基于NOD样受体蛋白3/半胱天冬酶-1信号通路探究槲寄生滴眼液对干眼症模型小鼠的治疗效果  

Therapeutic effects of mistletoe eye drops on dry eye syndrome in mice via the NOD-like receptor protein 3/Caspase-1 signaling pathway

作　　者：吴一峰 姜蕾 唐丹燕 邓珍 WU Yifeng;JIANG Lei;TANG Danyan;DENG Zhen(Graduate School of Zhejiang Chinese Medicine University,Hangzhou,Zhejiang 310053,China;Department of Ophthalmology,the First People's Hospital of Linping District in Hangzhou City,Hangzhou,Zhejiang 311100,China)

机构地区：[1]浙江中医药大学研究生院,杭州310053 [2]杭州市临平区第一人民医院眼科,杭州311100

出　　处：《浙江中西医结合杂志》2024年第7期611-618,共8页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

摘　　要：目的基于NOD样受体蛋白3(NLRP3)/半胱天冬酶-1(Caspase-1)信号通路探究槲寄生滴眼液对干眼症模型小鼠的治疗效果。方法将干眼症模型小鼠(采用苯扎氯铵溶液滴眼造模)随机分为模型组(以生理盐水干预)、槲寄生滴眼液组(槲寄生滴眼液滴眼5μL,每天2次)、NLRP3/Caspase-1通路抑制剂(VX-765)组(腹腔注射50 mg/kg VX-765,每天1次)、槲寄生滴眼液+VX-765组(槲寄生滴眼液滴眼5μL,每天2次,腹腔注射50 mg/kg VX-765,每天1次),每组12只。另取12只健康小鼠作为对照组(以生理盐水干预)。各组均持续给药14 d。酚红棉线测定泪液分泌情况;苏木精-伊红染色观察结膜上皮和泪腺组织病理改变情况;酶联免疫吸附试验测定血清白细胞介素(IL)-18、IL-1β水平;荧光定量PCR和蛋白质印迹法分别测定结膜上皮和泪腺组织NLRP3、Caspase-1 mRNA和蛋白表达。结果与对照组比较,模型组结膜上皮细胞排列紊乱,杯状细胞数量减少,固有层可见淋巴细胞浸润等病理损伤,泪腺组织呈现腺泡空泡样改变,结构不清晰,大小不均一,各小叶及腺泡间细胞排列紊乱等病理变化,泪液分泌量(酚红棉线被浸湿的长度,下同)[(2.66±0.42)mm比(6.14±1.09)mm,P<0.05]显著减少,血清IL-18[(53.16±6.95)pg/mL比(15.04±2.14)pg/mL,P<0.05]、IL-1β[(57.28±7.11)pg/mL比(24.51±3.06)pg/mL,P<0.05]、结膜上皮及泪腺组织NLRP3、Caspase-1 mRNA和蛋白[NLRP3 mRNA:(3.05±0.43)比(1.00±0.05),P<0.05;Caspase-1mRNA:(4.41±0.62)比(1.00±0.06),P<0.05;NLRP3蛋白:(1.33±0.21)比(0.17±0.03),P<0.05;Caspase-1蛋白:(1.51±0.26)比(0.20±0.04),P<0.05]表达显著升高。与模型组比较,槲寄生滴眼液组、VX-765组和槲寄生滴眼液+VX-765组结膜上皮和泪腺组织上述病理损伤减轻,泪液分泌量[(4.15±0.53)mm、(4.11±0.55)mm、(5.38±0.62)mm比(2.66±0.42)mm,P<0.05]显著增加,血清IL-18[(36.03±5.71)pg/mL、(35.92±5.66)pg/mL、(20.07±3.38)pg/mL比(53.16±6.95)pg/mL,P<0.05]、IL-1β[(40.85±5.0Objective To investigate the therapeutic effects of mistletoe eye drops on dry eye syndrome in mice via the NOD-like receptor protein 3(NLRP3)/Caspase-1 signaling pathway.Methods Dry eye syndrome model was induced by benzalkonium chloride solution eye drops in mice,which were then randomly divided into four groups:the model(intervened with saline),the mistletoe eye drops(5μL mistletoe eye drops,twice daily),the NLRP3/Caspase-1 pathway inhibitor(VX-765)(50 mg/kg VX-765 by intraperitoneal injection,once daily),and mistletoe eye drops+VX-765(5μL mistletoe eye drops,twice a daily,and 50 mg/kg VX-765 by intraperitoneal injection,once daily)groups,with 12 mice in each group.An additional 12 healthy mice served as a control group(intervened with normal saline).All treatments were lasted for 14 days.The tear secretion was measured using the phenol red thread test.Histopathological changes of conjunctival epithelium and lacrimal gland tissues were observed using hematoxylin-eosin staining.Serum levels of interleukin(IL)-18 and IL-1βwere determined using enzymelinked immunosorbent assay.mRNA and protein expression levels of NLRP3 and Caspase-1 in conjunctival epithelium and lacrimal gland tissues were determined using quantitative PCR and Western blotting.Results Compared with the control group,the model group showed significant pathological damages,including disordered arrangement of conjunctival epithelial cells,decreased number of goblet cells,and lymphocyte infiltration in the lamina propria.The lacrimal gland tissue showed significant pathological changes,including acinar vacuolar-like changes with unclear structure and uneven size,and disordered arrangement of cells within lobules and acini.The volume of tear secretion(length of phenol red thread wetting)[(2.66±0.42)mm vs.(6.14±1.09)mm,P<0.05]was significantly reduced.Serum levels of IL-18[(53.16±6.95)pg/mL vs.(15.04±2.14)pg/mL,P<0.05]and IL-1β[(57.28±7.11)pg/mL vs.(24.51±3.06)pg/mL,P<0.05]were significantly increased.The mRNA levels of NLRP3[(3.05±0.43)vs

关 键 词：小鼠 干眼症 槲寄生滴眼液 NOD样受体蛋白3 半胱天冬酶-1 泪液分泌量 结膜上皮 泪腺 

分 类 号：R777.34[医药卫生—眼科] R-332[医药卫生—临床医学]