基于SIRT1观察补精益视片对自发性青光眼模型DBA/2J小鼠RGCs凋亡的干预及影响  

Based on SIRT1 to observe the intervention and effect of Bujingyishi tablets on RGCs apoptosis in DBA/2J mice of spontaneous glaucoma model

作　　者：杨凤姣 李翔[3] 刘红佶 万婧雯 易文华 Yang Fengjiao;Li Xiang;Liu Hongji;Wan Jingwen;Yi Wenhua(Chengdu University of TCM,Chengdu,Sichuan,610075;Key Laboratory of Sichuan Province Ophthalmopathy Prevention&Cure and Visual Function Protection with TCM,Chengdu,Sichuan,610075;Hospital of Chengdu University of TCM,Chengdu,Sichuan,610072)

机构地区：[1]成都中医药大学,四川成都610075 [2]中医药眼病防治与视功能保护四川省重点实验室,四川成都610075 [3]成都中医药大学附属医院,四川成都610072

出　　处：《中医眼耳鼻喉杂志》2022年第2期74-79,共6页Journal of Chinese Ophthalmology and Otorhinolaryngology

基　　金：四川省科技计划资助(No.2018JY0658);成都中医药大学“杏林学者”提升计划(No.YXRC2019009)。

摘　　要：目的观察补精益视片对自发性青光眼模型DBA/2J小鼠RGCs的SIRT1组蛋白去乙酰化酶表观遗传调控影响,探索其干预RGCs凋亡的新机制。方法24只DBA/2J小鼠随机分为模型组,补精益视片高、中、低剂量组,对照组6只C57BL/6J小鼠,对照组及模型组2ml生理盐水灌胃、给药组补精益视片混悬液2ml灌胃连续8周,观察小鼠RGCs SIRT1 mRNA、蛋白表达及生存、凋亡率。结果(1)眼压:给药组实验后与实验前及模型组相比均降低(P<0.01或P<0.05),实验前、后模型组与对照组相比均高(P<0.05);(2)Q-PCR检测结果:模型组低于对照组、给药组均高于模型组(P<0.05),高剂量组最高;(3)Western Blot检测结果:模型组低于对照组、给药组均高于模型组(P<0.05),高剂量组最高;(4)Annexin V-FITC/PI双染法流式细胞术检测结果:模型组高于对照组、给药组均低于模型组(P<0.05),高剂量组最低;(5)CCK-8检测结果:模型组低于对照组、给药组均高于模型组(P<0.05),高剂量组最高。结论补精益视片可能通过降眼压、上调SIRT1组蛋白去乙酰化酶、抑制RGCs凋亡、促进RGCs生存而实现保护青光眼视功能的作用。Objective Based on SIRT1 to observe the effect of Bujingyishi tablets on retinal ganglion cells in DBA/2J mice of spontaneous glaucoma model and explore a new molecular mechanism of histone acetylation modification that Bujingyishi tablets intervene the apoptosis of RGCs.Methods Twenty four DBA/2J mice of spontaneous glaucoma model were randomly divided into 4 groups:model group,treatment group with high,middle,low dosage of Bujingyishi tablets,six C57BL/6J mice were used as control group.Control group and model group were treated with 2ml saline,while treatment group was treated with 2ml Bujingyishi tablets suspension for 8 weeks.Q-PCR,Western Blot,CCK-8 and Annexin V-FITC/PI double staining flow cytometry were used to analysis the mRNA and protein expression level about SIRTI,survival and apoptosis rate of RGCs.Results(1)Intraocular pressure(IOP):After experiment,IOP of treatment group was lower than pre-experiment(all P<0.01)and model group(all P<0.05);model group was higher than control group before and after experiment(P<0.05).(2)Q-PCR detection of SIRT1 mRNA:model group was lower than control group(P<0.05).Treatment group was higher than model group,and the high-dosage group had the highest expression level(all P<0.05).(3)Western blot detection of SIRT1 protein expression level:Model group was decreased compared with control group and treatment group was increased compared with model group,the high-dosage group had the highest level(all P<0.05).(4)Annexin V-FITC/PI double staining flow cytometry showed the apoptosis rate of RGCs:model group was higher than control group,while treatment group was lower than model group(all P<0.05),and high-dosage group was the lowest.(5)RGCs survival rate was detected by CCK-8:model group was lower than control group(P<0.05).Treatment group was higher than model group,and the high-dosage group was the highest(all P<0.05).Conclusion Bujingyishi tablets can protect the visual function of glaucoma by decreasing IOP,upregulating SIRT1 histon.

关 键 词：补精益视片 RGCS SIRT1组蛋白去乙酰化酶 自发性青光眼模型 

分 类 号：R285.5[医药卫生—中药学]