小鼠伊文思蓝尾静脉或腹腔注射及视网膜铺片技术的优化  

Technique optimization of intravenous or intraperitoneal injection of Evans blue combined with retinal preparations in mice

作　　者：邢春涛 余琦 张璐莎 李珏 关华 王乐 李蓉 Chun-Tao Xing;Qi Yu;Lu-Sha Zhang;Jue Li;Hua Guan;Le Wang;Rong Li(School of Clinical Medicine,Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Shaanxi Key Laboratory of Ischemic Cardiovascular Diseases,Institute of Basic and Translational Medicine,Xi'an Medical University,Xi'an 710021,Shaanxi Province,China;Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi Province,China;Department of Ophthalmology,the First Affiliated Hospital of Xi'an Medical University,Xi'an 710077,Shaanxi Province,China)

机构地区：[1]宁夏医科大学临床医学院,中国宁夏回族自治区银川市750004 [2]西安医学院基础与转化医学研究所陕西省缺血性心血管疾病重点实验室,中国陕西省西安市710021 [3]陕西中医药大学,中国陕西省咸阳市712046 [4]西安医学院第一附属医院眼科,中国陕西省西安市710077

出　　处：《国际眼科杂志》2023年第7期1093-1098,共6页International Eye Science

基　　金：国家自然科学基金项目(No.81773795);陕西省科技资源开放共享平台资助项目(No.2020PT-003)。

摘　　要：目的:对小鼠伊文思蓝静脉推注联合视网膜铺片的技术进行优化,以提高视网膜铺片染色实验的准确性和可重复性。方法:C57BL/6雄性小鼠尾静脉注射10g/L(1%)伊文思蓝0.3mL后体内分别循环10、20min,处死后摘取眼球,4%多聚甲醛分别固定20、40、60min,将不同体内循环时间及不同组织固定时间对于视网膜血管的走形、分布及渗漏的影响进行了优化比较。尾静脉注射失败后,通过腹腔注射1%伊文思蓝0.3mL,体内循环3h,固定60min进行补救,观察视网膜血管的走形、分布及渗漏。同时将尾静脉注射后视网膜血管的走形、分布及渗漏与腹腔注射后的结果进行效果比较,确定最佳的体内循环时间以及视网膜铺片的条件。结果:尾静脉注射后,与体内循环20min,固定20或40min条件下视网膜血管情况相比,伊文思蓝体内循环10min,固定60min,视网膜血管走行、血管分支清楚,血管渗漏较少。尾静脉注射失败后,腹腔注射补救措施结果显示视网膜血管走行、各级血管形态清晰。腹腔注射伊文思蓝体内循环3h与尾静脉注射伊文思蓝体内循环10min相比,视网膜血管的走形、分布的效果无明显差别。结论:优化伊文思蓝静脉推注联合视网膜铺片的技术,可提高视网膜铺片染色实验的准确性和可重复性,为相关视网膜血管病变的研究提供借鉴方法。AIM:To optimize the technique of intravenous injection of Evans blue and retinal preparations in mice,improving the accuracy and repeatability of staining experiment of retinal preparations.METHODS:C57BL/6 male mice were intravenous injected with 10g/L (1%) Evans Blue 0.3mL and circulated in vivo for 10 or 20min,and the eyes were removed after sacrificed and fixed in 4% paraformaldehyde for 20,40 or 60min.When failure of intravenous injection,the experiment was remediated by intraperitoneal injection of 1% Evans Blue 0.3mL,circulated in vivo for 3h and fixed for 60min to observe morphology,distribution and leakage of the retinal vessels.Besides,we compared the morphology,distribution and leakage of the retinal vessels after intravenous injection with those after intraperitoneal injection to determine the optimal conditions for in vivo circulation time and retinal preparations.RESULTS:After intravenous injection,compared to the retinal vascular condition under 20min in vivo circulation time of Evans blue and 20 or 40min of fixation,with 10min of in vivo Evans blue circulation and 60min of fixation,the morphology of retinal vascular was more intact with less retinal vascular leakage,and the vascular branches are clear.When intravenous injection failed,remediated results from intraperitoneal injection showed that the morphology and distribution of retinal vessels were intact.There was no significant difference in morphology,distribution and leakage of the retinal vessels after 3h of intraperitoneal Evans blue circulation compared to 10min intravenous Evans blue circulation.CONCLUSION:This experiment optimizes the protocol,improves the accuracy and reproducibility of retinal preparations,and provides a reference for the study of related retinal vascular diseases.

关 键 词：视网膜铺片 伊文思蓝 血管造影 眼球固定 静脉注射 腹腔注射 

分 类 号：R774.1[医药卫生—眼科] R-332[医药卫生—临床医学]