白藜芦醇通过上调annexin A1表达促进氧糖剥夺/复氧大鼠小胶质细胞系向M2型极化

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (5): 715-720.

白藜芦醇通过上调annexin A1表达促进氧糖剥夺/复氧大鼠小胶质细胞系向M2型极化

林敏, 张红梅^*

重庆大学附属三峡医院药学部, 重庆 404000

收稿日期:2020-05-09 修回日期:2020-10-13 出版日期:2021-05-05 发布日期:2021-05-06
通讯作者: ^*284413788@qq.com

Resveratrol promotes the polarization of rat microglial cell line to M2 type with oxygen glucose deprivation/reoxygenation via up-regulating the expression of annexin A1

LIN Min, ZHANG Hong-mei^*

Department of Pharmacy, Chongqing University Three Gorges Hospital, Chongqing 404000, China

Received:2020-05-09 Revised:2020-10-13 Online:2021-05-05 Published:2021-05-06
Contact: ^*284413788@qq.com

摘要/Abstract

摘要： 目的探索annexin A1(ANXA1)在白藜芦醇调控氧糖剥夺/复氧(OGD/R)大鼠小胶质细胞N9向M2型极化的作用。方法采用慢病毒LV-shAnxA1沉默N9细胞AnxA1表达,慢病毒LV-Scramble作为对照慢病毒,采用Western blot和RT-qPCR验证慢病毒感染效率。慢病毒转染3 d后构建N9细胞OGD/R模型,复氧24 h后采用白藜芦醇连续处理细胞24 h。分组如下:OGD/R(-)组、OGD/R(+)组、OGD/R(+)+Res组、OGD/R(+)+Res+LV-Scramble组、OGD/R(+)+Res+LV-shAnxA1。RT-qPCR和Western blot检测ANXA1、CD11b、肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、精氨酸酶-1(Arg-1)、白介素-10(IL-10)mRNA和/或蛋白含量。免疫荧光观察ANXA1在细胞内分布情况。结果 OGD/R(+)+Res组ANXA1 mRNA和蛋白含量较OGD/R(+)组明显增多。OGD/R(+)组ANXA1蛋白主要分布在细胞核,而OGD/R(+)+Res组ANXA1蛋白主要分布在细胞质。有效抑制ANXA1表达后,OGD/R(+)+Res+ LV-shAnxA1组ANXA1主要位于细胞核。与OGD/R(+)+Res组相比,OGD/R(+)+Res+LV-shAnxA1组CD11b蛋白含量明显增高(P＜0.01),TNF-α和IL-1β mRNA含量均明显增高(P＜0.001),而Arg-1和IL-10 mRNA含量均显著降低。结论白藜芦醇可通过上调ANXA1表达促进OGD/R鼠小胶质细胞系向M2型极化。

关键词: 白藜芦醇, ANXA1, 氧糖剥夺/复氧(OGD/R), 小胶质细胞, M2型极化

Abstract: Objective To explore the role of annexin A1 (ANXA1) in resveratrol regulation of M2 type polarization of rat microglial cell line with oxygen glucose deprivation/reoxygenation (OGD/R) model. Methods Lentivirus LV-shAnxA1 was used to silence the expression of AnxA1 in N9 cells. LV-Scramble acted as a control lentivirus. Western blot and RT-qPCR were used to verify the efficiency of lentivirus infection. 3 d after lentivirus infection, the N9 cell OGD/R model was constructed. 3 days after reoxygenation, the cells were treated with resveratrol for 24 hours. The groups were as follows: OGD/R(-) group, OGD/R(+) group, OGD/R(+)+Res group, OGD/R(+)+Res+LV-Scramble group, OGD/R(+)+Res+LV-shAnxA1. RT-qPCR and Western blot were used to detect the mRNA and/or protein content of ANXA1, CD11b, tumor necrosis factor (TNF-α), Interleukin-1β(IL-1β), arginase-1 (Arg-1) and Interleukin-10 (IL-10). Immunofluorescence was applied to observe the distribution of ANXA1 in cells. Results The level of ANXA1 mRNA and protein in OGD/R(+)+Res group was significantly higher than that in OGD/R(+) group. The ANXA1 protein in the OGD/R(+) group was mainly distributed in the nucleus,while the ANXA1 protein in the OGD/R(+)+Res group was mainly distributed in cytoplasm. After inhibiting the expression of ANXA1, the OGD/R(+)+Res+LV-shAnxA1 group ANXA1 was mainly located in the nucleus. Compared with the OGD/R(+)+Res group, the CD11b protein content in the OGD/R(+)+Res+LV-shAnxA1 group was significantly increased (P＜0.01), and the TNF-α and IL-1β mRNA levels were significantly increased(P＜0.001), while Arg-1 and IL-10 mRNA levels were significantly reduced. Conclusions Resveratrol promotes the polarization of rat microglial cell line wih OGD/R model to M2 type via upregulation of ANXA1 expression.

Key words: resveratrol, ANXA1, oxygen glucose deprivation/reoxygenation(OGD/R), microglia, M2 polarization

中图分类号:

R285.5

林敏, 张红梅. 白藜芦醇通过上调annexin A1表达促进氧糖剥夺/复氧大鼠小胶质细胞系向M2型极化[J]. 基础医学与临床, 2021, 41(5): 715-720.

LIN Min, ZHANG Hong-mei. Resveratrol promotes the polarization of rat microglial cell line to M2 type with oxygen glucose deprivation/reoxygenation via up-regulating the expression of annexin A1[J]. Basic & Clinical Medicine, 2021, 41(5): 715-720.

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