当归多糖通过促进lncRNA MEG3表达改善糖尿病视网膜病变  

Angelica polysaccharides improve diabetic retinopathy by promoting lncRNA MEG3 expression

作　　者：李能[1] 来坚[1] LI Neng;LAI Jian(Department of Ophthalmology,Hangzhou Hospital of Traditional Chinese Medical,Hangzhou 310007,Zhejiang Province,China)

机构地区：[1]杭州市中医院眼科,浙江省杭州市310007

出　　处：《眼科新进展》2025年第2期102-107,共6页Recent Advances in Ophthalmology

基　　金：浙江省公益技术研究计划/社会发展(编号:LGF21H120001);浙江省中医药科技计划(编号:2022ZA100)。

摘　　要：目的探究当归多糖对糖尿病视网膜病变(DR)的影响及可能的作用机制。方法18只C57BL/6J小鼠注射链脲佐菌素建立DR小鼠模型,随机分为当归多糖组(289 mg·kg^(-1)当归多糖干预)、阳性对照组(217 mg·kg^(-1)羟苯磺酸钙干预)、模型组(等体积生理盐水干预),每组6只,选取6只正常小鼠设为空白组(等体积生理盐水干预),每天1次,连续灌胃给药28 d。ARPE-19细胞高糖培养基(25 mmol·L^(-1)葡萄糖)培养建立DR细胞模型,设为阴性对照组(对照载体)、lncRNA MEG3组(lncRNA MEG3过表达载体)、模型组(未转染载体的DR细胞),未经25 mmol·L^(-1)葡萄糖处理的细胞设为空白组,均培养24 h。检测小鼠空腹血糖水平;HE染色观察小鼠视网膜组织病理学变化;qPCR检测小鼠视网膜组织中lncMEG3 mRNA表达水平;CCK-8实验检测ARPE-19细胞的细胞活性;ELISA实验检测小鼠视网膜组织和ARPE-19细胞中IL-1β及IL-6表达水平;Western blot实验检测小鼠视网膜组织和ARPE-19细胞中细胞焦亡相关蛋白(Caspase-1、Cleaved-Caspase-1、GSDMD、GSDMD-N、NLRP3、IL-1β及IL-18)表达水平。结果模型组小鼠血糖含量较空白组上升,当归多糖组及阳性对照组小鼠血糖含量较模型组均下降,差异均有统计学意义(均为P<0.05)。HE染色结果显示,与空白组相比,模型组小鼠视网膜组织损伤严重,细胞排列紊乱、疏松;与模型组相比,当归多糖组及阳性对照组小鼠视网膜组织排列较整齐、紧密,组织损伤减轻。空白组、模型组、当归多糖组及阳性对照组小鼠视网膜中lncMEG3 mRNA的相对表达水平分别为1.005±0.114、0.423±0.054、0.701±0.101及0.593±0.084。与模型组相比,当归多糖组及阳性对照组小鼠视网膜中lncMEG3 mRNA的表达水平均显著升高,差异均有统计学意义(均为P<0.01)。CCK-8实验检测结果显示,与阴性对照组相比,lncRNA MEG3组ARPE-19细胞在48、72 h时存活率均上升,差异均有统计学意义(均为P<0.05)Objective To investigate the effects of angelica polysaccharides on diabetic retinopathy(DR)and possible mechanisms of action.Methods DR mouse models were established by injecting streptozotocin into 18 C57BL/6J mice.The mice were then divided randomly into an Angelica polysaccharide group(given 289 mg·kg^(-1) Angelica polysaccharides),a positive control group(given 217 mg·kg^(-1) calcium hydroxybenzenesulfonate),and a model group(given an equal volume of saline),6 mice in each group.6 normal mice were selected as a blank group(given an equal volume of saline).The drug was administered by gavage once a day for 28 consecutive days.ARPE-19 cells were exposed to a high-glucose(25 mmol·L^(-1))culture medium to establish DR cell models,which were divided into a negative control group(control vector),a lncRNA MEG3 group(lncRNA MEG3 overexpression vector),and a model group(DR cells without vector transfection).Cells not treated with 25 mmol·L^(-1) glucose were taken as a blank group.All cells were cultured for 24 h.Fasting blood glucose levels were detected in mice.Hematoxylin\|Eosin(HE)staining was performed to study the histopathological changes in the retinal tissue of mice.Quantitative polymerase chain reaction(qPCR)was performed to detect the expression level of lncMEG3 mRNAs in the retinal tissue of mice.CCK-8 assay was used to measure the activity of ARPE-19 cells.Enzyme linked immunosorbent assay(ELISA)was used to detect IL-1βand IL-6 levels in mouse retinal tissues and ARPE-19 cells.The Western blot analysis was made to detect the levels of cellular pyroptosis-related proteins(including Caspase-1,Cleaved-Caspase-1,GSDMD,GSDMD-N,NLRP3,IL-1β,and IL-18)in mouse retinal tissues and ARPE-19 cells.Results Blood glucose levels in the mice of the model group were higher than those in the mice of the blank group and lower than those in the mice of Angelica polysaccharide and positive control groups(all P<0.05).HE staining results showed that compared with those of the blank group,the mice in the mode group had sig

关 键 词：当归多糖 糖尿病视网膜病变 lncRNA MEG3 细胞焦亡 

分 类 号：R774.1[医药卫生—眼科]