEC. The results of flow cytometry and laser confocal microscopy showed. R9-GFP could enter the HLEC quickly and effectively, and the effect was positively correlated with the dosage. In vivo, the slit lamp observation results showed that GFP. After R9-GFP treatment, no abnormal eyeballs were found in mice. The results of ELISA test showed that. Conclusion R9 has the ability to carry protein into the cell membrane and even penetrate the eyeball barrier. This study provides a basis for the further improvement of drug administration and administration pattern.
【作者單位】： 愛爾眼科醫(yī)院;
【基金】：黑龍江省衛(wèi)生計(jì)生委科研課題資助(編號:2014-044)~~
【分類號】：R77
【正文快照】： 隨著醫(yī)學(xué)及生物技術(shù)的發(fā)展,以抗體為代表的重組蛋白藥物越來越多地出現(xiàn)在眼科的臨床應(yīng)用中。由于角膜屏障及血-眼屏障的存在,分子量大、結(jié)構(gòu)復(fù)雜且滲透性較差的蛋白藥物不論是通過傳統(tǒng)的滴眼給藥還是全身注射,都不能有效到達(dá)眼球內(nèi)的病灶區(qū),因此臨床往往選擇玻璃體內(nèi)注射方式，

本文編號：1452249