青光安颗粒剂对DBA/2J小鼠视网膜中自噬途径相关蛋白表达的影响  

Effect of Qingguangan Granules on the expression of mitophagy pathway related pro⁃teins in retinal of DBA/2J mice

作　　者：吴凯 陈立浩 时健 刘倩宏 姚小磊 WU Kai;CHEN Lihao;SHI Jian;LIU Qianhong;YAO Xiaolei(Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学,长沙410208 [2]中医药防治眼耳鼻咽喉疾病湖南省重点实验室,长沙410208 [3]河北省眼科医院,石家庄054000 [4]湖南中医药大学第一附属医院,长沙410007

出　　处：《中国中医眼科杂志》2022年第6期434-439,共6页China Journal of Chinese Ophthalmology

基　　金：国家自然科学基金地区基金项目(81860870);中国博士后科学基金面上资助项目(2018M640754);中医药防治眼耳鼻咽喉疾病湖南省重点实验室开放基金项目(2018YZD02)。

摘　　要：目的研究青光安颗粒剂(QGA)对自发性青光眼模型DBA/2J小鼠视网膜中自噬途径相关蛋白表达的影响。方法采用随机数字法将DBA/2J小鼠分为模型组(MG)、QGA低剂量组(LQGA)、QGA高剂量组(HQGA),C57BL/6J小鼠作为对照组(CG),每组10只。检测眼压并饲养38周后,连续灌胃给药15 d,取左眼视网膜。WesternBlot检测微管相关蛋白轻链3(LC3)、溶酶体相关膜蛋白1(LAMP1)、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)、细胞色素C(CytC)蛋白表达,实时荧光定量PCR(qRT-PCR)检测E3泛素连接酶(Parkin)mRNA、LAMP1 mRNA、Caspase-3 mRNA表达。结果(1)眼压:DBA/2J小鼠在第10周至第38周眼压有差异(Z=136.034,P=0.000),眼压逐渐上升,提示造模成功。(2)LAMP1:4组间小鼠视网膜LAMP1蛋白表达差异无统计学意义(P>0.05)。(3)LC3-Ⅱ/LC3-I:与MG相比,LQGA与HQGA小鼠视网膜LC3-Ⅱ/LC3-I蛋白表达降低,差异均有统计学意义(t_(LQGA)=5.140,t_(HQGA)=4.946,均P=0.000)。(4)ParkinmRNA:与MG相比,HQGA小鼠视网膜ParkinmRNA表达升高,差异有统计学意义(t=11.143,P=0.000)。(5)Caspase-3:与MG相比,LQGA、HQGA小鼠视网膜Caspase-3蛋白表达降低,差异均有统计学意义(t_(LQGA)=4.529,P=0.000;t_(HQGA)=2.764,P=0.013)。(6)CytC:与MG相比,LQGA小鼠视网膜CytC蛋白表达降低,差异有统计学意义(t=2.915,P=0.009)。结论QGA可以降低青光眼小鼠视网膜中LC3-II/LC3-I、Caspase-3的蛋白表达,一定程度上促进ParkinmRNA的表达,有利于减少自噬体的堆积和RGCs的凋亡,无明显量效关系。OBJECTIVE To investigate the effects of Qingguangan Granules(QGA)on the expression of autophagy pathway related proteins in retinal of DBA/2J mice,a model of spontaneous glaucoma.METHODS C57BL/6J mice were used as control group(CG).DBA/2J mice were divided into model group(MG),QGA low-dose group(LQGA)and QGA high-dose group(HQGA)by random number method,with ten mice in each group.Checking intraocular pressure(IOP)and after 38 weeks of feeding,the retinas on the left eyes were taken after 15 consecutive days of intragastric administration.The protein expressions of LC3-II/LC3-I,LAMP1,Caspase-3 and CytC in each group were detected by Western Blot,and the mRNA expressions of Parkin,LAMP1 and Caspase-3 were detected by qRT-PCR.RESULTS(1)IOP:The IOP of DBA/2J mice was different from the 10th week to the 38th week(Z=136.034,P=0.000),and the IOP increased gradually,indicating that modeling was successful.(2)LAMP1:There was no significant difference in LAMP1 protein´s expression in retina of mice between the 4 groups(P>0.05).(3)LC3-Ⅱ/LC3-I:Compared with MG,LC3-Ⅱ/LC3-I protein expression in retina of LQGA and HQGA mice was decreased,and the differences were statistically significant(t_(LQGA)=5.140,t_(HQGA)=4.946,all P=0.000).(4)Parkin mRNA:Compared with MG,the expression of Parkin mRNA in retina of HQGA mice was increased(t=11.143,P=0.000),and the difference was statistically significant.(5)Caspase-3:Compared with MG,the expression of retinal Caspase-3 protein in LQGA and HQGA mice was decreased,and the differences were statistically significant(t_(LQGA)=4.529,P=0.000;t_(HQGA)=2.764,P=0.013).(6)CytC:Compared with MG,the expression of CytC protein in retina of LQGA mice was decreased(t=2.915,P=0.009),and the difference was statistically significant.CONCLUSIONS QGA can reduce the protein´s expression of LC3-II/LC3-I and Caspase-3 in retinal RGCs of glaucoma mice and increase the expression of parkin mRNA to a certain extent,which is beneficial to reduce the accumulation of autophagosomes and apoptosis of RGCs.There ar

关 键 词：青光安颗粒剂 青光眼 线粒体自噬 视网膜 视网膜神经节细胞 

分 类 号：R285.5[医药卫生—中药学]