冰片对体外培养大鼠视网膜血管内皮细胞屏障通透性的影响  

Effect of borneol on permeability of the blood retinal barrier of cultured vascular endothelial cells in rats in vitro

作　　者：闫磊[1] 桂金秋[1] 刘志新[1] 孙晓冬[1] 梁军[1] 邵新然 蔡克瑞[1] YAN Lei;GUI Jin - qiu;LIU Zhi - xin;SUN Xiao - dong;LIANG Jun;SHAO Xin -ran;CAI Ke - ru(Department of Histology and Embryology, Mudanjiang Medical University, Mudanjiang 157011, Heilongjiang Province, China)

机构地区：[1]牡丹江医学院组胚教研室,黑龙江牡丹江157011 [2]牡丹江医学院

出　　处：《中国临床药理学杂志》2018年第11期1371-1374,共4页The Chinese Journal of Clinical Pharmacology

基　　金：黑龙江省卫生计生委科研课题基金资助项目(2017-312); 黑龙江省教育厅科学技术研究基金资助项目(11551529); 黑龙江省大学生创新创业训练计划重点基金资助项目(201710229006); 牡丹江市科学技术计划基金资助项目(Z2016s0088)

摘　　要：目的研究冰片对体外培养大鼠血视网膜屏障通透性影响及其作用机制。方法获取大鼠视网膜血管内皮细胞(REVC),经原代培养并鉴定。大鼠REVC经胰酶消化后接种于铺有层黏连蛋白的Transwell小室中,培养15 d后成功建立REVC紧密连接屏障模型。将紧密连接屏障模型分为2组:实验组(加入冰片血清)和对照组(加入空白血清),用跨上皮细胞电阻测量仪测定2组在0,12,24h这3个时间点的跨上皮细胞电阻(TER)值;以逆转录-聚合酶链反应法检测2组的紧密连接相关蛋白ZO-1 mRNA表达水平。结果在12 h的TER值:实验组与对照组分别为(159.24±3.51),(269.53±5.86)Ω×cm^2,组间比较差异有统计学意义(P<0.01);但在24 h,组间差异无统计学意义(P>0.05),表明加入冰片血清12 h后,大鼠REVC间紧密连接间隙增大,紧密连接开放,导致TER值降低,24 h后紧密连接恢复正常,TER值恢复到0 h水平,表明冰片具有开放体外培养的大鼠REVC间紧密连接的作用。在12 h的ZO-1 mRNA值:实验组与对照组分别为0.47±0.39,0.80±0.02,组间比较差异有统计学意义(P<0.01);但在24 h,组间差异无统计学意义(P>0.05),表明加入冰片后,大鼠REVC的紧密连接相关蛋白ZO-1 mRNA值表达减少,而后逐渐增加,在24 h恢复到0 h的水平。表明冰片可以下调大鼠REVC的紧密连接相关蛋白ZO-1mRNA表达水平,降低紧密连接相关蛋白ZO-1蛋白的表达。结论冰片可在转录水平上减少紧密连接相关蛋白ZO-1 mRNA表达,能够开放大鼠REVC间的紧密连接,增加血视网膜屏障的通透性。Objective To observe the effect of borneol on the permeability of rat blood retinal barrier in vitro,and explore the mechanism of borneol on the impact of tight junctions between cultured retinal vascular endothelial cells（ REVC） in rats. Methods REVC in rats was obtained,cultured and identified in primary culture. REVC was digested with trypsin and seeded in Transwell chamber with laminin,Transwell chambers were used to establish tight junctions barrier model between cultured REVC in rats after 15 d. There were 2 groups： the experimental group with the addition of borneol serum and the control group with blank serum. The borneol through transmembrance electric resistance（ TER）were determined at the time of 0,12,24 h after administration by Cross epithelial cell resistance measuring instrument. The tight junction associated protein ZO-1 mRNA expression were detected by reverse transcription polymerase chain reaction（ RT-PCR）. Results The TER＇s values at 12 h in the experimental group and the control group were（ 159. 24 ±3. 51）,（ 269. 53 ± 5. 86） Ω × cm^2,the difference was significant（ P〈0. 01）; but the difference at the 24 h had no significant difference（ P〈0. 05）. It was indicated that the gap of tight junctions increased,tight junctions was opened and TER decreased after the addition of borneol serum for 12 h. After 24 h,tight junctions returned to normal state and TER was returned to the 0 h level; it showed that borneol could open tight junctions between REVC of rats in vitro. The ZO-1 mRNA＇s values in experimental group and control group were 0. 47 ± 0. 39,0. 80 ± 0. 02 at 12 h. The difference in 2 groups was statistically significant（ P〈0. 01）; but the difference in 2 groups was not statistically significant at the 24 h（ P〉0. 05）. ZO-1 mRNA had the similar tendency with TER. Conclusion Borneol could decrease the expressions of tight junction associated proteins ZO-1,which can open tight junctions between REVC in rats and increase tight junctions perme

关 键 词：冰片 视网膜血管内皮细胞 通透性 视网膜屏障 

分 类 号：R97[医药卫生—药品]