摘要
背景研究表明青光眼的眼压增高与转化生长因子-β（TGF-β）促进小梁网细胞外基质的堆积以及黏附分子CD44导致房水排出阻力增加有关。传统中药青光安颗粒剂是中医临床上常用的降跟压药物，其是否通过小梁网通路发挥作用尚不清楚。目的研究青光安颗粒剂对自发性青光眼模型DBA／2J小鼠房水动力学的影响及其作用机制。方法选取lO只眼压正常的3月龄雌性DBA／2J小鼠作为对照组，采用计算机随机数字分配法将20只9月龄自发性眼压升高的DBA／2J小鼠随机分为高眼压组和青光安组，青光安组小鼠采用2．5g／kg复方青光安颗粒剂灌胃，每天2次，连续15d，对照组和高眼压组小鼠以相同剂量生理盐水灌胃。采用经前房注入／抽吸系统进行眼压测量，分别以2．5、5．0μl／min的速率继续灌注，计算房水流畅系数（c）值和房水流出阻力（R）值。将3月龄DBA／2J小鼠60只按计算机随机数字分配法分为高剂量青光安组、中剂量青光安组、低剂量青光安组和空白对照组，每组各15只，分别用25．00、12．50和6．25g／kg青光安颗粒剂灌胃，空白对照组用等容量生理盐水灌胃，给药后7d麻醉条件下提取各组小鼠含青光安药物血清和空白对照动物血清，然后收集各组小鼠含小梁网巩膜组织进行培养，采用纤连蛋白（FN）、层黏连蛋白（LN）和神经元特异性烯醇化酶（NSE）免疫组织化学染色鉴定小梁网细胞。终质量浓度为0、5、10、20、50和100ng／ml的TGF-β处理小梁网细胞24h，采用MTT比色法检测各组细胞活性；用不同质量浓度含药血清培养经20ng／mlTGF-β处理的小梁细胞，分别于培养后24、48和72h采用ELISA法检测细胞上清液中TGF-β2受体质量浓度，采用Westernblot法检测各组小梁网细胞中CD44蛋白的相对表达量。结果对照组、高眼压组和青光安组小鼠在2．5td／min和5．0μl／min灌注
Background Researches showed that the increase of intraocular pressure （IOP） in glaucomatous eye is associated with the increasing resistance to aqueous humor outflow effects of transforming growth factor-β （TGF-β） and CDdd. Qingguangan is a traditional Chinese medicine and used to treat glaucoma. However,its mechanism of lowing-IOP effect is not elucidated. Objective This study was to investigate the lowing-IOP effect and mechanism of qingguangan granule in DBA/2J mouse,a spontaneous glaucoma model mice. Methods Ten 3 month-old female DBA/2J mice with normal IOP were chosen as control group, and 20 spontaneous ocular hypertension mice aged 9 months were randomized into high IOP group and qingguangan-treated group,with 10 mice for each group. The qingguangan （2.5 g/kg） was administered by garaging twice per day for consecutive 15 days in the qingguangan-treated group, and normal saline solution was used in the same way in the control group and high lOP group, lOP was measured by anterior chamber injection/suction system at a perfusion rate of 2.5 and 5.0 μl/min, respectively,and the coefficient of aqueous outflow facility （C value） and outflow resistance （R value） were calculated. Another 60 3-month-old DBA/2J mice were randomized into blank control group garaged with normal saline solution and high-,middle-and low-dose qingguangan groups gavaged with 25.00,12.50 and 6.25 g/kg drugs, respectively,and the mouse serum containing drugs was extracted 7 days after treatment. The scleral tissue with trabecular meshwork were obtained for the culture of trabecular meshwork cells and the cells were identified by immanohistochemistry of fibronectin （FN） ,laminin （LN） and neuronspecific enolase （NSE）. TGF-β was added into the medium for 24 hours with the final concentration of 0,5,10,20,50 and 100 ng/ml,and MMT chromatometry was employed to detect the cell vitality. The cells pre-treated with 20 ng/ml TGF-[3 were treated with different concentration of drug serum for 24,48 and 72 hours, a
出处
《中华实验眼科杂志》
CAS
CSCD
北大核心
2017年第12期1079-1084,共6页
Chinese Journal Of Experimental Ophthalmology
基金
国家自然科学基金项目（81403437）
中国博士后基金面上项目（2015M570681）
湖南省自然科学基金项目（2015JJ3097）
湖南省教育厅科研基金优秀青年项目（148137）
国家中医药管理局中医眼科学重点学科建设项目
湖南省中医五官科学重点学科建设项目
关键词
青光眼
药物治疗
中草药
小梁网
细胞培养
转化生长因子-β
黏附分子
DBA
2J小
鼠
青光安
Glaucoma/drug therapy
Traditional Chinese medicine
Trabecular meshwork
Cell,cultured
Transforming growth factor-[3
Adhesion molecule
Mice, DBA/2J
Qingguar^gan
