Alu antisense RNA ameliorates methylglyoxal-induced human lens epithelial cell apoptosis by enhancing antioxidant defense  

作　　者：Pei-Yuan Wu Ning Ji Chong-Guang Wu Xiao-Die Wang Xin Liu Zhi-Xue Song Murad Khan Suleman Shah Ying-Hua Du Xiu-Fang Wang Li-Fang Yan 

机构地区：[1]Department of Genetics,Hebei Medical University,Hebei Key Lab of Laboratory Animal,Shijiazhuang 050017,Hebei Province,China [2]Department of Ophthalmology,Beijing Tiantan Hospital,Capital Medical University,Beijing 100070,China

出　　处：《International Journal of Ophthalmology(English edition)》2023年第2期178-190,共13页国际眼科杂志（英文版）

基　　金：Supported by the National Natural Science Foundation of China(No.81771499);the Natural Science Foundation of Hebei Province,China(No.H2018206099,No.H2021206460)。

摘　　要：AIM:To determine whether an antisense RNA corresponding to the human Alu transposable element(Aluas RNA)can protect human lens epithelial cells(HLECs)from methylglyoxal-induced apoptosis.METHODS:Cell counting kit-8(CCK-8)and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assays were used to assess HLEC viability.HLEC viability/death was detected using a Calcein-AM/PI double staining kit;the annexin V-FITC method was used to detect HLEC apoptosis.The cytosolic reactive oxygen species(ROS)levels in HLECs were determined using a reactive species assay kit.The levels of malondialdehyde(MDA)and the antioxidant activities of total-superoxide dismutase(T-SOD)and glutathione peroxidase(GSH-Px)were assessed in HLECs using their respective kits.RT-q PCR and Western blotting were used to measure m RNA and protein expression levels of the genes.RESULTS:Aluas RNA rescued methylglyoxal-induced apoptosis in HLECs and ameliorated both the methylglyoxalinduced decrease in Bcl-2 m RNA and the methylglyoxalinduced increase in Bax m RNA.In addition,Aluas RNA inhibited the methylglyoxal-induced increase in Alu sense RNA expression.Aluas RNA inhibited the production of ROS induced by methylglyoxal,restored T-SOD and GSHPx activity,and moderated the increase in MDA content after treatment with methylglyoxal.Aluas RNA significantly restored the methylglyoxal-induced down-regulation of Nrf2 gene and antioxidant defense genes,including glutathione peroxidase,heme oxygenase 1,γ-glutamylcysteine synthetase and quinone oxidoreductase 1.Aluas RNA ameliorated methylglyoxal-induced increases of the m RNA and protein expression of Keap1 that is the negative regulator of Nrf2.CONCLUSION:Aluas RNA reduces apoptosis induced by methylglyoxal by enhancing antioxidant defense.

关 键 词：human Alu antisense RNA human lens epithelial cells methylglyoxal toxicity antioxidant defense apoptosis 

分 类 号：R776.1[医药卫生—眼科]