Effect of Sinisan on Macrophage Polarization of RAW264.7 Cells Induced by Lipopolysaccharide

Hui-jie FAN; Zhang-bin TAN; Hong-feng LIANG; Bin LIU; Xiao-shan ZHAO; Ying-chun ZHOU

doi:10.13422/j.cnki.syfjx.20191301

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Effect of Sinisan on Macrophage Polarization of RAW264.7 Cells Induced by Lipopolysaccharide

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- Effect of Sinisan on Macrophage Polarization of RAW264.7 Cells Induced by Lipopolysaccharide
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 13, Pages: 9-14(2019)
- 作者机构：
  
  1.南方医科大学　中医药学院，广州　 510515；
  2.阳江市人民医院，广东　阳江　 529500；
  3.广州医科大学　附属第二医院，广州　 510260
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20191301
  CLC： R2-0; R22; R285.5; R289
- Received：26 November 2018，
  
  Published Online：20 March 2019，
  
  Published：05 July 2019
- 稿件说明：
移动端阅览
Hui-jie FAN, Zhang-bin TAN, Hong-feng LIANG, et al. Effect of Sinisan on Macrophage Polarization of RAW264.7 Cells Induced by Lipopolysaccharide[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(13): 9-14.
DOI：

Hui-jie FAN, Zhang-bin TAN, Hong-feng LIANG, et al. Effect of Sinisan on Macrophage Polarization of RAW264.7 Cells Induced by Lipopolysaccharide[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(13): 9-14. DOI： 10.13422/j.cnki.syfjx.20191301.

摘要

目的：

探讨四逆散(Sinisan

SNS)对脂多糖(LPS)诱导的RAW264.7巨噬细胞极化的调控作用。

方法：

RAW264.7分为5组，分别为空白组，模型组，SNS低、中、高质量浓度组(10，20，40 mg·L

－1

)；以LPS(100 μg·L

－1

)刺激的RAW264.7细胞为体外模型，使用不同质量浓度的SNS提前干预细胞，噻唑蓝(MTT)比色法检测不同质量浓度SNS对RAW264.7细胞增殖的影响；光学显微镜下观察各组细胞分化程度；酶联免疫吸附测定(ELISA)检测细胞培养基上清中M1极化因子肿瘤坏死因子-

(TNF-

)，白细胞介素-6(IL-6)，白细胞介素-1

(IL-1

)以及M2极化因子白细胞介素-10(IL-10)的含量；实时荧光定量聚合酶链式反应(Real-time PCR)检测RAW264.7细胞M1极化因子TNF-

，IL-6以及M2极化因子IL-10，精氨酸酶-1(Arg-1)的mRNA水平。

结果：

与空白组比较，模型组促进细胞增殖(

0.05)，刺激M1极化因子TNF-

，IL-6，IL-1

的释放和上调TNF-

，IL-6的mRNA含量(

0.01)，减少M2极化因子IL-10释放和IL-10，Arg-1的mRNA水平(

0.05，

0.01)。与空白组比较，SNS对RAW264.7细胞的活性没有影响。与模型组比较，SNS可抑制LPS诱导的细胞增殖(

0.05)，减少LPS刺激的细胞分化，减少M1极化因子TNF-

，IL-6，IL-1

的释放和TNF-

，IL-6的mRNA含量(

0.05，

0.01)，并增加M2极化因子IL-10释放和IL-10，Arg-1的mRNA水平(

0.05，

0.01)。

结论：

SNS可抑制LPS诱导的RAW264.7细胞炎症，其机制可能与调控巨噬细胞M1/M2表型极化平衡相关。

Abstract

Objective:

To investigate the regulatory effect of Sinisan(SNS) on the polarization of RAW264.7 macrophages induced by lipopolysaccharide (LPS).

Method:

RAW264.7 cells stimulated by LPS were used as the

in vitro

model. The cells were intervened with the different concentrations of SNS in advance. The effects of different concentrations of SNS on the proliferation of RAW264.7 cells were detected by methyl thiazolyl tetrazolium (MTT) colorimetry. The degree of cell differentiation was detected by enzyme-linked immuno sorbent assay(ELISA) method. The contents of M1 polarization factors tumor necrosis factor-

(TNF-

)

interleukin-6 (IL-6)

interleukin-1

(IL-1

) and M2 polarization factors interleukin-10 (IL-10) in cell culture supernatant were detected by ELISA method

mRNA levels of M1 polarization factors TNF-

IL-6 and M2 polarization factors IL-10

arginase-1 (Arg-1) were detected by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) method.

Result:

SNS had no effect on the cell viability of RAW264.7 cells

inhibited LPS-induced cell proliferation

decreased LPS-stimulated cell differentiation

down-regulated M1 polarizing factors TNF-

IL-6

IL-1

release and TNF-

IL-6 mRNA levels

and increased the release of IL-10 and mRNA levels of IL-10 and Arg-1.

Conclusion:

SNS inhibits the inflammation of RAW264.7 cells induced by LPS

and its mechanism may be related to the regulation of polarization balance of M1/M2 macrophages.

关键词

Keywords

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