针刺对干眼兔角膜形态学及角膜组织NF-κB信号通路的影响  

Effect of acupuncture on corneal morphology and nuclear factor κB signaling pathway of dry eye rabbits

作　　者：丁宁[1] 韦庆波 刘成勇[2] 邓伟民[3] 徐倩[1] 高卫萍[2] DING Ning;WEI Qingbo;LIU Chengyong;DENG Weimin;XU Qing;GAO Weiping(Nanjing University of Chinese Medicine,Nanjing 210008,Jiangsu Province,China;Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,Jiangsu Province,China;Zhongda Hospital,Southeast University,Nanjing 210029,Jiangsu Province,China)

机构地区：[1]南京中医药大学,江苏省南京市210008 [2]南京中医药大学附属医院,江苏省南京市210029 [3]东南大学附属中大医院,江苏省南京市210029

出　　处：《眼科新进展》2024年第3期173-177,共5页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:82074526,82305377,82205259);江苏省自然科学基金青年基金资助(编号:BK20230455)。

摘　　要：目的 探讨针刺对干眼兔角膜形态学及角膜组织NF-κB信号通路的影响,分析针刺对干眼的作用机制。方法 取雌雄不拘的健康新西兰兔24只,随机分为空白组、模型组、针刺组、假针刺组,每组6只。空白组为正常对照组,不加任何处理;其余3组动物每天800、 1100、1400和1800时于皮下注射氢溴酸东莨菪碱2.0 mg·kg-1,连续35 d直至实验结束。假针刺组:于造模后第22天给予假针刺治疗(睛明BL1、攒竹BL2、丝竹空SJ23、太阳穴EX-HN5、瞳子髎GB1),钝针头点刺穴位,不刺进穴位,每天1次,连续14 d。针刺组:于造模成功后第22天给予针刺治疗,穴位同假针刺组。造模后第0、21、28、35天分别进行角膜荧光素染色,第35天进行角膜共焦显微镜检查,之后处死动物,在光学显微镜、透射电子显微镜下观察角膜形态学变化,采用Western blot检测角膜组织NF-κB蛋白的表达。结果 与模型组相比,造模后第28、35天,针刺组、空白组兔角膜荧光素染色评分均减小,差异均有统计学意义(均为P<0.05)。造模后第35天共焦显微镜检查结果显示,模型组和假针刺组与其他组相比,兔角膜基质层出现大量球状免疫细胞和边界不清大小不规律的活化基质层细胞,可见具有不规则细胞间间隙的区域,存在炎症;针刺组基质层细胞形态得到改善,细胞呈轻微激活状态,角膜神经形态未见明显异常。造模后第35天光学显微镜检查结果显示,模型组和假针刺组兔角膜组织表面可见角化过度的扁平上皮细胞,淋巴细胞浸润,局灶上皮细胞层数增多,表面上皮细胞脱落;针刺组角膜上皮有接近4~6层上皮细胞,上皮脱落减少,与模型组相比,淋巴细胞浸润减少。造模后第35天透射电子显微镜检查结果显示,模型组和假针刺组兔角膜上皮细胞出现异常的微绒毛结构和上皮细胞缺失,细胞间隙增宽,粗面内质网严重扩张,桥粒解体伴随线粒体肿胀;针刺组上�Objective To investigate the effect of acupuncture on the morphology of the dry eye rabbit’s cornea and the nuclear factorκB(NF-κB)signaling pathway of the corneal tissue to analyze the mechanism of acupuncture on dry eyes.Methods Twenty-four healthy New Zealand rabbits,without restriction on sex,were randomly divided into four groups,including a blank group,a model group,an acupuncture group,and a sham acupuncture group,with 6 in each group.Rabbits in the blank group were not treated;rabbits in the other three groups were treated with scopolamine hydrobromide 2.0 mg·kg-1 by subcutaneous injection at 8 GA6FA 00,11 GA6FA 00,14 GA6FA 00 and 18 GA6FA 00 each day for 35 consecutive days until the end of the experiment.Rabbits in the sham acupuncture group were treated with sham acupuncture on the 22nd day after successful modeling by quickly pricking acupoints(Jingming BL1,Cuanzhu BL2,Sizhukong SJ23,Taiyang EX-HN5 and Tongziliao GB1)with a blunt acupuncture needle,once a day,for a total of 14 days.Rabbits in the acupuncture group were treated with acupuncture at the same acupoints as the sham acupuncture group after successful modeling.The corneal fluorescence staining was conducted on Days 0,21,28 and 35 after modeling.On Day 35,corneal confocal microscope examinations were conducted.Then,the rabbits were sacrificed,the corneal morphological changes were observed by light microscope and transmission electron microscope,and the expression of corneal NF-κB protein was detected by Western blot.Results Compared with the model group,the score of rabbit corneal fluorescein staining in the acupuncture group and blank group decreased on the 28th and 35th days after modeling,and the differences were statistically significant(all P<0.05).The results of the confocal microscope examination on Day 35 after modeling showed that,compared with other groups,there were a large number of globular immune cells and activated stromal cells with unclear boundaries and irregular sizes in the stromal layer and inflammation in the area

关 键 词：针刺 干眼 角膜形态学 NF-ΚB信号通路 

分 类 号：R772.2[医药卫生—眼科]