Establishment of Poly （I∶C）-induced Cytokine Storm Model in Mice and Intervention Effect of Artesunate and Qingfei Paidu Decoction

SHI Hang; ZHOU Hongying; LI Lanfang; YU Guihua; CHENG Hui; LI Canghai; WANG Huajing

doi:10.13422/j.cnki.syfjx.20230801

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Establishment of Poly （I∶C）-induced Cytokine Storm Model in Mice and Intervention Effect of Artesunate and Qingfei Paidu Decoction

更新时间：2023-05-16

- Establishment of Poly （I∶C）-induced Cytokine Storm Model in Mice and Intervention Effect of Artesunate and Qingfei Paidu Decoction
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 12, Pages: 94-103(2023)
- 作者机构：
  
  1.安徽中医药大学药学院，合肥 230012
  2.中国中医科学院中药研究所，北京 100700
  3.中国中医科学院青蒿素研究中心，北京 100700
  4.新安医学教育部重点实验室，合肥 230012
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20230801
  CLC： R2-0;R22;R285.5;R289;R33
- Published：20 June 2023，
  
  Published Online：17 March 2023，
  
  Received：11 November 2022，
- 稿件说明：
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石航,周虹颖,李兰芳等.Poly（I∶C）诱导小鼠细胞因子风暴模型的建立及青蒿琥酯、清肺排毒汤的干预作用[J].中国实验方剂学杂志,2023,29(12):94-103.

SHI Hang,ZHOU Hongying,LI Lanfang,et al.Establishment of Poly （I∶C）-induced Cytokine Storm Model in Mice and Intervention Effect of Artesunate and Qingfei Paidu Decoction[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(12):94-103.
石航,周虹颖,李兰芳等.Poly（I∶C）诱导小鼠细胞因子风暴模型的建立及青蒿琥酯、清肺排毒汤的干预作用[J].中国实验方剂学杂志,2023,29(12):94-103. DOI： 10.13422/j.cnki.syfjx.20230801.

SHI Hang,ZHOU Hongying,LI Lanfang,et al.Establishment of Poly （I∶C）-induced Cytokine Storm Model in Mice and Intervention Effect of Artesunate and Qingfei Paidu Decoction[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(12):94-103. DOI： 10.13422/j.cnki.syfjx.20230801.

摘要

目的

观察青蒿琥酯及清肺排毒汤对病毒模拟物Poly（I∶C）诱导的小鼠细胞因子风暴（CS）动物模型的干预作用。

方法

采用84只SPF级雄性BALB/c小鼠随机分为7组，每组12只，除空白组12只外，余下的72只以15 mg·kg

-1

剂量尾静脉注射Poly（I∶C）攻击建立CS模型小鼠，并给予青蒿琥酯低、中、高剂量组（10、20、40 mg·kg

-1

）、清肺排毒汤组（2.4 g·kg

-1

）、地塞米松组（10 mg·kg

-1

）。6 h后取肺组织、支气管肺泡灌洗液（BALF），脾脏、肺脏及外周血。计算肺脏、脾脏指数；检测BALF中炎症细胞数量；苏木素-伊红（HE）染色观察小鼠肺组织病理变化；酶联免疫吸附测定法（ELISA）检测BALF中肿瘤坏死因子-

（TNF-

）、

-干扰素（IFN-

）、白细胞介素-1

（IL-1

）、IL-6含量；流式细胞术检测并分析BALF及外周血中免疫细胞表达情况。

结果

对小鼠肺脏、脾脏指数分析显示，与空白组比较，模型组肺脏、脾脏指数均有不同程度升高（

0.05）；与模型组比较，青蒿琥酯各处理组明显降低脾脏指数（

0.05），青蒿琥酯低剂量组明显降低肺脏指数（

0.05），清肺排毒汤对肺脏、脾脏指数均明显降低（

0.05）。ELISA结果显示，与空白组比较，模型组小鼠除TNF-

外，IFN-

、IL-1

、IL-6含量均明显升高（

0.05）；与模型组比较，青蒿琥酯低剂量组及清肺排毒汤组明显降低TNF-

含量（

0.05），且各药物处理组均明显降低IFN-

、IL-1

、IL-6含量（

0.05）。对BALF中炎性细胞计数发现模型组细胞数量呈下降趋势，除地塞米松组外各给药组处理后细胞数量均明显升高（

0.05）。流式细胞术检测后发现，与空白组比较，模型组外周血中CD3数量明显降低（

0.05），Ly-6G、F4/80数量明显升高（

0.05），模型组BALF中CD45、CD3、F4/80数量明显降低（

0.05），Ly-6G数量明显上升（

0.05）；与模型组比较，外周血免疫细胞检测中发现青蒿琥酯处理组和清肺排毒汤组明显升高CD45含量（

0.05），降低Ly-6G、F4/80数量（

0.05），BALF免疫细胞检测中发现可升高CD45、F4/80含量（

0.05），降低Ly-6G数量（

0.05）。

结论

青蒿琥酯及清肺排毒汤对Poly（I∶C）诱导的小鼠CS有较好的保护作用，其作用机理与有效干预机体免疫细胞紊乱有关。

Abstract

Objective

To observe the intervention effect of artesunate （ART） and Qingfei Paidu decoction （QFPD） on the mouse model of cytokine storm （CS） induced by viral mimic Poly （I∶C）.

Method

Eighty-four SPF male BALB/c mice were randomly divided into seven groups， with 12 mice in each group. Mice， except for those in the blank group （

=12）， were subjected to CS model induction by tail vein injection of Poly （I∶C） at 15 mg·kg

-1

， followed by drug treatments of low-dose ART （ART-l， 10 mg·kg

-1

）， medium-dose ART （ART-m， 20 mg·kg

-1

）， high-dose ART （ART-h， 40 mg·kg

-1

）， Qingfei Paidu Decoction （QFPD， 2.4 g·kg

-1

）， and dexamethasone （DXM， 10 mg·kg

-1

）. After 6 hours， lung tissues， bronchoalveolar lavage fluid （BALF）， spleen， lung， and peripheral blood were collected. The lung and spleen indexes were calculated and the number of inflammatory cells in BALF was detected. The pathological changes in lung tissues were observed by hematoxylin-eosin （HE） staining and the levels of tumor necrosis factor-

（TNF-

）， interferon-

（IFN-

）， interleukin-1

（IL-1

）， and IL-6 in BALF were detected by enzyme-linked immunosorbent assay （ELISA）. The expression of immune cells in BALF and peripheral blood was detected by flow cytometry.

Result

The analysis of lung and spleen indexes showed that compared with the blank group， the model group showed increased lung and spleen indexes to varying degrees （

0.05）. Compared with the model group， the ART groups showed reduced spleen index （

0.05） and the ART-l group showed reduced lung index （

0.05）. Additionally， the QFPD group showed reduced lung and spleen indexes （

0.05）. ELISA results showed that except for TNF-

， the levels of IFN-

， IL-1

， and IL-6 in the model group increased compared with those in the blank group （

0.05）. Compared with the model group， the ART-l group and the QFPD group showed reduced content of TNF-

（

0.05）， and all groups with drug intervention showed reduced content of IFN-

， IL-1

， and IL-6 （

0.05）. The number of inflammatory cells in BALF showed a downward trend in the model group， and the number of cells increased in the groups with drug intervention except for the DXM group （

0.05）. Flow cytometry showed that compared with the blank group， the model group showed decreased number of CD3 in the peripheral blood （

0.05）， increased Ly-6G and F4/80 （

0.05）， decreased expression of CD45， CD3， and F4/80 in BALF （

0.05）， and increased expressions of Ly-6G （

0.05）. Compared with the model group， the ART groups and QFPD group showed increased CD45 content in peripheral blood （

0.05）， decreased Ly-6G and F4/80 content （

0.05）， increased CD45 and F4/80 content in BALF （

0.05）， and decreased expression of Ly-6G （

0.05）.

Conclusion

ART and QFPD have a good protective effect on Poly （I∶C）-induced CS in mice， and the mechanism is related to the effective intervention in immune cell disorder.

关键词

细胞因子风暴青蒿琥酯清肺排毒汤炎症

Keywords

cytokine stormartesunateQingfei Paidu decoctioninflammation

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