养阴润目丸对苯扎氯铵诱导的干眼兔模型结膜上皮细胞焦亡的影响  

Effects of Yangyin Runmu Pill on pyroptosis of conjunctival epithelial cell induced by benzalkonium chloride in a rabbit dry-eye model

作　　者：李玲 李点[2] 张婷[2] 姚蕾 李炎 LI Ling;LI Dian;ZHANG Ting;YAO Lei;LI Yan(College of Chinese Medicine,Hunan University of Chinese Medicine,Changsha,Hunan 410208,China;The First Hospital of Hunan University of Chinese medicine,Changsha,Hunan 410007,China)

机构地区：[1]湖南中医药大学中医学院,湖南长沙410208 [2]湖南中医药大学第一附属医院,湖南长沙410007

出　　处：《湖南中医药大学学报》2023年第9期1570-1575,共6页Journal of Hunan University of Chinese Medicine

基　　金：国家自然科学基金面上项目(81674030);教育部产学合作协同育人项目(22100443904328)。

摘　　要：目的研究养阴润目丸对干眼兔结膜上皮细胞焦亡的调控作用。方法将40只雌雄各半的健康普通级新西兰兔采用随机数字表法,取8只作为空白组,剩余32只均采用双眼局部滴注0.1%苯扎氯铵溶液制备干眼模型,模型成功后随机分为空白组,模型组,养阴润目丸低、中、高剂量组,每组8只。养阴润目丸低、中、高剂量组分别按剂量0.435、0.869、1.738 g·(kg·d)^(-1)给予养阴润目丸水溶液灌胃。连续灌胃2周后,分别进行泪液分泌试验(Schirmer's test,SIT)、泪膜破裂时间(break up time,BUT)和角膜荧光素染色评分;取兔结膜上皮细胞,透射电镜下观察结膜上皮的细胞形态;Western blot检测结膜中NOD样受体家族核苷酸结合寡聚化结构域样受体蛋白3(NOD-like receptor family pyrin domain-containing protein 3,NLRP3)、半胱氨酸天冬氨酸特异性蛋白酶-1(cysteine aspartate-specific proteinase-1,Caspase-1)、消化道皮肤素D(Gasdermin D,GSDMD)蛋白表达;ELISA法检测兔血清中炎症因子白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-18(interleukin-18,IL-18)含量。结果与空白组相比,模型组实验兔SIT明显减少、泪膜BUT明显缩短、角膜荧光素染色评分明显升高(P<0.01,P<0.05);兔结膜上皮细胞超微结构可见明显肿胀、变形、破裂、皱缩等细胞损伤;兔结膜上皮细胞中NLRP3、Caspase-1、GSDMD的蛋白表达明显升高(P<0.05);兔血清中IL-1β、IL-18含量明显增多(P<0.05)。与模型组相比,养阴润目丸各剂量组SIT明显增多、泪膜BUT明显延长、角膜荧光素染色评分明显降低(P<0.05);兔结膜上皮细胞中NLRP3、Caspase-1、GSDMD的蛋白表达明显降低(P<0.05);兔血清中IL-1β、IL-18含量明显减少(P<0.05)。结论养阴润目丸可以通过降低干眼兔结膜上皮细胞中NLRP3、Caspase-1、GSDMD蛋白表达,抑制结膜上皮细胞发生焦亡,减少炎症细胞因子IL-1β、IL-18分泌,从而阻断相关级联反应,起到增加泪液分�Objective To study the effects of Yangyin Runmu Pill(YYRMP)on pyroptosis of conjunctival epithelial cells in dry-eye rabbits.Methods Eight rabbits were randomly selected among 20 male and 20 female New Zealand rabbits as the blank group.The remaining 32 rabbits were used to prepare the dry-eye models by local infusion of 0.1%benzalkonium chloride solution into both eyes.After successful modeling,they were randomly subdivided into blank group,model group,and low-,medium-,and high-dose YYRMP groups,with 8 rabbits in each group.The low-,medium-,and high-dose YYRMP groups were treated with YYRMP solution by gavage at dosages of 0.435,0.869,and 1.738 g·(kg·d)^(-1),respectively.After continuous gavage for 2 weeks,Schirmer's test(SIT),break up time(BUT),and corneal fluorescence staining score were performed respectively;rabbit conjunctival epithelial cells were observed by transmission electron microscopy.The expressions of NOD-like receptor family pyrin domain-containing protein 3(NLRP3),cysteine aspartate-specific proteinase-1(Caspase-1),and Gasdermin D(GSDMD)protein in the conjunctiva were checked by Western blot;the content of serum inflammatory factors interleukin-1β(IL-1β)and interleukin-18(IL-18)in rabbit serum was determined by ELISA.Results Compared with the blank group,the experimental rabbits in the model group showed significantly lower SIT,shorter tear film BUT,and higher fluorescence staining scores(P<0.01,P<0.05);the ultrastructure of rabbit conjunctival epithelial cells showed obvious swelling,deformation,rupture,shrinkage,and other cellular damage;the protein expressions of NLRP3,Caspase-1,and GSDMD and the content of IL-1β,IL-18 increased(P<0.05).Compared with the model group,the experimental rabbits in YYRMP groups showed higher SIT,longer tear film BUT,and lower fluorescence staining scores(P<0.05);the protein expressions of NLRP3,Caspase-1,and GSDMD in rabbit conjunctival epithelial cells were reduced(P<0.05);the content of IL-1βand IL-18 decreased(P<0.05).Conclusion YYRMP can reduce the expre

关 键 词：干眼 养阴润目丸 苯扎氯铵 结膜上皮细胞 细胞焦亡 

分 类 号：R276.7[医药卫生—中医五官科学]