数字中医药（英文）2023，Vol.6Issue(3)：307-316,10.DOI:10.1016/j.dcmed.2023.10.006
枸杞多糖通过抑制NF-κB/NLRP3通路减少视网膜色素变性小鼠感光细胞凋亡
Inhibition of photoreceptor apoptosis in mice with retinitis pigmentosa through NF-κB/NLRP3 pathway suppression with Lycium barbarum polysaccharide
摘要
Abstract
Objective To explore whether Lycium barbarum polysaccharide(LBP)can reduce the apop-tosis of retinal photoreceptor cells in retinitis pigmentosa(RP)mice by inhibiting nuclear fac-tor-kappa B(NF-κB)/NOD-like receptor thermal protein domain-associated protein 3(NL-RP3)signaling pathway. Methods(i)In vitro experiments,mouse retinal ganglion cells(661W cells)were divided into normal,model,LBP low-dose(LBP-L,40 mg/L),LBP middle-dose(LBP-M,80 mg/L),LBP high-dose(LBP-H,160 mg/L),and positive drug control(NLRP3 inhibitor,160 mg/L)groups.And the 661W cells were exposed to varying concentrations of H2O2 ranging from 50 to 400 μmol/L to determine the optimal concentration for inducing apoptosis(200 μmol/L).Then the cell viability was assessed using Cell Counting Kit-8(CCK-8),while the apoptosis rate was detected by flow cytometry;the expression of NLRP3 was detected by immunofluo-rescence;and the expression of apoptosis markers was detected by enzyme-linked im-munosorbent assay(ELISA)and Western blot(WB).(ⅱ)In vivo assays were carried out with the use of C57/BL6 and Rd10 mice.The animal experimental groups were divided into nor-mal,model,LBP-L,LBP-M,LBP-H,and NLRP3 inhibitor groups,in which the normal group was C57/BL6 mice and the other groups were Rd10 mice.Ten mice were included in each group,and the corresponding drugs were administered intragastrically for a duration of four weeks.NF-κB/NLRP3 pathway and the expression of apoptosis markers were observed by electroretinogram,histopathological examination,and WB to assess the effects of LBP on reti-nal photoreceptor cell apoptosis. Results(i)In vitro experiments,compared with the normal group,the apoptosis rate of 661W cells in model group was significantly increased(P＜0.01),and the expression levels of key proteins of NF-κB/NLRP pathway,such as NLRP3,NF-κB,p-NF-κB,and pro-apoptotic protein caspase-3,were up-regulated(P＜0.01).The rate of Bax/Bcl-2 was increased(P＜0.01),and the concentrations of interleukin(IL)-1β and tumor necrosis factor(TNF)-α were significantly increased(P＜0.01).Compared with the model group,high dose of LBP de-creased the apoptosis rate of 661W cells(P＜0.01),and down-regulated the expression levels of the key proteins of NF-κB/NLRP3 pathway,including NF-κB,NLRP3,p-NF-κB,and caspase-3(P＜0.01).The rate of Bax/Bcl-2 was decreased(P＜0.01),and the concentra-tions of IL-1β and TNF-α were decreased(P＜0.01).(ⅱ)In vivo experiments,high dose of LBP significantly increased morphological changes in the outer nuclear layer(ONL)thickness of Rd10 mice,as well as functional changes in the amplitudes of the a-wave and b-wave(P＜0.01),which also down-regulated the expression levels of NF-κB(P＜0.05),NLRP3,p-NF-κB,and caspase-3(P＜0.01),reduced the Bax/Bcl-2 rate(P＜0.01),and decreased the concentra-tions of IL-1β(P＜0.01)and TNF-α(P＜0.05). Conclusion LBP could improve both retinal morphology and function,providing protection to photoreceptors from apoptosis through the inhibition of the NF-κB/NLRP3 pathway.关键词
视网膜色素变性/枸杞多糖/细胞凋亡/NF-κB/NLRP3通路/氧化应激Key words
Retinitis pigmentosa(RP)/Lycium barbarum polysaccharide(LBP)/Apoptosis/NF-κB/NLRP3 pathway/Oxidative stress引用本文复制引用
王英,邓颖,逯晶,彭俊,周亚莎,杨毅敬,彭清华..枸杞多糖通过抑制NF-κB/NLRP3通路减少视网膜色素变性小鼠感光细胞凋亡[J].数字中医药（英文）,2023,6(3):307-316,10.基金项目
National Natural Science Foundation of China(82274341),Scientific Research Project of Traditional Chinese Medicine of Hunan Province(B2023041),and Qihuang Scholars Support Project of PENG Qinghua,the Scientific Research Project of Hunan Provincial Department of Ed-ucation(21B0391). （82274341）
