疏肝养阴法调控干眼局部微环境的分子机制研究

Dry eye is a common, frequently-occurring disease and has gradually become a hot topic of ophthalmic research. The main pathogenesis of dry eye is non-infectious immune inflammation. One of the main treatments is artificial tear drops which is not cause treatment; second is hormones and immunosuppressive agents drops which need long treatment time and lead to the inevitable side effects, such as glaucoma, cataracts, eye discomfort and ect. Although dry eye is an ocular disease, but due to systemic pathological changes. The "holism" and "Syndrome Differentiation treatment" is the essence of traditional Chinese medicine. Chinese medicine believes that "the liver having its specific body opening in the eye "," Tears being an expression of of liver fluid". When liver stagnation and yin deficiency, the body fluid cannot spread upward, so eyes become drying. As a result, we use "soothe the liver and nourishing yin" to treat dry eye, pre-clinical studies confirm that this method can control symptoms and prolong the tear film breakup time, and promote tear secretion. The main possible mechanism of these may be immune suppression. In order to investigate the mechanism further, we establish this subject. In this study, we will observe the soothe the liver and nourishing yin medicine lead to the expression changes of inflammatory cytokines in the conjunctival goblet cells and corneal epithelial cells; we will also establish two dry eye animal model to observe the role of soothe the liver and nourishing yin medicine, keratoconjunctivitis cytokine changes due to it and its possible effect like glucocorticoid.

干眼是眼科临床常见病、多发病，近几年逐渐成为眼科研究的热点。干眼的主要发病机理是非感染性免疫炎症。目前的主要治疗方法之一是人工泪液滴眼，但不能针对病因治疗；之二是激素及免疫抑制剂滴眼，但治疗时间长，有导致青光眼、白内障、眼部不适等不可避免的副作用。干眼虽是眼局部病变，但发生原因为全身病理变化，"整体观念"和"辨证论治"是中医认识和治疗疾病的精髓。中医认为"肝开窍于目"，"泪为肝液"，肝郁阴虚，津液不能上承，故发干眼。由此，我们以疏肝养阴法治疗干眼，前期的临床研究证实该法能控制症状，延长泪膜破裂时间，促进泪液分泌，考虑主要可能的作用机理是免疫抑制，为进一步探讨其作用机制，立此课题。本研究采用疏肝养阴中药，观察其对结膜杯状细胞和角膜上皮细胞炎症因子表达的影响；并建立两种干眼动物模型，观察疏肝养阴中药的治疗作用，及对角结膜炎性细胞因子改变及可能的拟糖皮质激素作用。

项目的背景.干眼是全身病理变化所导致的局部病变，主要病因病机有免疫炎性因素、性激素失衡、神经机能障碍及细胞凋亡等导致泪液质和量，及泪膜稳定性下降。目前国际上对干眼的治疗，主要为人工泪液点眼，此种方法仅为对症治疗而非对因治疗。而大多数患者使用人工泪液后并不能缓解干涩症状，甚而至于症状加重。因此，本课题组认为 “肝郁阴虚”为本病的主要病机，由此提出 “疏肝养阴”为其主要对因治疗原则，进而研究疏肝养阴法调控干眼局部免疫炎性微环境的作用机制。.主要研究内容.分为动物体内实验及体外细胞实验，①动物实验采用两种国际公认的动物模型：分别为BAC点眼和东莨菪碱皮下注射/吹风进行干眼造模，行干眼各项指标（BUT、Schimmer 试验、角膜荧光染色）观察均证实造模成功。检测干眼模型小鼠及经低、中、高剂量组逍生散颗粒剂治疗后小鼠泪液及角膜组织中IL-1、IL-6、TNF-α、TGF、TLR等炎性因子的表达及杯状细胞数量，从而得出结论：逍生散颗粒剂可增加泪液中TGF-β的含量，降低IL-1、IL-6、TLR、TNF-α、TLR等炎性因子的表达，逍生散颗粒剂可能具有抑制炎性细胞因子的生成，减少促炎细胞因子的释放，并防止杯状细胞之凋亡而发挥治疗作用。两种干眼实验动物模型检测结果相吻合。② 进行体外细胞的培养及鉴定，制备了干眼角膜上皮细胞损伤模型：课题组率先制备角膜上皮干眼损伤模型，进而研究不同剂量组的逍生散颗粒剂对干眼角膜上皮细胞损伤模型的干预及调节作用。.重要结果.课题组认为疏肝养阴法治疗干眼是通过调控干眼局部免疫炎性细胞因子水平及改善性激素水平，减少促炎细胞因子释放，并防止杯状细胞凋亡发挥其药理作用。.科学意义.1、探讨中医中药对干眼的对因治疗。2、从整体观念出发，调整全身阴阳平衡，达到对局部病变部位治疗的目的。3、结合本病的发病机理，研究疏肝养阴法对机体免疫炎性因子的调节作用。4、为开发有效治疗干眼口服中成药物填补空白。5、没有眼药水局部点眼所带来的一系列副作用（如眼药水中的防腐剂对眼表的损害）。