摘要
目的基于蛋白激酶A(PKA)/环磷酸腺苷反应元件结合蛋白(CREB)信号通路,探讨虎杖苷(PD)对干眼症(DED)大鼠炎症反应的影响及潜在机制。方法将雄性SD大鼠随机分为空白对照组(Normal组)、模型组(单纯DED组)、0.05%PD组、0.5%PD组和0.5%PD+PKA抑制剂H-89组(0.5%PD+H-89组),每组15只。除Normal组外,其余各组大鼠通过眼球表面注射氢溴酸东莨菪碱12.5 mg/d的方式制备DED模型;与此同时,各药物组给予相应药液(0.5%、0.05%的PD,滴眼,每天3次;1 mg/kg的H-89,腹腔注射,每天1次),共7 d。检测各组大鼠的泪液分泌量、角膜荧光素染色评分和结膜组织杯状细胞密度,观察其角膜组织病理变化,检测其角膜组织中炎症因子(肿瘤坏死因子α、白细胞介素1β、白细胞介素6)水平和PKA/CREB信号通路相关蛋白的表达情况。结果与Normal组比较,单纯DED组大鼠角膜组织上皮层增厚,基质层细胞排列紊乱、疏松且部分缺失,核间距较大,有大量炎症细胞浸润;泪液分泌量、结膜组织杯状细胞密度和PKA、CREB蛋白的磷酸化水平均显著降低,角膜荧光素染色评分和角膜组织中炎症因子水平均显著升高(P<0.05)。与单纯DED组比较,0.05%PD、0.5%PD组大鼠的角膜组织病理损伤有所缓解,各定量指标均显著改善,且0.5%PD组的改善效果更优(P<0.05);而H-89可显著逆转PD对各指标的改善作用(P<0.05)。结论PD可增加DED大鼠的泪液分泌量和结膜组织杯状细胞密度,减轻其角膜组织炎症反应和病理损伤,上述作用与激活PKA/CREB信号通路有关。
OBJECTIVE To investigate the effects of polydatin(PD)on the inflammatory response in dry eye disease(DED)rats and its potential mechanism based on protein kinase A(PKA)/cyclic adenosine monophosphate response element binding protein(CREB)signaling pathway.METHODS Male SD rats were randomly divided into blank control group(normal group),model group(simple DED group),0.05%PD group,0.5%PD group and 0.5%PD+PKA inhibitor H-89 group(0.5%PD+H-89group),with 15 rats in each group.Except for the normal group,the rats in other groups were prepared with a DED model by injecting scopolamine hydrobromide 12.5 mg/d onto the surface of eyeball.At the same time,each drug group was given corresponding liquid medicine(0.5%or 0.05%PD,eye drip,3 times a day;1 mg/kg H-89,intraperitoneal injection,once a day)for 7 d in total.The tear secretion,corneal fluorescein staining score and conjunctival goblet cell density of rats were detected in each group;the pathological changes of corneal tissues were observed,and the levels of inflammatory factors(tumor necrosis factor-α,interleukin-1β,interleukin-6)and the expression of PKA/CREB signaling pathway-related proteins were detected in corneal tissues.RESULTS Compared with normal group,the corneal epithelium of rats in the simple DED group was thickened,stromal layer cells were disordered and partially absent,nuclear spacing was larger,and a large number of inflammatory cells infiltrated;the tear secretion,conjunctival goblet cell density and the phosphorylation levels of PKA and CREB in corneal tissues were reduced significantly,while the corneal fluorescein staining score and the levels of inflammatory factors in corneal tissue were increased significantly(P<0.05).Compared with the simple DED group,the pathological injuries of corneal tissues of rats in 0.05%PD and 0.5%PD groups were alleviated,each quantitative index was significantly improved,and the improvement effect of 0.5%PD group was more obvious(P<0.05).H-89 could reverse the improvement effect of PD on each index significantly(P<0.05).CONCLUSIONS PD can increase tear secretion and the conjunctival goblet cell density,and reduce the inflammatory response and pathological injury of corneal tissue in DED rats.The above effects are related to the activation of PKA/CREB signaling pathway.
作者
赵丽琼
魏瑞华
ZHAO Liqiong;WEI Ruihua(Optometry Center,Tianjin Medical University Eye Hospital/School of Optometry/Eye Institute/Tianjin Branch of National Clinical Research Center for Eye,Ear,Nose and Throat Diseases/Tianjin Key Laboratory of Retinal Functions and Diseases,Tianjin 300384,China;Dept.of Refractive Keratology,Tianjin Medical University Eye Hospital/School of Optometry/Eye Institute/Tianjin Branch of National Clinical Research Center for Eye,Ear,Nose and Throat Diseases/Tianjin Key Laboratory of Retinal Functions and Diseases,Tianjin 300384,China)
出处
《中国药房》
CAS
北大核心
2024年第18期2213-2218,共6页
China Pharmacy
基金
国家自然科学基金项目(No.82070929)
天津市医学重点学科(专科)建设项目(No.TJYXZDXK-037A)。
作者简介
第一作者:赵丽琼,医师,硕士。研究方向:眼表疾病及眼视光学。E-mail:zhaoliqiong001@126.com;通信作者:魏瑞华,主任医师,博士。研究方向:眼表疾病及眼视光学。E-mail:rwei@tmu.edu.cn。
