养阴润目丸对干眼模型大鼠结膜上皮细胞中ICAM-1、p38MAPK、p-p38MAPK表达的影响

李点1，王超群2，廖亮英1，胡平2，彭银艳2

（1.湖南中医药大学第一附属医院，湖南长沙410007；2.湖南中医药大学，湖南长沙410208）

〔摘要〕目的研究养阴润目丸对干眼模型大鼠结膜上皮细胞中细胞间粘附分子-1（ICAM-1）、丝裂原活化蛋白激酶p38（p38MAPK）、磷酸化-丝裂原活化蛋白激酶p38（p-p38MAPK）表达的影响。方法将60只SD雄性大鼠随机分为5组（正常组、假手术组、养阴润目丸组、新泪然组、模型组），每组12只。除正常组与假手术组外均采用去势法制造干眼模型。造模成功后一周开始分别给药治疗。用药3个月后，分别进行泪液分泌试验（SIT）及泪膜破裂时间（BUT）的检测，取大鼠结膜上皮细胞，采用免疫组化法检测p38MAPK的表达，采用Western blot法检测ICAM-1、p-p38MAPK的表达。结果与模型组比较，养阴润目丸组SIT明显增多，BUT明显延长；养阴润目丸可显著减少结膜上皮细胞中ICAM-1、p38MAPK、p-p38MAPK的表达，差异有统计学意义（P<0.01）。结论养阴润目丸可有效改善眼表症状，增加泪液分泌量，延长泪膜破裂时间，下调结膜上皮细胞中ICAM-1、p38MAPK、p-p38MAPK的表达。提示干眼的发病机制与炎症及p38MAPK信号通路相关，养阴润目丸对干眼的治疗具有一定的应用前景。

〔关键词〕干眼；养阴润目丸；结膜上皮细胞；ICAM-1；p38MAPK；p-p38MAPK

Effects of Yangyin Runmu Pills on the Expression of ICAM-1, p38MAPK and p-p38MAPK in Conjunctival Epithelial Cells of Dry Eye Model Rats

LI Dian1, WANG Chaoqun2, LIAO Liangying1, HU Ping2, PENG Yinyan2

(1. The First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China; 2. Hunan University of Chinese Medicine, Changsha, Hunan 410208, China)

〔Abstract〕Objective To study the effect of Yangyin Runmu Pills on the expression of intercellular adhesion molecule-1(ICAM-1), p38mitogen-activated Protein kinase (p38MAPK), phospho-and mitogen activated protein kinase p38antibody (pp38MAPK) in conjunctival epithelial cells of dry eye model rats. Methods Sixty Sprague -Dawley (SD) male rats were randomly divided into five groups (the normal group, sham operation group, Yangyin Runmu Pills group, Xinleiran group, model group), and 12rats in each group. All groups except for the normal group and sham operation group were prepared as the dry eye models by using the method of ovariectomy. After one week, the rats were treated with medicines. After three months treatment, schirmer tear test (STT) and tear film breakup time (BUT) of rats were detected. Theconjunctival epithelialcells were taken to test the expressive activities of p38MAPK by immunohistochemical methodand ICAM-1and p-p38MAPK by Western blot. Results Compared with the model group, SIT of Yangyin Runmu Pills group was increased obviously, BUT was extended obviously and the expressions of ICAM-1, p38MAPK, p-p38MAPK in conjunctival epithelial cellswere decreased evidently, the differenceswere statistically significant (P<0.01). Conclusion Yangyin Runmu Pills can improve the symptoms of ocular surface, increase the tear secretion,prolong appears broken, and down -regulatethe expression of ICAM-1, p38MAPK, p-p38MAPK in conjunctival epithelial cells, which suggests that the pathogenesis of dry eye is related with inflammation and p38MAPK signaling pathway, and Yangyin Runmu Pills have potential application fortreating dry eye.

〔Keywords〕dry eye；Yangyin Runmu Pills；conjunctival epithelial cell；ICAM-1；p38MAPK；p-p38MAPK

干眼，中医称为“白涩症”，又名“干涩昏花症”及“神水将枯症”,是目前最常见的眼科疾病之一。相关研究显示[1-2]在中国干眼发病率为17.0%，中国北部和西部地区的年龄超过60岁的女性群体干眼发病率高达31.3%和34.4%，而在东南地区发病率约为9.54%，且女性明显高于男性。因此探索干眼发病机制及治疗方法正逐渐成为当前的研究热点。已有研究证明[3-6]，炎症在干眼的发病机制中具有重要作用，且与MAPK信号通路激活相关。本课题组前期研究表明[7]，用经验方养阴润目丸（原名“滋阴润目丸”）治疗干眼，在改善患者主观症状及增加泪液分泌等方面具有较好的疗效，但其具体作用机制不明。……