1.Reactive oxygen species inhibition improves the down-regulation of adiponectin in 3T3-L1 adipocytes induced by AGEs
Ning LIN ; Linli DAI ; Xiaoyong LI ; Hongmei ZHANG ; Qing SU
Chinese Journal of Endocrinology and Metabolism 2014;30(10):844-848
Objective To investigate the effects of reactive oxygen species (ROS) inhibition on the down-regulation of adiponectin (ADPN) in mouse 3T3-L1 adipocytes by advanced glycation end-products (AGEs).Methods AGEs were prepared for incubating with cell.3T3-L1 preadipocytes were cultured in vitro and differentiated into mature adipocytes.Cell differentiation and lipid accumulation were determined by oil red O staining.After being intervened with AGEs,2',7'-dichlorofluorescein diacetate (DCFH-DA) was used as a reactive oxygen species (ROS) capture agent and the fluorescent intensity of 2',7 '-dichlorofluorescein (DCF) was detected by flow cytometry.Adiponectin expression under AGEs in 3T3-L1 adipocytes pretreated with N-acetyl-L-cysteine(NAC) or not was detected by real-time fluorescent PCR and ELISA.Results The level of ROS in 3T3-L1 adipocytes treated with AGEs was increased.mRNA and protein of ADPN were down-regulated.After inhibition with ROS,mRNA and protein expressions of ADPN injured by AGEs were ameliorated.Conclusion Exposure of 3T3-L1 adipocytes to AGEs induces oxidative stress in vitro,which decreases the expression of ADPN,and causes functional impairment of adipose cells and insulin resistance.
2.Protective effect of wogonin on retinal ganglion cells in mice with glaucoma and its mechanism
Xiaoyu TIAN ; Ting LI ; Shengnan ZHANG ; Linli DAI
Recent Advances in Ophthalmology 2024;44(6):438-442
Objective To explore the protective effect of wogonin on retinal ganglion cells(RGC)in mice with glau-coma and its related mechanism.Methods Ten(10 eyes)C57BL/6J mice were included in the control group,and 40(40 eyes)DBA/2J mice were selected to prepare glaucoma models,which were divided into the model group,experiment 1,2 and 3 groups,with 10 mice per group.Mice in the control and model groups were given normal saline intragastrically,and mice in the experiment 1,2 and 3 groups were intragastrically given 50,100 and 150 mg·kg-1 wogonin,respectively.The right eyes were selected as experimental eyes.The intraocular pressure(IOP)of mice was compared among the five groups before and after 2,4 and 8 weeks of intragastric administration.The RGC number and retinal thickness of mice in each group were compared after hematoxylin-eosin staining.The levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and nitric oxide synthase(NOS)in the retinal tissues of mice were detected by colorimetry.The levels of inter-leukin(IL)-6,IL-1 β and tumor necrosis factor-α(TNF-α)in the retinal tissues of mice were detected by enzyme-linked im-munosorbent assay.The expression levels of nucleotide oligomerization domain-like receptor family 3(NLRP3),Caspase-1 and apoptosis-associated speck-like protein(ASC)in the retinal tissues of mice were detected by Western blot.Results Compared with the control group,IOP in the model group and the experiment 1,2 and 3 groups increased(all P<0.05)before intragastric administration.After 2,4 and 8 weeks of intragastric administration,IOP in the model group and the ex-periment 1,2 and 3 groups increased compared with the control group(all P<0.05);IOP in the experiment 1,2 and 3 groups decreased compared with the model group(all P<0.05);IOP in the experiment 2 and 3 groups decreased compared with the experiment 1 group(both P<0.05);IOP in the experiment 3 group decreased compared with the experiment 2 group(P<0.05).Compared with the control group,the number of RGC,retinal thickness,SOD and GSH-Px levels in the model group and the experiment 1,2 and 3 groups decreased,while the expression levels of NOS,IL-6,IL-1 β,TNF-α,and NLRP3,Caspase-1 and ASC proteins increased,and the differences were statistically significant(all P<0.05).Compared with the model group,the number of RGC,retinal thickness,SOD and GSH-Px levels in the experiment 1,2 and 3 groups increased,while the expression levels of NOS,IL-6,IL-1β,TNF-α,and NLRP3,Caspase-1 and ASC proteins decreased,and the differences were statistically significant(all P<0.05).Compared with the experiment 1 group,the number of RGC,retinal thickness,SOD and GSH-Px levels in the experiment 2 and 3 groups increased,while the expression levels of NOS,IL-6,IL-1 β,TNF-α,and NLRP3,Caspase-1 and ASC proteins decreased(all P<0.05);compared with the experi-ment 2 group,the number of RGC,retinal thickness,SOD and GSH-Px levels in the experiment 3 group increased,while the expression levels of NOS,IL-6,IL-1β,TNF-α,and NLRP3,Caspase-1 and ASC proteins decreased(all P<0.05).Conclusion Wogonin can improve the retinal thickness of mice with glaucoma and alleviate retinal oxidative damage and inflammatory reaction,which may be related to inhibiting the activation of NLRP3 inflammasomes.