摘要
目的探索一种旨在阻断干眼形成通路且具有抗炎、缓释和良好生物相容性等特性的纳米药物。方法采用薄膜分散-水化超声法制备汉防己甲素(Tet)和聚乳酸-羟基乙酸共聚物(PLGA)的纳米微球(Tet-ATS@PLGA)并检测其室温(25℃)下稳定性、包封率、载药量等性质。分组实验:正常组(未行干预的正常兔眼);制备成功的干眼模型兔随机分为控制组(未行任何干预)、ATS组(人工泪液干预)、Tet-ATS组(人工泪液及Tet干预)、Tet-ATS@PLGA组(Tet-ATS@PLGA干预)。流式细胞术检测各组干预24 h后炎症化兔角膜上皮细胞凋亡率;干预14 d后,观察并记录兔角膜上皮细胞着染情况、泪膜破裂时间(BUT)、眼表泪液分泌(Schirmer试纸检测)情况;HE染色观察兔角膜上皮细胞厚度、球结膜杯状细胞形态和数量;提取兔角膜蛋白行ELISA实验检测血管内皮生长因子(VEGF)、白细胞介素(IL)-1β、前列腺素E2(PGE2)和肿瘤坏死因子(TNF)-α等炎症因子表达情况。采用独立样本t检验进行组间比较。结果Tet-ATS@PLGA纳米药物的包封率和载药量分别为77.43%和30.26%,室温下性质稳定,在眼表温度(33℃)下易释放。与其他组比较,Tet-ATS@PLGA组BUT最大,泪液分泌量最大,角膜上皮厚度恢复接近正常,球结膜杯状细胞数量恢复最多,24 h后炎症化兔角膜上皮细胞细胞凋亡率最高,VEGF、IL-1β、TNF-α和PGE2表达水平最低,差异均有统计学意义(均为P<0.05)。结论Tet-ATS@PLGA纳米药物可有效作用于炎症化兔角膜上皮细胞,促进炎症细胞凋亡,并进一步通过抑制VEGF、IL-1β、TNF-α和PGE2等炎症因子的表达,阻断干眼的炎症反应,改善眼表泪液分泌情况。
Objective To discover a nano-drug with anti-inflammatory,sustained-release,and biocompatible properties aimed at blocking the dry eye formation pathway.Methods Nano-microspheres(Tet-ATS@PLGA)composed of tetrandrine(Tet)and poly(lactic-co-glycolic acid)(PLGA)were prepared using the thin-film hydration method,and their stability at room temperature(25℃),encapsulation efficiency,and drug loading were tested.Normal rabbit eyes without intervention were recruited in the normal group,and dry eye models were randomly divided into the control group(without any intervention),ATS group(intervened by artificial tears),Tet-ATS group(intervened by artificial tears and Tet),and Tet-ATS@PLGA group(intervened by Tet-ATS@PLGA).Flow cytometry was performed to detect the apoptosis of inflammatory corneal epithelial cells in each group after 24 hours of intervention.The staining of corneal epithelial cells,tear film break-up time(BUT),and surface tear secretion(detected by the Schirmer test strip)were recorded after 14 days of intervention.The thickness of corneal epithelial cells as well as the shape and number of bulbar conjunctival goblet cells were monitored by hematoxylin-eosin staining.Corneal proteins were extracted for the enzyme-linked immunosorbent assay to measure the expression levels of vascular endothelial growth factor(VEGF),interleukin-1β(IL-1β),prostaglandin E2(PGE2),and tumor necrosis factor-α(TNF-α).The independent samples t-test was carried out for comparison among groups.Results The encapsulation efficiency and drug loading of Tet-ATS@PLGA nano-drug were 77.43% and 30.26%,respectively.The drug was stable at room temperature and easy to release when the ocular surface temperature stood at 33℃.Compared with other groups,BUT and the amount of tear secretion in the Tet-ATS@PLGA group were the largest,the thickness of corneal epithelial cells was close to the normal value,bulbar conjunctival goblet cells recovered the most,the apoptosis of inflammatory corneal epithelial cells after 24 hours of intervention was the most significant,and the expression levels of VEGF,IL-1β,TNF-α,and PGE2 were the lowest(all P<0.05).Conclusion Tet-ATS@PLGA nano-drug can effectively act on inflammatory corneal epithelial cells in rabbits,promote apoptosis of inflammatory cells,and block the inflammatory response of dry eyes by inhibiting the expression of VEGF,IL-1β,TNF-α,and PGE2,thus improving tear secretion on the ocular surface.
作者
李涛
张玉
朱胜兰
唐娟
刘兴德
吴小利
方其林
李盈
漆星
林胜
戴传强
周燕
LI Tao;ZHANG Yu;ZHU Shenglan;TANG Juan;LIU Xingde;WU Xiaoli;FANG Qilin;LI Ying;QI Xing;LIN Sheng;DAI Chuanqiang;ZHOU Yan(West China Hospital Sichuan University Ziyang Hospital(the First People’s Hospital of Ziyang),Ziyang 641300,Sichuan Province,China;Chongqing Nanping Aier Eye Hospital,Chongqing 400020,China;Ziyang College of Dental Technology,Ziyang 641300,Sichuan Province,China)
出处
《眼科新进展》
CAS
北大核心
2023年第9期697-702,共6页
Recent Advances in Ophthalmology
基金
四川省资阳市科技计划项目(编号:zykjjsc20-cxpt-2020-02,zykjjsc20-yyjc-2023-04)。
作者简介
李涛(ORCID:0000-0003-0816-7473),男,1990年3月出生,四川资阳人,硕士研究生,重庆医科大学眼科学专业在读博士,主治医师。研究方向:眼底病和白内障。E-mail:1138583262@qq.com;通信作者:张玉(ORCID:0009-0004-6582-6353),女,1987年1月出生,重庆人,硕士研究生,主治医师。研究方向:白内障和纳米材料。E-mail:517340898@qq.com。
