基于PEDF-JAK2/STAT3通路探讨电针对视网膜缺血再灌注所致视网膜神经节细胞凋亡的抑制作用  

Exploration of inhibitory effect of electroacupuncture on retinal ganglion cells apoptosis induced by retinal ischemia reperfusion based on PEDF-JAK2/STAT3 pathway

作　　者：符甜甜 吴文艳 陈平 郭润杰 徐红[2] 张仁[3] 田雪松 FU Tiantian;WU Wenyan;CHEN Ping;GUO Runjie;XU Hong;ZHANG Ren;TIAN Xuesong(Experiment Center for Science and Technology,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Department of Acupuncture-Moxibustion,Longhua Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China;Shanhai Chinese Medicine Literature Museum,Shanghai 200025,China)

机构地区：[1]上海中医药大学科技实验中心,上海201203 [2]上海中医药大学附属龙华医院,上海200032 [3]上海市中医文献馆,上海200025

出　　处：《上海中医药大学学报》2024年第5期44-52,共9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);上海市金山区医学重点专科建设项目(JSZK2023H05)。

摘　　要：目的:探讨电针“睛明”“新明”两穴对视网膜缺血再灌注损伤(RIRI)的保护作用,并阐明相关机制。方法:Wistar大鼠高眼压(HIOP)模型模拟RIRI。HIOP 12 h组在造模结束后立即电针“睛明”“新明”两穴30 min,HIOP 24 h组在造模结束后及再灌注12 h分别电针“睛明”“新明”两穴30 min。Western blot检测Janus激酶信号转导子与转录激活子(JAK/STAT)激活,苏木精-伊红染色法(HE)观察视网膜厚度及视网膜神经节细胞(RGCs)细胞数目,末端脱氧核苷酸转移酶dUTP缺口末端标记(TUNEL)染色观察RGCs凋亡,酶联免疫吸附测定(ELISA)法检测视网膜色素上皮衍生因子(PEDF)的含量。结果:p-STAT3在RIRI 6 h开始升高(P<0.05),12 h达到顶峰(P<0.01),24 h逐渐下降(P<0.01),p-JAK2蛋白表达水平与p-STAT3大致呈现相同的趋势;RIRI 12 h时电针组p-STAT3/STAT3、p-JAK2/JAK2表达水平显著下降(P<0.05,P<0.01);RIRI 12 h时电针组PEDF含量明显高于HIOP组(P<0.001);HE染色结果显示,电针组RGCs数目、视网膜总厚度、GCL层、INL(inner nuclear layer)层厚度与HIOP组相比,均显著增加(P<0.01,P<0.001);TUNEL染色显示,电针组RGCs凋亡数量与HIOP组相比明显减少(P<0.05,P<0.01)。结论:电针可能通过提高PEDF水平,抑制JAK2/STAT3通路,从而抑制RGCs凋亡。Objective:To explore the protective effect of electroacupuncture(EA)at"Jingming"and"Xinming"acupoints on retinal ischemia-reperfusion injury(RIRI)and clarify the related mechanism.Methods:Wistar rats with high intraocular pressure(HIOP)were modeled to simulate RIRI.The HIOP 12 h group was treated with electroacupuncture at"Jingming"and"Xinming"for 30 minutes immediately after modeling,and the HIOP 24 h group was treated with electroacupuncture at"Jingming"and"Xinming"for 30 minutes respectively after modeling and reperfusion for 12 h.Western blot was used to detect the activation of Janus kinase signal transducer and activator of transcription(JAK/STAT).The retinal thickness and the number of retinal ganglion cells(RGCs)were observed by hematoxylin-eosin staining(HE).The apoptosis of RGCs was observed by TdT-mediated dUTP Nick-End Labeling(TUNEL)staining.The content of retinal pigment epithelium-derived factor(PEDF)was detected by ELISA.Results:p-STAT3 began to increase at 6 h after RIRI(P<0.05),peaked at 12 h(P<0.01),and gradually decreased at 24 h(P<0.01).The expression level of p-JAK2 protein showed a similar trend as that of p-STAT3.The expression levels of p-STAT3/STAT3 and p-JAK2/JAK2 in the RIRI 12 h EA group were significantly decreased(P<0.05,P<0.01).The content of PEDF in RIRI 12 h EA group was significantly higher than that in HIOP group(P<0.001).The results of HE staining showed that the number of RGCs,total retinal thickness,GCL layer and inner nuclear layer(INL)layer thickness in the EA group were significantly increased compared with those in the HIOP group(P<0.01,P<0.001).TUNEL staining showed that the number of apoptotic RGCs in the EA group was significantly reduced than that in the HIOP group(P<0.05,P<0.01).Conclusion:Electroacupuncture may inhibit RGCs apoptosis by increasing PEDF level and inhibiting JAK2/STAT3 pathway.

关 键 词：电针 JAK2/STAT3通路 视网膜缺血再灌注损伤 神经节细胞 凋亡 

分 类 号：R245[医药卫生—针灸推拿学]