电针通过调控TLR4介导的炎症信号通路改善糖尿病干眼大鼠角膜炎症的机制研究  

Mechanism of electroacupuncture on improving corneal inflammation in diabetic dry eye rats by regulating Toll-like receptor 4-mediated inflammatory signaling pathway

作　　者：傅张倚天 万咪咪 赖思华 孙心怡 李小颖 高卫萍 FU Zhangyitian;WAN Mimi;LAI Sihua;SUN Xinyi;LI Xiaoying;GAO Weiping(Department of Ophthalmology,Affiliated Hospital of Nanjing University of Traditional Chinese Medicine,Nanjing 210024,Jiangsu Province,China;Department of Acupuncture and Rehabilitation,Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210024,Jiangsu Province,China;Department of Endocrinology,Affiliated Hospital of Nanjing University of Traditional Chinese Medicine,Nanjing 210024,Jiangsu Province,China)

机构地区：[1]南京中医药大学附属医院眼科,江苏省南京市210024 [2]南京中医药大学附属医院针灸康复科,江苏省南京市210024 [3]南京中医药大学附属医院内分泌科,江苏省南京市210024

出　　处：《眼科新进展》2024年第1期24-29,共6页Recent Advances in Ophthalmology

基　　金：国家自然科学基金(编号:82074526)。

摘　　要：目的观察电针对糖尿病干眼大鼠角膜Toll样受体4(TLR4)介导的炎症信号通路的影响,探讨电针治疗糖尿病干眼的作用机制。方法健康雄性SD大鼠32只采用高糖高脂饲料喂养4周后,腹腔注射链脲佐菌素(30 mg·kg^(-1))12周建立2型糖尿病大鼠模型。将造模成功的25只糖尿病干眼大鼠随机分为模型组(不做干预)、电针组(选取“睛明”“攒竹”“丝竹空”“太阳”“瞳子髎”针刺,电针接“攒竹”“瞳子髎”,每次15 min,每天1次)、假针刺组(穴位刺激处同电针组,但用钝头针点刺治疗,不刺入)、氟米龙组(双眼点滴1 g·L^(-1)氟米龙滴眼液,分别在每天8点钟、13点钟、18点钟进行干预,每天3次,每次1滴),每组6只,共干预2周。另选取6只正常雄性SD大鼠作为空白组。检测各组大鼠造模前、造模后及干预后的随机血糖、泪膜破裂时间(BUT)、泪液分泌量、角膜荧光素染色(FL)评分及角膜机械知觉阈值(CTT);HE染色观察各组大鼠角膜形态变化;免疫荧光组织化学染色法检测各组大鼠角膜TLR4阳性表达;Western blot检测角膜中TLR4、磷酸化核因子-κB P65(P-NF-κB P65)、白细胞介素(IL)-1β及IL-18表达水平。结果造模后,与空白组比较,各实验组大鼠BUT、泪液分泌量、CTT均下降,FL均升高,差异均有统计学意义(均为P<0.01);干预后,与模型组比较,电针组大鼠FL降低,BUT、泪液分泌量及CTT均升高,差异均有统计学意义(均为P<0.05)。角膜HE染色显示,干预后,模型组和假针刺组大鼠角膜表面不光滑,角膜上皮细胞增厚且排列紊乱;电针组及氟米龙组大鼠角膜表面光滑,角膜上皮细胞排列整齐。干预后,与空白组比较,其他各组大鼠角膜TLR4表达均升高,差异均有统计学意义(均为P<0.05);与模型组比较,电针组及氟米龙组大鼠角膜TLR4表达均降低,差异均有统计学意义(均为P<0.01)。干预后,与空白组比较,模型组与假针刺组大鼠角膜TLR4、P-NObjective To observe the effect of electroacupuncture on corneal Toll-like receptor 4(TLR4)-mediated inflammatory signaling pathway in diabetic dry eye rats and to explore the mechanism of electroacupuncture in the treatment of diabetic dry eye.Methods A type 2 diabetic rat model was established in 32 healthy male Sprague-Dawley(SD)rats by intraperitoneal injection of streptozotocin(30 mg·kg^(-1))for 12 weeks after feeding with high-sugar and high-fat diet for 4 weeks.Twenty-five successfully modeled diabetic dry eye rats were randomly divided into the model group(non-intervention),electroacupuncture group(the“Jingming”,“Cuanzhu”,“Sizhukong”,“Taiyang”and“Tongziliao”acupoints were treated with acupuncture,and then“Cuanzhu”and“Tongzilao”acupoints were treated with electroacupuncture,15 min for each time,once a day),sham acupuncture group(blunt-tip needle pricking was performed at the same acupoints as the electroacupuncture group),and fluorometholone group(1 g·L^(-1)fluorometholone eye drops were used in both eyes at 8 o’clock,13 o’clock,and 18 o’clock,1 drop each time),with 6 rats in each group,lasting for 2 weeks.Another 6 healthy male SD rats were selected as a blank group.Random blood glucose,tear film breakup time(BUT),tear secretion,corneal fluorescein staining(FL)score,and corneal touch threshold(CTT)of rats in each group were detected before modeling,after modeling,and after the intervention.Hematoxylin and Eosin(HE)staining was performed to observe the corneal morphologic changes in each group.Immunofluorescence histochemical staining was adopted to detect the corneal TLR4-positive expression in each group.The TLR4,phosphorylated nuclear factor-kappa P65(P-NF-κB P65),interleukin(IL)-1β,and IL-18 expression levels in the cornea were detected by Western blot.Results After modeling,compared with the blank group,BUT,tear secretion and CTT decreased and FL increased in all experimental groups,and the differences were statistically significant(all P<0.01).After the intervent

关 键 词：干眼 2型糖尿病 炎症 电针 Toll样受体4 核因子ΚB 

分 类 号：R777.34[医药卫生—眼科]