西洋参茎叶总皂苷抑制氧糖剥夺/复氧引起的BV⁃2小胶质细胞炎性活化作用及其机制研究

刘静; 陈希; 金宏飞; 张英辉

doi:10.16306/j.1008-861x.2023.02.002

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西洋参茎叶总皂苷抑制氧糖剥夺/复氧引起的BV⁃2小胶质细胞炎性活化作用及其机制研究

中医药抗炎专栏 | 更新时间：2023-03-24

- 西洋参茎叶总皂苷抑制氧糖剥夺/复氧引起的BV⁃2小胶质细胞炎性活化作用及其机制研究
- Study on inhibitory effect and mechanism of Panax quinquefolius saponin from stems and leaves on inflammatory activation of mouse BV⁃2 microglia induced by oxygen⁃glucose deprivation/reoxygenation
- 上海中医药大学学报 2023年37卷第2期页码：7-13
- 作者机构：
  
  1.北部战区总医院检验科（辽宁　沈阳　110016）
- 作者简介：
  
  刘静，女，硕士，副主任技师，主要从事分子生物学研究
  张英辉，主管技师；E-mail：1204542997@qq.com
- 基金信息：
  
  辽宁省自然科学基金资助项目(2022-MS-046)
- DOI：10.16306/j.1008-861x.2023.02.002
  中图分类号：
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刘静,陈希,金宏飞等.西洋参茎叶总皂苷抑制氧糖剥夺/复氧引起的BV⁃2小胶质细胞炎性活化作用及其机制研究[J].上海中医药大学学报,2023,37(02):7-13.

LIU Jing,CHEN Xi,JIN Hongfei,et al.Study on inhibitory effect and mechanism of Panax quinquefolius saponin from stems and leaves on inflammatory activation of mouse BV⁃2 microglia induced by oxygen⁃glucose deprivation/reoxygenation[J].Academic Journal of Shanghai University of Traditional Chinese Medicine,2023,37(02):7-13.
刘静,陈希,金宏飞等.西洋参茎叶总皂苷抑制氧糖剥夺/复氧引起的BV⁃2小胶质细胞炎性活化作用及其机制研究[J].上海中医药大学学报,2023,37(02):7-13. DOI： 10.16306/j.1008-861x.2023.02.002.

LIU Jing,CHEN Xi,JIN Hongfei,et al.Study on inhibitory effect and mechanism of Panax quinquefolius saponin from stems and leaves on inflammatory activation of mouse BV⁃2 microglia induced by oxygen⁃glucose deprivation/reoxygenation[J].Academic Journal of Shanghai University of Traditional Chinese Medicine,2023,37(02):7-13. DOI： 10.16306/j.1008-861x.2023.02.002.

摘要

目的,2,研究西洋参茎叶总皂苷（PQS）对氧糖剥夺/复氧（OGD/R）引起的小鼠BV-2小胶质细胞炎性活化的抑制作用，并初步阐明其作用机制。,方法,2,BV-2细胞分为对照组、OGD/R组、PQS各剂量+OGD/R组。对照组细胞在常规条件下培养；OGD/R组细胞在OGD/R条件下培养；PQS各剂量+OGD/R组细胞给予PQS（30、60、90 μg/mL）预处理后，转入OGD/R条件培养。通过酶联免疫法检测上清液中炎性因子肿瘤坏死因子-α（TNF-α）、白介素-1β（IL-1β）和白介素-6（IL-6）的含量，通过CCK-8法检测PQS对OGD/R条件下细胞增殖活性的影响，流式细胞术检测胶质细胞活化标志蛋白离子钙结合适配器分子1（Iba-1）和酸性溶霉菌糖蛋白（CD68）的表达，Western blot检测核因子-κB（NF-κB）p65磷酸化及其抑制因子NF-κB抑制蛋白α（IκB-α）和NF-κB抑制因子（NKRF）蛋白的表达。,结果,2,OGD/R诱导能够明显增加BV-2细胞上清液中炎性因子TNF-α、IL-1β和IL-6含量（,P,<,0.01），增强BV-2细胞对数期增殖活性（,P,<,0.01），上调CD68表达和p-NF-κB p65（,P,<,0.01），同时下调Iba-1和NKRF表达（,P<,0.01）。PQS预处理能够有效逆转BV-2细胞中OGD/R引起的上述指标变化趋势，且其作用呈浓度依赖性（,P,<,0.05，,P<,0.01）。此外，各组BV-2细胞中IκB-α蛋白表达无明显差异（,P,>,0.05）。,结论,2,PQS能够有效抑制OGD/R引起的BV-2细胞炎性活化，其作用机制可能与上调NKRF蛋白表达有关。

Abstract

Objective： To study the inhibitory effect of Panax quinquefolium saponin from stems and leaves （PQS） on the inflammatory activation of mouse BV-2 microglia induced by oxygen-glucose deprivation/reoxygenation （OGD/R）， and to elucidate its mechanism.,Methods,2,BV-2 cells were divided into the control group， OGD/R group and PQS low， medium， high dose+OGD/R group. The cells in control group were cultured under conventional condition； the cells in OGD/R group were cultured under OGD/R condition； the cells in PQS low， medium， high dose+OGD/R group were pretreated with PQS （30， 60， 90 μg/mL）， then cultured in OGD/R condition. The cells were cultured for 48 h， then the contents of inflammatory factors such as tumor necrosis factor-α （TNF-α）， interleukin-1β（IL-1β） and IL-6 in the cell supernatant were detected by ELISA. The proliferative activity was detected by CCK-8 method， the expression of glial activation marker protein ion calcium-binding adapter molecule 1 （Iba-1） and acid lysozyme glycoprotein （CD68） were detected by flow cytometry， and the phosphorylation of nuclear factor-κB （NF-κB） p65 and its inhibitory factors NF-κB inhibitory protein α （IBK-α） as well as NF-κB inhibitory factor （NKRF） were detected by Western blot.,Results,2,OGD/R induction could significantly increase the contents of inflammatory factors such as TNF-ɑ， IL-1β and IL-6 in the supernatant of BV-2 cells （,P,<,0.01）， enhance the proliferation activity of BV-2 cells （,P,<,0.01）， up-regulate the expression of CD68 and the phosphorylation of NF-κB-p65 （,P,<,0.01）， and down-regulate the expression of Iba-1 and NKRF （,P,<,0.01）. PQS pretreatment could effectively reverse the change trend of the above indexes caused by OGD/R in BV-2 cells， and the effects were concentration-gradient dependent （,P,<,0.05，,P,<,0.01）. In addition， there was no significant difference in the expression of IκB-α protein in BV-2 cells among all groups （,P,>,0.05）.,Conclusion,2,PQS can inhibit the inflammatory activation of BV-2 cells induced by OGD/R， and its mechanism may be related to the up-regulation of NKRF protein expression.

关键词

西洋参茎叶皂苷氧糖剥夺/复氧小胶质细胞炎性活化NKRF

Keywords

Panax quinquefolius saponin from stems and leavesOGD/Rmicrogliainflammatory activationNKRF

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