摘要
目的通过脂多糖刺激BV2小胶质细胞建立炎症模型,探讨人参皂苷Rg1是否通过激活过氧化物酶体增殖物激活受体γ(PPARγ)受体蛋白对炎症的调控作用。方法BV2小胶质细胞随机分为对照组、模型组、人参皂苷Rg1组、罗格列酮组、GW9662组。对照组不做任何处理,模型组加入1 mg/L脂多糖,其他组在加入脂多糖处理后,再分别加入0.4 mmol/L人参皂苷Rg1、10μmol/L罗格列酮或10μmol/L GW9662。CCK-8法检测各组BV2小胶质细胞增殖情况;采用免疫荧光和免疫印迹法检测PPARγ、磷酸化核因子κBp65(p-NF-κB p65)、诱导型一氧化氮合酶(iNOS)和人精氨酸酶1(ARG-1)蛋白的表达。ELISA法检测炎症因子白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、白细胞介素8(IL-8)和肿瘤坏死因子α(TNF-α)的含量。结果与对照组相比,模型组细胞增殖率明显上升,IL-1β、IL-6、IL-8和TNF-α炎症因子含量明显增高,免疫荧光和免疫印迹结果显示,iNOS和p-NF-κB p65阳性表达明显增多,PPARγ和ARG-1阳性表达明显减少(均P<0.01);与模型组相比,人参皂苷Rg1组细胞增殖率降低,IL-1β、IL-6、IL-8和TNF-α表达水平降低,iNOS和p-NF-κB p65阳性表达明显减弱,PPARγ和ARG-1阳性表达明显增强(均P<0.01)。结论人参皂苷Rg1抑制脂多糖刺激后BV2小胶质细胞的炎症反应,其机制可能与调控PPARγ/NF-κB通路从而促进小胶质细胞M2型极化有关。
Objective To establish an inflammation model by stimulating BV2 microglia by lipopolysaccharide,and to explore the regulation effect of ginsenoside Rgl on inflammation by activating peroxisome proliferator activated receptor(PPAR-γ)receptor protein.Methods BV2 microglia were randomly divided into control group,model group,ginsenoside Rgl group,rosiglitazone group and GW9662 group.The control group did not do any treatment,the model group was treated with 1 mg/L lipopolysaccharide,and the other groups were treated with lipopolysaccharide added with 0.4 mmol/L ginsenoside Rgl,10μmol/L rosiglitazone or 10μmol/L respectively.GW9662.The proliferation of BV2 microglia in each group was detected by CCK-8 method;PPAR-γ,phospho-NF-kB p65(p-NF-kB p65),induced expression of inducible nitric oxide synthase(iNOS)and human arginase 1(ARG-1)proteins.ELISA was used to detect the inflammatory factors interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-8(IL-8)and the content of tumor necrosis factor-α(TNF-α).Results Compared with the control group,the cell proliferation rate in the model group was significantly increased,and the contents of IL-1β,IL-6,IL-8 and TNF-αincreased significantly.The results of immunofluorescence and Western blotting showed that iNOS and p-NF-kB p65 increased significantly,and the positive expressions of PPAR and ARG-1 decreased significantly(both P<0.01).The expression level of TNF-αdecreased,the positive expressions of iNOS and p-NF-kB p65 decreased significantly,and the positive expressions of PPARγ and ARG-1 increased significantly(all P<0.01).Conclusion Ginsenoside Rgl inhibits the inflammatory response of BV2 microglia after lipopolysaccharide stimulation,and its mechanism may be related to the regulation of PPAR/NF-kB pathway to promote the M2-type polarization of microglia.
作者
李婷钰
王兴航
迟晓晨
李昆芳
包翠芬
LI Ting-yu;WANG Xing-hang;CHI Xiao-chen;LI Kun-fang;BAO Cui-fen(Department of Histology and Embryology,Jinzhou Medical University,Liaoning Jinzhou 121001,China;Basic Medical Experimental Teaching Center,Jinzhou Medical University,Liaoning Jinzhou 121001,China)
出处
《解剖学报》
CAS
CSCD
北大核心
2023年第3期269-275,共7页
Acta Anatomica Sinica
基金
国家自然科学基金(81774116)
辽宁省教育厅项目(JYTJCZR2020083)。
