鬼针草水提物对雄激素缺乏性干眼大鼠泪液分泌和泪腺炎症的影响

Effects of aqueous extract of Bidens pilosa L. on tear secretion and lacrimal gland inflammation of dry eye due to androgen deficiency in rats

目的研究鬼针草水提物对雄激素缺乏性干眼大鼠泪液分泌和泪腺炎症的影响,并探讨其作用机制。方法 24只雌性Wistar大鼠随机分为4组,每组6只。正常对照组无任何处理;生理盐水组予生理盐水灌胃,非那雄胺组予非那雄胺灌胃,鬼针草治疗组予非那雄胺和鬼针草提取物灌胃,均连续4周。实验前、实验后7 d、14 d、21 d及28 d进行泪液分泌量、泪膜破裂时间和角膜荧光素染色检测。实验后28 d后处死实验动物,采用细胞因子抗体芯片技术及Western blot分析泪腺和角膜炎症因子变化,并对泪腺进行组织病理观察。结果非那雄胺给药14 d后可有效诱导大鼠干眼,与正常对照组比较,非那雄胺组大鼠的泪液分泌量明显减少,泪膜破裂时间明显缩短,角膜荧光素染色评分明显增加,泪腺组织可见大量的炎症细胞浸润。鬼针草治疗组大鼠的泪液分泌量、泪膜破裂时间及角膜荧光素染色情况均明显好于非那雄胺组,其泪腺结构较完整,未见明显的炎症细胞浸润。细胞因子抗体芯片和Western blot结果显示,非那雄胺组的促炎症因子白细胞介素-1β(IL-1β)、IL-6和肿瘤坏死因子-α(TNF-α)以及抗炎细胞因子IL-4、IL-10的表达水平均高于正常对照组。与非那雄胺组相比,鬼针草治疗组的IL-1β,Fas配体(Fas L)及TNF-α表达水平明显降低。结论鬼针草水提物能有效改善雄激素缺乏性干眼大鼠的泪液分泌,稳定泪膜,并能抑制泪腺组织炎症反应。

OBJECTIVE To investigate the effect of the aqueous extract of Bidens pilosa L.（Bp） on tear secretion and lacrimal gland inflammation of dry eye due to androgen deficiency,and further to discuss its possible mechanisms.METHODS Twenty-four Wistar rats were randomly divided into four groups：Group C（control）,Group S（physiological saline）,Group F（oral finasteride）,and Group B（finasteride and Bp by oral）.Tear volume was mea-sured by phenol red-impregnated cotton threads with anesthesia.Tear film breakup time（BUT） and corneal epithelial damage were evaluated by fluorescein staining.Animals were sacrificed at the 28th day,gene microarray analysis for inflammatory cytokines of lacrimal gland and cornea,Western blot were also performed.Lacrimal gland was evaluat-ed histopathologically by light microscopy.RESULTS Fourteen days of Finasteride administration effectively induced dry eye in rats.Rats in group F had significantly higher fluorescein staining scores and lower tear volume and BUT than counterparts in group C,and notable inflammatory cell infiltration were observed in the lacrimal gland of rats in group F.The fluorescein staining score,tear volume and BUT significantly improved with Bp treatment in the group B,and the structures of the lacrimal gland were well maintained without significant lymphocyte infiltration.Cytokine antibody array data and Western blot showed that,expression levels of pro-inflammatory cytokines including IL-1β,IL-6 and TNF-α as well as anti-inflammatory cytokines,such as IL-4 and IL-10 in group F were higher than result of group C.Levels of IL-1β,Fas L and TNF-α in group B were lower than result in group F.CONCLUSIONS The extract of Bp appeared to be effective on dry eye owing to decreased androgen in that it promoted the tear secretion,stabled tear film and inhibited the inflammation of lacrimal gland.