枸杞多糖对N-甲基-D-天冬氨酸诱导的视网膜光感受器细胞损伤的保护作用  

Protective effects of Lycium barbarum polysaccharides on retinal photoreceptor cells induced by exposure to N-methyl-D-aspartate

作　　者：张芳玲 王玉红[1] 潘天鸿 张红梅[1] ZHANG Fang-ling;WANG Yu-hong;PAN Tian-hong;ZHANG Hong-mei(Department of Ophthalmology,Nanjing Hospital Affiliated to Nanjing University of Chinese Medicine,Nanjing 210003,Jiangsu Province,China;Department of Respiratory Medicine,Children's Hospital of Nanjing Medical University,Nanjing 210003,Jiangsu Province,China)

机构地区：[1]南京中医药大学附属南京医院眼科,江苏南京210003 [2]南京医科大学附属儿童医院呼吸科,江苏南京210003

出　　处：《中国临床药理学杂志》2024年第14期2058-2062,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81300323)。

摘　　要：目的探讨枸杞多糖(LBP)对N-甲基-D-天冬氨酸(NMDA)诱导的视网膜光感受器细胞损伤的保护作用,及其可能的作用机制。方法实验分为正常组、模型组、对照组和低、中、高剂量实验组。低、中、高剂量实验组分别给予0.01、0.50、2.00 mg·mL^(-1) LBP;对照组给予10μmol·L^(-1) MK-801溶液;正常组和模型组均给予等体积的完全培养基。用细胞计数试剂盒-8(CCK-8)法观察细胞存活率,用流式细胞仪检测细胞凋亡率,用钙成像观察胞内钙离子浓度,用蛋白质印迹法检测B淋巴细胞瘤-2(Bcl-2)和Bcl-2相关X蛋白(Bax)的表达水平。结果中剂量实验组、对照组、模型组和正常组的细胞存活率分别为(85.92±6.21)%、(72.33±5.45)%、(37.92±2.31)%和(97.12±5.11)%,凋亡率分别为(13.80±0.52)%、(15.22±0.51)%、(38.10±0.67)%和(5.34±0.46)%,加入NMDA后30 s钙离子相对浓度分别为3.00±0.37、4.65±0.24、27.24±0.96和1.02±0.32,Bcl-2蛋白相对表达水平分别为0.80±0.05、0.78±0.02、0.30±0.04和1.00±0.05,Bax蛋白相对表达水平分别为1.57±0.04、1.91±0.05、5.18±0.06和1.00±0.04。中剂量实验组和对照组的上述指标与模型组比较,在统计学上差异均有统计学意义(均P<0.05)。结论LBP可减缓NMDA对视网膜光感受器细胞的损伤,其作用机制可能与抑制钙离子内流及下调Bax表达、上调Bcl-2表达有关。Objective To investigate the protective effects of Lycium barbarum polysaccharide(LBP)on retinal photoreceptor cell damage caused by exposure to N-methyl-D-aspartate(NMDA)and its possible mechanism.Methods The experiment was divided into normal group,model group,control group and experimental-L,-M,-H groups.The experimental-L,-M,-H groups were given 0.01,0.50 and 2.00 mg·mL^(-1)LBP,respectively,while the control group was given 10μmol·L^(-1)MK-801 solution.The normal and model groups were given the same volume of complete culture medium.Cell counting kit-8(CCK-8)was used to measure cell survival.Flow cytometry was used to measured cell apoptosis.Calcium imaging was used to measure changes in intracellular calcium concentration.Western blot was used to determine protein levels of B-cell lymphoma-2(Bcl-2)and Bcl-2 associated X protein(Bax).Results The cell survival rates of experimental-M,control,model and normal groups were(85.92±6.21)%,(72.33±5.45)%,(37.92±2.31)%and(97.12±5.11)%;the apoptosis rates were(13.80±0.52)%,(15.22±0.51)%,(38.10±0.67)%and(5.34±0.46)%;the intracellular calcium relative concentration after NMDA was added 30 s were 3.00±0.37,4.65±0.24,27.24±0.96 and 1.02±0.32;the relative expression levels of Bcl-2 protein were0.80±0.05,0.78±0.02,0.30±0.04 and 1.00±0.05;the relative expression levels of Bax protein were 1.57±0.04,1.91±0.05,5.18±0.06 and 1.00±0.04,respectively.Compared with the model group,the above-mentioned indexes in the experimental-M and control groups had statistical significance(all P<0.05).Conclusion LBP could preclude the damage of retinal photoreceptor cells induced by exposure to NMDA,the possible mechanisms are related to inhibit the internal flow of calcium,and decrease the Bax expression and increase Bcl-2 expression at protein level.

关 键 词：枸杞多糖 凋亡 视网膜光感受器细胞 N-甲基-D-天冬氨酸 钙超载 

分 类 号：R28[医药卫生—中药学]