基于miR-146a/JNK1通路探究益气针法治疗混合型干眼大鼠模型机制研究  

Exploring the Anti-Inflammatory Effects of Qi Tonifying Acupuncture Method in a Mixed-Type Dry Eye Rat Model through the miR-146a/JNK1 Pathway

作　　者：赵磊[1] 董宝强[2] 左韬[1] 刘建宇 姜艳华[3] 张静涵 Zhao Lei;Dong Baoqiang;Zuo Tao;Liu Jianyu;Jiang Yanhua;Zhang Jinghan(Department of Ophthalmology,the Second Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110034,China;Liaoning University of Traditional Chinese Medicine,Shenyang 110032,China;Department of Ophthalmology,the Fourth People's Hospital of Shenyang,China Medical University,Shenyang 110031,China)

机构地区：[1]辽宁中医药大学附属第二医院眼科,沈阳110034 [2]辽宁中医药大学,沈阳110032 [3]中国医科大学沈阳市第四人民医院眼科,沈阳110031

出　　处：《中华眼视光学与视觉科学杂志》2025年第2期88-95,共8页Chinese Journal Of Optometry Ophthalmology And Visual Science

基　　金：国家自然科学基金(82104714);中国博士后科学基金面上项目(2023MD734185);辽宁省科学技术基金博士科研启动基金(2022-BS-069);辽宁省教育厅青年项目(LJKQZ20222404)。

摘　　要：目的:研究益气针法基于微小核糖核酸-146a(miR-146a)/c-Jun氨基末端激酶1(JNK1)信号通路防治混合型干眼大鼠模型的作用机制。方法:实验研究。2022年6月将50只Wistar大鼠随机分为空白组、模型组、针刺组、agomiR-146a组和针刺联合agomiR-146a组,每组10只。空白组不做任何处理,其余各组大鼠采用固视联合吹风诱导混合型干眼模型后,模型组不予处理,针刺组予太阳、攒竹、四白、期门、合谷、气海、天宗、风池、颈夹脊针刺治疗;agomiR-146a组予注射miR-146a激动剂agomiR-146a;针刺联合agomiR-146a组给予针刺组和agomiR-146a组干预措施联合应用。末次干预后观察比较各组大鼠泪膜破裂时间(BUT)、角膜荧光素染色(CFS);观察比较各组大鼠角膜上皮细胞的形态及超微结构,实时定量PCR检测miR-146a、基质金属蛋白酶-9(MMP-9)、白细胞介素-1β(IL-1β)、含半胱氨酸的天冬氨酸蛋白水解酶-3(caspase-3)的mRNA表达;Western Blot检测磷酸化-JNK1(p-JNK1)、JNK1、MMP-9、IL-1β、caspase-3的蛋白表达。多组间数据比较采用单因素方差分析,两两比较采用LSD-t检验。结果:与空白组比较,模型组大鼠BUT水平明显降低(P<0.001),CFS评分明显增高(P<0.001),角膜组织中miR-146a的mRNA相对表达量明显降低(P<0.001),p-JNK1/JNK1的蛋白表达明显增高(P<0.001),MMP-9、IL-1β、caspase-3的mRNA及蛋白表达均明显增加(均P<0.01)。与模型组比较,各干预组的BUT均明显增加(均P<0.01),CFS评分均明显降低(均P<0.01),角膜组织中miR-146a的mRNA相对表达量明显增加(均P<0.01),p-JNK1/JNK1的蛋白表达明显降低(P<0.001),MMP-9、IL-1β、caspase-3的mRNA及蛋白表达均明显降低(均P<0.01)。与针刺组比较,agomiR-146a组、针刺联合agomiR-146a组BUT水平增高(P=0.013、0.001),CFS评分降低(P=0.003,P<0.001),角膜组织中miR-146a的mRNA相对表达量均增加(均P<0.001),p-JNK1/JNK1的蛋白表达均降低(均P<0.001),MMP-9、IL-1β、caspase-3的mRNObjective:To investigate the effects of Qi tonifying acupuncture in a rat model of mixed-type dry eye,focusing on the intricate micro RNA-146a/c-Jun N-terminal kinase 1(miR-146a/JNK1)pathway.Methods:This was an experimental study.In June 2022,Fifty Wistar rats were randomly divided into blank group,model group,acupuncture group,agomiR-146a group,and acupuncture combined with agomiR-146a group,with 10 rats in each group.Adopting a fixed vision combined with wind blowing to induce a mixed-type dry eye rat model.And the acupuncture group was treated with Taiyang,Zanzhu,Sibai,et al acupuncture.The agomiR-146a group was injected with the miR-146a agonist agomiR-146a.The acupuncture combined with agomiR-146a group was administered with a combination of acupuncture and agomiR-146a intervention measures.The break up time(BUT)and corneal fluorescence staining(CFS)after the last intervention was observed.The morphology and ultrastructure of corneal epithelial cells were also observed.And the mRNA and protein expression of the miR-146a,phospho-C-Jun N-terminal kinase 1(p-JNK1),c-jun N-terminal kinase 1(JNK1),matrix metallo proteinase-9(MMP-9),interlukin-1β(IL-1β),and cysteinyl aspartate specific proteinase-3(caspase-3)were detected in this study as well.One way ANOVA analysis was used for inter-group comparison,and LSD-t test was used for pairwise comparison.Results:Compared with the blank group,the BUT level of the model group was significantly reduced(P<0.001),the CFS score was significantly increased(P<0.001),the relative mRNA expression of miR-146a was significantly reduced(P<0.001),and the protein expression of p-JNK1/JNK1 was significantly increased(P<0.001),as well as the mRNA and protein expression of MMP-9,IL-1β,and caspase-3 were significantly increased(P<0.01).Compared with the model group,the BUT of each intervention group was significantly increased(P<0.01),the CFS score was significantly decreased(P<0.01),the relative mRNA expression of miR-146a was significantly increased(P<0.01),and the protein expression

关 键 词：干眼症 微小核糖核酸-146a c-Jun氨基末端激酶1 针刺 角膜 大鼠 

分 类 号：R246.82[医药卫生—针灸推拿学] R246.81[医药卫生—中医临床基础]