摘要
目的观察电针对干眼大鼠角膜自噬的调节作用。方法将雄性Sprague-Dawley(SD)大鼠随机分为正常组、模型组、电针组和假针组。除正常组外,其余3组均采用皮下注射东莨菪碱溶液配合吹风制备大鼠干眼模型。电针组和假针组造模后分别予电针及假针刺干预。检测大鼠干预前后泪膜破裂时间和泪液分泌量,并评估角膜荧光素染色的分值;应用透射电镜观察大鼠角膜组织的超微结构;采用蛋白免疫印迹(Western blotting)技术检测大鼠角膜微管相关蛋白轻链3(microtubule-associatedprotein 1 light chain 3,LC3)、P62和Beclin-1的表达,应用实时聚合酶链式反应(real-time polymerase chain reaction,RT-PCR)技术检测角膜LC3 mRNA的表达。结果与正常组比较,模型组大鼠泪液分泌量降低,泪膜破裂时间缩短,角膜荧光素染色评分升高,差异均具有统计学意义(P<0.05);角膜上皮细胞肿胀,线粒体明显损伤;角膜LC3Ⅱ和Beclin-1蛋白表达减少,P62蛋白表达增多,LC3 mRNA表达减少,差异均具有统计学意义(P<0.05)。与模型组比较,电针组大鼠泪液分泌量升高,泪膜破裂时间延长,角膜荧光素染色评分降低,差异均具有统计学意义(P<0.05);角膜上皮细胞轻微肿胀,有自噬小体形成;角膜LC3Ⅱ和Beclin-1蛋白表达增多,P62蛋白表达降低,LC3 mRNA表达增多,差异均具有统计学意义(P<0.05)。与模型组比较,假针组细胞器肿胀明显,线粒体损伤明显;其余指标差异均无统计学意义(P>0.05)。结论电针可以促进干眼大鼠角膜组织中自噬的发生,促进自噬可能是电针治疗干眼的潜在机制。
Objective To observe the effect of electroacupuncture in regulating autophagy in the cornea of dry eye rats.Method Male Sprague-Dawley(SD)rats were randomly divided into a normal group,a model group,an electroacupuncture group,and a sham acupuncture group.Except for the normal group,the other three groups received subcutaneous injection of Scopolamine solution plus exposure to air drought to establish dry eye rat models.After modeling,the electroacupuncture and sham acupuncture groups were given electroacupuncture or sham acupuncture,respectively.Before and after the intervention,tear film break-up time(BUT)and tear productionwere measured,and corneal fluorescein staining(CFS)was scored;rats’corneal ultrastructure was observed using transmission electron microscopy;Western blotting was adopted to determine the expression of microtubule-associated protein 1 light chain(LC3),P62,and Beclin-1 in the cornea;the real-time polymerase chain reaction(RT-PCR)technique was used to detect the expression of LC3 mRNA in rats’cornea.Result Compared with the normal group,the model group had a decreased tear production amount,shortened tear film BUT,and a higher CFS score,all showing statistical significance(P<0.05);the model group also presented swollen corneal epithelial cells with significant damage to mitochondria,reduced protein expression of corneal LC3Ⅱand Beclin-1,increased expression of P62 protein,and decreased expression of LC3 mRNA,and the differences were all statistically significant(P<0.05).Compared with the model group,the electroacupuncture group had an increased tear secretion amount,extended tear film BUT,and a lower CFS score,all showing statistical significance(P<0.05);in the electroacupuncture group,slightly swollen corneal epithelial cells were found,together with autophagosome formation,and the protein expression of corneal LC3Ⅱand Beclin-1 increased,the P62 protein expression dropped,and the LC3 mRNA expression increased,all showing statistical significance(P<0.05).Compared with the model group,t
作者
杨延婷
莫晓芬
马睿琦
杨光
师雅君
赵越
纪海洋
张丹
郭浩东
刘婕
马晓芃
YANG Yanting;MO Xiaofeng;MA Ruiqi;YANG Guang;SHI Yajun;ZHAO Yue;JI Haiyang;ZHANG Dan;GUO Haodong;LIU Jie;MA Xiaopeng(Shanghai Research Institute of Acupuncture and Meridian,Shanghai 200030,China;Eye&ENT Hospital of Fudan University,Shanghai 200030,China;Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200437,China)
出处
《上海针灸杂志》
CSCD
2024年第6期675-683,共9页
Shanghai Journal of Acupuncture and Moxibustion
基金
国家自然科学基金项目(81904302)
上海市自然科学基金项目(22ZR1458500,21ZR1460100)
上海市卫生健康委员会项目(20214Y0116,20234Y0066)。
关键词
针刺疗法
电针
干眼
角膜
自噬
大鼠
Acupuncture therapy
Electroacupuncture
Dry eye
Cornea
Autophagy
Rats
