摘要
目的:探讨杞菊地黄丸对青光眼大鼠模型视网膜结构的保护作用,并阐明其作用机制。方法:50只SD大鼠随机分为空白对照组,模型组,低、中和高剂量杞菊地黄丸组(n=10);除空白对照组外各组大鼠通过烙闭大鼠巩膜上静脉制作青光眼动物模型,低、中和高剂量杞菊地黄丸组大鼠分别灌胃给予剂量为1、2和4g·kg^(-1)的杞菊地黄丸,空白对照组和模型组大鼠分别灌胃给予等体积生理盐水,每天1次。12周后处死大鼠,取眼球制石蜡切片,行HE染色;TUNEL染色检测视网膜神经节细胞的凋亡情况,采用实时荧光定量PCR(RT-PCR)检测各组大鼠视网膜组织中B淋巴细胞瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)和含半胱氨酸的天冬氨酸蛋白水解酶3(caspase-3)mRNA表达水平,采用免疫组织化学法检测各组大鼠视网膜组织中Bcl-2、Bax和caspase-3蛋白表达水平。结果:与空白对照组比较,模型组大鼠视网膜神经纤维排列不整齐,纤维间质水肿,细胞层呈空泡样,视网膜神经节细胞数目明显减少;与模型组比较,高剂量杞菊地黄丸组大鼠眼视网膜神经节细胞数目增多,低和中剂量杞菊地黄丸组大鼠眼视网膜神经纤维细胞排列整齐且神经节细胞形态正常。与模型组比较,低、中和高剂量杞菊地黄丸组大鼠视网膜神经节细胞凋亡指数明显减少(P<0.05),Bcl-2mRNA及蛋白表达水平明显升高(P<0.05),Bax和caspase-3 mRNA及蛋白表达水平明显降低(P<0.05)。结论:杞菊地黄丸通过调控Bcl-2、Bax和caspase-3mRNA和蛋白的表达,抑制视网膜神经节细胞的凋亡,对青光眼大鼠视网膜起保护作用。
Objective:To investigate the protective effect of Qijudihuang Pill on the retinal structures of the glaucoma model rats,and to clarify its mechanism.Methods:Fifty SD rats were randomly divided into blank control group,model group,and low,medium,and high doses of Qijudihuang Pill groups(n=10).The chronic glaucoma rat models were established by using cauterization on episcleral veins in each group except blank control group.The rats in low,medium,and high doses of Qijudihuang Pill groups were given 1,2,and 4 g·kg-1 Qijudihuang Pill by gavage,while the rats in blank control group and model group were given the same volume of saline,1 time per day.After 12 weeks of administration,the morphological changes in ocular tissue were analyzed by HE staining.The apoptosis of retinal ganglion cells was detected by TUNEL staining.The expression levels of Bcl-2,Bax,and caspase-3 mRNA in the retina tissue of the rats in various groups were determined by RT-PCR.The expression levels of Bcl-2,Bax,and caspase-3 proteins in the retina tissue of the rats in various groups were determined by immunohistochemistry.Results:Compared with blank control group,the retinal nerve fibers in model group were irregularly arranged,the interfibrillar substance was edematous,the cell layer was vacuolated,and the number of retinal ganglion cells was reduced significantly.Compared with model group,the number of retinal ganglion cells of the rats in high dose of Qijudihuang Pill group was increased;the retinal nerve fiber cells in the eyes of the rats in low dose of Qijudihuang Pill group and medium dose of Qijudihuang Pill group were arranged neatly,and the morphology of ganglion cells were normal.Compared with model group,the apoptotic indexes of retinal ganglion cells in low,medium,and high doses of Qijudihuang Pill groups were decreased significantly(P<0.05);the mRNA and protein expression levels of Bcl-2 were increased significantly(P<0.05),while the mRNA and protein expression levels of Bax and caspase-3 were decreased significantly(P<0.05).Conclusion:Qijudihuang Pill can protect the retina of glaucoma rats by regulating the mRNA and protein expressions of Bcl-2,Bax,and caspase-3 and inhibiting the apoptosis of retinal ganglion cells.
作者
赵燕
张新
张剑
田石琦
常英霞
ZHAO Yan;,ZHANG Xin;ZHANG Jian;TIAN Shiqi;CHANG Yingxia(Department of Ophthalmology,First Affiliated Hospital,Hebei Medical University,Shijiazhuang 050081,China;Department of Ophthalmology,First Affiliated Hospital,Jiamusi University, Jiamusi 154002,China)
出处
《吉林大学学报(医学版)》
CAS
CSCD
北大核心
2018年第5期994-998,共5页
Journal of Jilin University:Medicine Edition
基金
河北省卫计委医学科学计划项目资助课题(20170484)