川陈皮素通过激活囊性纤维化跨膜传导调节因子(CFTR)治疗小鼠2型糖尿病干眼的实验研究  

Experimental study on the effect of nobiletin in the treatment of dry eyes in mice with type 2 diabetes mellitus by activating cystic fibrosis transmembrane conductance regulators

作　　者：王珏 张婕 王卓欣 张鑫 苏丽萍 WANG Jue;ZHANG Jie;WANG Zhuoxin;ZHANG Xin;SU Liping(Department of Ophthalmology,the Second Affiliated Hospital of Air Force Medical University,Xi’an 710038,Shaanxi Province,China;Military Personnel Medical Care Center,the Second Affiliated Hospital of Air Force Medical University,Xi’an 710038,Shaanxi Province,China;Department of Ophthalmology,Xi’an Aier Ancient City Eye Hospital,Xi’an 710014,Shaanxi Province,China)

机构地区：[1]空军军医大学第二附属医院眼科,陕西省西安市710038 [2]空军军医大学第二附属医院军队人员医疗保健中心,陕西省西安市710038 [3]西安爱尔古城眼科医院眼科,陕西省西安市710014

出　　处：《眼科新进展》2025年第1期15-21,共7页Recent Advances in Ophthalmology

基　　金：陕西省科技攻关重点研发计划项目(编号:2021SF-158);唐都医院科技创新发展基金资助项目(编号:2019LCYJ014)。

摘　　要：目的探究川陈皮素(NOB)治疗2型糖尿病干眼(T2DM-DE)的潜在价值及其对囊性纤维化跨膜传导调节因子(CFTR)表达的影响。方法通过高脂饮食联合链脲佐菌素建立T2DM小鼠模型,然后采用苯扎氯铵诱导T2DM-DE小鼠模型。将小鼠分为6组:对照组(正常小鼠)、T2DM-DE组、L-NOB组、M-NOB组、H-NOB组和CFTR抑制剂组,后5组建立T2DM-DE模型。对照组和T2DM-DE组小鼠灌胃含体积分数0.5%吐温80的PBS,L、M、H-NOB组小鼠分别灌胃剂量为50、100、200 mg·kg^(-1)·d^(-1)的NOB溶液,CFTR抑制剂组小鼠灌胃200 mg·kg^(-1)·d^(-1)的NOB溶液的同时腹腔注射1 mg·kg^(-1)·d^(-1)的CFTR(inh)-172,均给药4周。分别检测小鼠空腹血糖(FPG)、泪液分泌量、泪膜破裂时间(BUT)、角膜荧光素钠染色分级、泪液中白细胞介素(IL)-1β、IL-6、IL-8和肿瘤坏死因子-α(TNF-α)水平。分别进行小鼠结膜杯状细胞PAS染色和角膜TUNEL染色。采用RT-qPCR和Western blot检测小鼠角膜CFTR、Bax和Bcl-2的mRNA和蛋白表达水平。结果与对照组比较,T2DM-DE组小鼠的FPG升高,泪液中IL-1β、IL-6、IL-8和TNF-α水平升高,角膜荧光素钠染色分级和TUNEL染色阳性细胞率均升高(均为P<0.05);泪液分泌量、BUT和结膜杯状细胞数量均降低,CFTR的mRNA和蛋白相对水平均降低(均为P<0.05)。与T2DM-DE组比较,L-NOB组、M-NOB组和H-NOB组小鼠的FPG降低,泪液中IL-1β、IL-6、IL-8和TNF-α水平降低,角膜荧光素钠染色分级和TUNEL染色阳性细胞率均降低(均为P<0.05);泪液分泌量、BUT和结膜杯状细胞数量均升高,CFTR的mRNA和蛋白相对水平均升高(均为P<0.05)。与H-NOB组比较,CFTR抑制剂组的FPG升高,泪液中IL-1β、IL-6、IL-8和TNF-α水平升高,角膜荧光素钠染色分级和TUNEL染色阳性细胞率均升高(均为P<0.05);泪液分泌量、BUT和结膜杯状细胞数量均降低,CFTR的mRNA和蛋白相对水平均降低(均为P<0.05)。结论NOB通过激活CFTR降低T2DM-DE小鼠血糖水平,�Objective To investigate the potential value of nobiletin(NOB)in the treatment of dry eyes in type 2 diabetes mellitus(T2DM-DE)and its effect on the expression of cystic fibrosis transmembrane conductance regulators(CFTRs).Methods The T2DM mouse model was established by a high-fat diet combined with streptozotocin,and then the T2DM-DE mouse model was induced by benzalkonium chloride.These mice were divided into 6 groups:the NC group,the T2DM-DE group,the L-NOB group,the M-NOB group,the H-NOB group,and the CFTR inhibitor group.Mice in the NC and T2DM-DE groups were provided with phosphate buffer saline(PBS)containing 0.5%Tween80 through oral gavage;those in the L-NOB,M-NOB,and H-NOB groups were provided with 50,100,and 200 mg·kg^(-1)·d^(-1) NOB solutions,respectively,through oral gavage;those in the CFTR inhibitor group were provided with 200 mg·kg^(-1)·d^(-1) NOB solution through oral gavage and intraperitoneally injected with 1 mg·kg^(-1)·d^(-1) CFTR(inh)-172.The intervention in these groups was provided for 4 weeks.The fasting blood glucose(FPG),tear secretion,tear film break-up time(BUT),corneal fluorescein sodium staining grade,and interleukin-1β(IL-1β),IL-6,IL-8,and tumor necrosis factor-α(TNF-α)levels in tears of mice were detected.Besides,the periodic acid-Schiff(PAS)staining of conjunctival goblet cells and the terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)staining of the cornea were performed respectively.The mRNA and protein expression levels of CFTRs,Bax,and Bcl-2 in the cornea were measured by the real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot.Results Compared with the NC group,the FPG of mice in the T2DM-DE group increased;the level of IL-1β,IL-6,IL-8,and TNF-αin tears increased;the corneal fluorescein sodium staining grade and TUNEL positive rate increased(all P<0.05);the tear secretion,BUT,and the number of conjunctival goblet cells decreased;the mRNA and protein expression levels of CFTRs decreased(all P<0.05).Compared with the T2DM-DE gr

关 键 词：2型糖尿病 干眼 川陈皮素 囊性纤维化跨膜传导调节因子 血糖 炎症 结膜杯状细胞 角膜细胞 

分 类 号：R777[医药卫生—眼科]