CAJ | 학술논문

目的探讨三七皂苷(PNS)对大鼠持续性高眼压下视网膜神经节细胞损伤的保护作用及机制.方法健康SD大鼠80只随机分为四组,正常对照组、0.9%氯化钠溶液组、单纯用药组和联合用药组.采用烙闭巩膜上静脉联合术后结膜下注射5-FU(5-氟尿嘧啶)的方法制作大鼠持续性高眼压模型,所有大鼠定期测量并记录眼压.4周、8周、12周、16周、20周后分别处死大鼠,摘取眼球,TUNEL法检测视网膜神经细胞(Retinal ganglion Cells,RGCs)凋亡;AgNOR染色检测RGCs的活性;免疫组织化学检测视网膜节细胞层半胱氨酸蛋白酶-9(caspase-9)的表达.结果术后眼压维持在(36.55±2.27)mm Hg为4周至20周左右;视网膜神经节细胞层TUNEL阳性细胞数与正常对照组差异有统计学意义;正常对照组和联合用药组RGC8细胞核内银染颗粒数比较差异无统计学意义(P>0.05);各组视网膜神经节细胞层caspase-9蛋白的表达较正常对照组明显增强.结论PNS对持续性高眼压导致的视网膜神经节细胞损伤有较显著保护作用.联合降眼压药控制眼压,PNS对大鼠视网膜神经节细胞的保护作用更突出.PNS能抑制视网膜神经节细胞Caspase-9的表达从而保护大鼠视网膜神经节细胞.
목적 탐토삼칠조감(PNS)대대서지속성고안압하시망막신경절세포손상적보호작용급궤제.방법 건강SD대서80지수궤분위사조,정상대조조、0.9%록화납용액조、단순용약조화연합용약조.채용락폐공막상정맥연합술후결막하주사5-FU(5-불뇨밀정)적방법제작대서지속성고안압모형,소유대서정기측량병기록안압.4주、8주、12주、16주、20주후분별처사대서,적취안구,TUNEL법검측시망막신경세포(Retinal ganglion Cells,RGCs)조망;AgNOR염색검측RGCs적활성;면역조직화학검측시망막절세포층반광안산단백매-9(caspase-9)적표체.결과 술후안압유지재(36.55±2.27)mm Hg위4주지20주좌우;시망막신경절세포층TUNEL양성세포수여정상대조조차이유통계학의의;정상대조조화연합용약조RGC8세포핵내은염과립수비교차이무통계학의의(P>0.05);각조시망막신경절세포층caspase-9단백적표체교정상대조조명현증강.결론PNS대지속성고안압도치적시망막신경절세포손상유교현저보호작용.연합강안압약공제안압,PNS대대서시망막신경절세포적보호작용경돌출.PNS능억제시망막신경절세포Caspase-9적표체종이보호대서시망막신경절세포.
Objective To investigate protective effects and mechanisms of extract of PNS on retinal ganglion cells injury induced by continuous high intraocular pressure (IOP) in rats.Methods 80 healthy Sprague-Dawley ( SD) rats were randomly divided into four groups to establish rat model of high intraocular pressure with 4,8,12,16,20 weeks which there were cauterizing episcleral veins combined 5-Fu and only cauterizing episcleral veins.All the rats intraocular pressure was measured and recorded regularly.After 4,8,12,16,20 weeks,all rats were killed and the eyeballs were removed and to assay apoptosis of RGCs by TUNEL,to detect the activity of RGCs AgNOR staining and to discover the expression of caspase-9 by immunohistochemical detections.Results The IOP was almostly more than 36.55±0.27mmHg.The order of the number of TUNEL-positive cells in retinal ganglion cell layer,compared with the normal control group,there was a significant difference.Stained grains there was no significant difference between the combined treatment group and normal control group (P>0.05).The expression of caspase-9 protein in the saline group,treatment group and combined treatment group was obviously enhanced according to the normal control group.Conclusion The sustainable and stable rat model of high intraocular pressure could be established by cauterization of episcleral veins whih subconjunctival injection 5-Fu.PNS had significant protective effects in RCCs injury caused by the persistent high intraocular pressure.If controlling intraocular pressure with drugs which could lower the IOP,the protective effects of PNS on RGCs would be more prominent.PNS could inhibit the expression of Caspase-9 in the rat RGCs to protect RGCs.