人参皂苷Rg1通过抑制NLRP3炎症小体途径减轻氧糖剥夺/复供后小胶质细胞炎症反应

doi:10.13418/j.issn.1001-165x.2024.1.07

摘要/Abstract

摘要： 目的探讨人参皂苷Rg1通过NLRP3炎症小体途径对小胶质细胞氧糖剥夺/复供损伤后炎症反应的影响，并进一步研究其抗炎的作用机制。方法将生长状态良好的BV-2小胶质细胞随机分为6组：无处理组（Con），氧糖剥夺/复供组（OGD/R），人参皂苷Rg1低、中、高剂量组（剂量分别为0.1、0.2、0.4 mmol/L，简称Rg1L、Rg1M、Rg1H），MCC950对照组（0.05 mmol/L，MCC950）。采用CCK8法检测细胞增殖；免疫荧光和免疫印迹法检测经不同浓度人参皂苷Rg1和MCC950作用48 h后，BV-2小胶质细胞内NLRP3炎症小体相关蛋白的表达。ELISA检测BV-2小胶质细胞培养液中炎症因子IL-1β、IL-18的表达水平。结果与Con组比较，经缺氧缺糖/复供处理的BV-2小胶质细胞胞质内可见明显的Iba-1绿色荧光表达；OGD/R处理BV-2小胶质细胞2 h后，加入不同浓度人参皂苷Rg1和MCC950培养48 h后，细胞增殖率呈现明显的下降趋势。OGD/R组呈现明显的NLRP3、ASC、Caspase-1、IL-1β和IL-18阳性表达。Rg1L、Rg1M、Rg1H 3组NLRP3蛋白表达水平显著下降，并且随着药物浓度的升高，表达越低。结果表明，与OGD/R组相比，人参皂苷Rg1和MCC950可抑制活化的BV-2小胶质细胞表达NLRP3（P<0.01）。结论人参皂苷Rg1可能通过抑制NLRP3、ASC和Caspase-1蛋白的表达、干扰NLRP3炎症小体合成，进一步抑制相关炎症因子IL-1β和IL-18的表达水平，从而抑制炎症反应。

关键词: 人参皂苷Rg1； , , 氧糖剥夺/复供； , , 小胶质细胞； , , 炎症； , , 核苷酸结合寡聚化结构域样受体蛋白3

Abstract: Objective To investigate the effect of ginsenoside Rg1 on the inflammatory response of microglia cells after oxygen-glucose deprivation/resupply injury through NLRP3 inflammasome pathway, and to further investigate its anti-inflammatory mechanism. Methods BV-2 microglia were randomly divided into six groups: No treatment group (Con), oxygen glucose deprivation/resupply group (OGD/R), ginsenoside Rg1 low, medium and high dose groups (0.1, 0.2, 0.4 mmol/L, Rg1L, Rg1M, Rg1H), MCC950 control group (0.05 mmol/L, MCC950). The cell proliferation was detected by CCK8. Immunofluorescence and western blotting were used to detect the expression of NLRP3 inflammasome-related protein in BV-2 microglia after treatment with different concentrations of ginsenoside Rg1 and MCC950 for 48 h. The expression levels of IL-1β and IL-18 in BV-2 microglia were detected by ELISA. Results Compared with Con group, Iba-1 green fluorescence was observed in the cytoplasm of BV-2 microglia cells treated with hypoxia and glucose deficiency/resupply. After OGD/R treatment of BV-2 microglia for 2h, the proliferation rate of BV-2 microglia cells showed an obvious downward trend after 48 h culture with different concentrations of ginsenoside Rg1 and MCC950. There were significant positive expressions of NLRP3, ASC, Caspase-1, IL-1β and IL-18 in the OGD/R group. The NLRP3 protein expression in Rg1L, Rg1M and Rg1H groups was significantly decreased, and the expression decreased with the increasing of drug concentration. The results showed that compared with OGD/R group, ginsenoside Rg1 and MCC950 inhibited the expression of NLRP3 in activated BV-2 microglia cells (P<0.01). Conclusions Ginsenoside Rg1 may inhibit the expression levels of IL-1β and IL-18, thus inhibiting the inflammatory response by inhibiting the expressions of NLRP3, ASC, Caspase-1 and interfering with NLRP3 inflammasome synthesis.

Key words: Ginsenoside Rg1; , , Oxygen and glucose deprivation/resupply; , , Microglia; , , Inflammation; , , NLRP3

中图分类号:

R285.5

王兴航, 丁佳媛, 李放, 包翠芬, 阎丽菁. 人参皂苷Rg1通过抑制NLRP3炎症小体途径减轻氧糖剥夺/复供后小胶质细胞炎症反应[J]. 中国临床解剖学杂志, 2024, 42(1): 33-41.

Wang Xinghang, Ding Jiayuan, Li Fang, Bao Cuifen, Yan Lijing. Ginsenoside Rg1 reduces the inflammatory response of microglia after oxygen glucose deprivation/resupply by inhibiting the NLRP3 inflammasome pathway[J]. Chinese Journal of Clinical Anatomy, 2024, 42(1): 33-41.

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