谷精草缓解脂多糖诱导ARPE-19细胞损伤的机制研究OA

Objective To investigate the protective effect of Flos Eriocauli(GJC)in Chinese herbal medicine(CHM)on lipopolysac-charide(LPS)-induced damage in human retinal pigment epithelial(ARPE-19)cells and the underlying mechanism.Methods ARPE-19 cells were randomly divided into the following groups:normal(Nor),LPS,and LPS+GJC(low/medium/high concentration).Except for the Nor group,which was cultured in normal medium,all other groups were treated with 200 ng/mL LPS for 48 h.The LPS+GJC groups were co-treated with GJC at concentrations of 25,50,and 100 μg/mL,respectively.Cell viability was assessed using the CCK-8 assay after treatment with GJC at various concentrations(0.1,1,25,50,100,and 200 μg/mL).The activities of lactate dehy-drogenase(LDH)and superoxide dismutase(SOD),as well as the level of malondialdehyde(MDA),were measured using commercial kits.Intracellular reactive oxygen species(ROS)levels were detected by fluorescence staining.The expression of related proteins was analyzed by Western blot(WB).The levels of IL-6 and vascular endothelial growth factor(VEGF)in the cell culture supernatant were determined by ELISA.Results Compared with the Nor group,GJC(0.1-200 μg/mL)had no significant effect on ARPE-19 cell viabil-ity.Compared with the Nor group,the LPS group exhibited significantly increased LDH activity and MDA level,enhanced ROS fluores-cence intensity,and decreased SOD activity in the cell supernatant(P＜0.05).Additionally,the expression levels of p-NF-κB,TNF-α,Gal-3,VEGF,and vascular endothelial growth factor receptor 2(VEGFR2),as well as the secretion of IL-6 and VEGF,were sig-nificantly upregulated(P＜0.05).Compared with the LPS group,treatment with different concentrations of GJC significantly decreased LDH activity and MDA levels,elevated SOD activity,and attenuated ROS fluorescence intensity(P＜0.05).Furthermore,GJC treat-ment down-regulated the protein expression of p-NF-κB,TNF-α,Gal-3,VEGF,and VEGFR2,and reduced the levels of IL-6 and VEGF in the supernatant(P＜0.05),showing a dose-dependent effect.Conclusion GJC protects ARPE-19 cells from LPS-induced injury by alleviating oxidative stress,inhibiting NF-κB pathway activation,mitigating the inflammatory response,and reducing angio-genesis.