δ-阿片受体在电针抗视网膜缺血再灌注致视网膜细胞坏死性凋亡中的作用  

Role of δ-opioid receptors in electroacupuncture against retinal ischemia-reperfusion induced retinal cell necroptosis

作　　者：郭润杰 陈平 符甜甜 陈静 夏勇[3] 张仁[4] 徐红[5] 田雪松 GUO Runjie;CHEN Ping;FU Tiantian;CHEN Jing;XIA Yong;ZHANG Ren;XU Hong;TIAN Xuesong(Experiment Center for Science and Technology,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shanghai Jinshan TCM-Integrated Hospital,Shanghai 201501,China;School of Acupuncture-Moxibustion and Tuina,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shanghai Literature Institute of Traditional Chinese Medicine,Shanghai 200025,China;Department of Acupuncture-Moxibustion,Longhua Hospital Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学科技实验中心,上海201203 [2]上海市金山区中西医结合医院,上海201501 [3]上海中医药大学针灸推拿学院,上海201203 [4]上海市中医文献馆,上海200025 [5]上海中医药大学附属龙华医院针灸科,上海201203

出　　处：《上海中医药大学学报》2022年第5期52-59,共8页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81574078,81072646);上海市进一步加快中医药事业发展三年行动计划项目(2018年~2020年);上海市金山区第六周期医学重点专科建设项目(JSZK2019H04)。

摘　　要：目的:探讨δ-阿片受体(DOR)在电针抗高眼压(HIOP)诱导视网膜缺血再灌注损伤致视网膜细胞坏死性凋亡中的作用。方法:55只Wistar大鼠随机分为5组:空白对照组、HIOP 6 h组、HIOP 12 h组、HIOP 24 h组、HIOP72 h组,PI染色检测上述时间点大鼠视网膜细胞坏死性凋亡数量,Western blot检测p-Rip1、p-Rip3表达。另取55只Wistar大鼠随机分为5组:空白对照组、HIOP组、电针+HIOP组、假电针+HIOP组、DOR拮抗剂+HIOP+电针组,除空白对照组外,其余各组均采用HIOP法致大鼠视网膜缺血再灌注,缺血1 h再灌24 h。DOR拮抗剂+HIOP+电针组缺血前1 h玻璃体腔注射DOR拮抗剂Naltrindole,再灌注开始及再灌注12 h,电针“水沟“”睛明”穴两次,每次30 min,再灌注24 h,PI染色检测视网膜中坏死性凋亡细胞数量,Western blot检测p-Rip1、p-Rip3表达。结果:HIOP 6、12、24、72 h,视网膜坏死性凋亡细胞数量明显增多,在12 h达到高峰,视网膜中p-Rip1和p-Rip3蛋白表达亦显著增加,其中p-Rip1峰值出现在12 h,p-Rip3在24 h表达最高;再灌注24 h,电针+HIOP组坏死性凋亡细胞数量、p-Rip1和p-Rip3蛋白表达均显著低于HIOP组,假电针+HIOP组和DOR拮抗剂+HIOP+电针组与HIOP组比较,坏死性凋亡细胞数量、p-Rip1和p-Rip3蛋白表达均无显著性差异。结论:在HIOP法诱导的视网膜缺血再灌注损伤中,电针“水沟”“睛明”两穴可显著减少视网膜细胞坏死性凋亡,对视网膜具有保护作用,且电针的抗视网膜细胞坏死性凋亡作用可能和DOR密切相关。Objective:To investigate the role of delta-opioid receptor(DOR) in electroacupuncture(EA) against retinal cell necroptosis caused by retinal ischemia-reperfusion induced by high intraocular pressure(HIOP).Methods:Fifty-five Wistar rats were randomly divided into five groups:blank control group,HIOP 6 h group,HIOP 12 h group,HIOP 24 h group and HIOP 72 h group. PI staining was used to detect the number of necroptotic cells in rats’ retina at the above timings,and Western blot was used to detect the expressions of p-Rip1 and p-Rip3. Another fifty-five Wistar rats were randomly divided into 5 groups:blank control group,HIOP group,EA+HIOP group,sham EA+HIOP group,and DOR antagonist+HIOP+EA group. Except for the blank control group,HIOP was used in the other groups to induce retinal ischemia-reperfusion in rats,and the ischemia lasted for 1 h,then reperfused for 24 h. DOR antagonist+HIOP+EA group received intravitreal injection of DOR antagonist Naltrindole 1 h before ischemia. At the beginning of reperfusion and 12 h after reperfusion,EA was performed at acupoints“Shuigou”and“Jingming”twice,30 min each time. 24 h after reperfusion,the number of necroptotic cells in retina was detected by PI staining,and the expressions of p-Rip1 and p-Rip3 were detected by Western blot.Result:After HIOP for 6 h,12 h,24 h and 72 h,the number of retinal necroptotic cells was increased significantly,and reached a peak at 12 h.The protein expressions of p-Rip1 and p-Rip3 in the retina were also significantly increased,and the peak of p-Rip1 expression appeared at12 h and the expression of p-Rip3 was the highest at 24 h. After 24 h of reperfusion,the number of necroptotic cells and the protein expressions of p-Rip1 and p-Rip3 in the EA+HIOP group were all significantly lower than those in the HIOP group. Compared with the HIOP group,there was no significant difference in the number of necroptotic cells or the protein expressions of p-Rip1 and p-Rip3 in the sham EA+HIOP group and the DOR antagonist+HIOP+EA group.Conclusion:In ret

关 键 词：电针 视网膜缺血再灌注损伤 Δ阿片受体 坏死性凋亡 

分 类 号：R774.1[医药卫生—眼科]