摘要
目的 从调节大鼠眼表功能及结膜组织中NOD样受体热蛋白结构域 3(NOD-like receptor pyrin domain containing 3,NLRP3)炎症小体及相关蛋白角度探讨电针对干眼(Dry eye,DE)大鼠的作用.方法 将32 只雄性SD大鼠随机分为正常组、模型组、电针组和假电针组 4 组,每组各8 只.采用氢溴酸东莨菪碱皮下注射结合吹风法制备DE模型.造模成功后电针组取双侧"丝竹空""风池"穴进行电针干预,每日1 次,每次20 min.假电针组取穴同电针组,穴位连接电针但不通电.两组各干预 7d.检测各组干预前后的泪液分泌量、泪膜破裂时间;观察结膜组织病理学变化;采用Western blot和免疫组化法检测结膜组织中NLRP3、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a CARD,ASC)、半胱天冬酶(Caspase)-1、白细胞介素(interleukin,IL)-1β、IL-18 蛋白表达的变化.结果 与正常组比较,模型组泪液分泌量和泪膜破裂时间显著降低(P<0.05),结膜组织中NLRP3、ASC、Caspase-1、IL-1β、IL-18 蛋白表达均升高(P<0.05);与模型组比较,电针组泪液分泌量和泪膜破裂时间均显著升高(P<0.05),结膜组织中NLRP3、ASC、Caspase-1、IL-1β、IL-18 蛋白表达均降低(均P<0.01);且电针组IL-1β和IL-18 蛋白表达较假电针组均显著降低(P<0.05).结论 电针能改善DE大鼠的眼表功能,下调结膜NLRP3 炎症小体及下游炎性因子IL-1β、IL-18 的表达.电针可能通过抑制NLRP3 炎症小体信号通路减轻DE大鼠眼表炎症.
Abstract
Objective Exploring the role of electroacupuncture in dry eye(DE)rats in terms of modulation of ocular surface function and NLRP3 inflammasome and related proteins in conjunctival tissue.Methods Thirty-two male SD rats were randomly divided into four groups:the normal group,the model group,the electroacupuncture group,and the sham electroacupuncture group,with eight rats in each group.The DE model was developed by subcutaneous injection of scopolamine hydrobromide with exposure to an air draft in the latter four groups.After successful modeling,the electroacupuncture group took bilateral"Sizhukong"and"Fengchi"acupoints for electroacupuncture intervention once a day for 20 min each time.The sham electroacupuncture group took the same acupoints as the electroacupuncture group,and the acupoints were connected to the electroacupuncture needle but not electrified.The tear secretion and tear film breakup time of the rats were measured before and after the intervention for 7 days.The histopathological changes of conjunctiva tissues were observed.The changes in NLRP3,ASC,Caspase-1,IL-1β,and IL-18 protein expression in conjunctival tissue were detected by Western blot and immunohistochemistry.Results Compared with the normal group,tear secretion and tear film rupture time were significantly reduced in the model group(P<0.05),and NLRP3,ASC,Caspase-1,IL-1β,and IL-18 protein expression in conjunctival tissues were elevated(P<0.05);compared with the model group,tear secretion and tear film rupture time in the electroacupuncture group were significantly elevated(P<0.05),and the protein expression of NLRP3,ASC,Caspase-1,IL-1β,and IL-18 in conjunctival tissues were all reduced(all P<0.01).The expression of IL-1β and IL-18 proteins in the electroacupuncture group was significantly lower than that in the sham electroacupuncture group(P<0.05).Conclusion Electroacupuncture improved the ocular surface function of DE rats and down-regulated the expression of the conjunctival NLRP3 inflammasome and downstream inflammatory factors IL-1β and IL-18.Electroacupuncture may reduce ocular surface inflammation in DE rats by inhibiting the NLRP3 inflammasome signaling pathway.
基金项目
上海市自然科学基金项目(22ZR1458500)
上海市自然科学基金项目(21ZR1460100)
上海市卫生健康委员会项目(20214Y0116)
上海市卫生健康委员会项目(202040249)
上海市"科技创新行动计划"启明星项目(扬帆专项)(23YF1442000)
国家科技期刊平台
NSTL
万方数据