Study on the mechanism underlying the regulatory effects of freeze-dried powder of aqueous extract of <italic style="font-style: italic">Lonicera japonica</italic> Thunb. on the migration of rat T lymphocytes in a coculture system

SUN Song; CHAI Limin; FENG Guiyu; LI Dongyang; ZHANG Pingxin; LIU Wei; ZHANG Yingkai

doi:10.3969/j.issn.1006-2157.2023.02.014

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Study on the mechanism underlying the regulatory effects of freeze-dried powder of aqueous extract of Lonicera japonica Thunb. on the migration of rat T lymphocytes in a coculture system

Experimentmental Studies | 更新时间：2023-03-07

- Study on the mechanism underlying the regulatory effects of freeze-dried powder of aqueous extract of Lonicera japonica Thunb. on the migration of rat T lymphocytes in a coculture system
- Journal of Beijing University of Traditional Chinese Medicine Vol. 46, Issue 2, Pages: 232-241(2023)
- 作者机构：
  
  北京中医药大学东直门医院　北京　100700
- 作者简介：
  
  Prof. CHAI Limin, Ph.D., Doctoral Supervisor. Dongzhimen Hospital, Beijing University of Chinese Medicine, No.5, Haiyuncang Hutong, Dongcheng District, Beijing 100700. E-mail: liminchai@hotmail.com
- 基金信息：
  
  Second Batch of Scientific Research Project of the National TCM Clinical Research Base of State Administration of Traditional Chinese Medicine(JDZX2015191)
- DOI：10.3969/j.issn.1006-2157.2023.02.014
  CLC： R285.5
- Received：17 October 2022，
  
  Published Online：14 December 2022，
  
  Published：28 February 2023
- 稿件说明：
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SUN Song, CHAI Limin, FENG Guiyu, et al. Study on the mechanism underlying the regulatory effects of freeze-dried powder of aqueous extract of Lonicera japonica Thunb. on the migration of rat T lymphocytes in a coculture system[J]. Journal of beijing university of traditional chinese medicine, 2023, 46(2): 232-241.
DOI：

SUN Song, CHAI Limin, FENG Guiyu, et al. Study on the mechanism underlying the regulatory effects of freeze-dried powder of aqueous extract of Lonicera japonica Thunb. on the migration of rat T lymphocytes in a coculture system[J]. Journal of beijing university of traditional chinese medicine, 2023, 46(2): 232-241. DOI： 10.3969/j.issn.1006-2157.2023.02.014.

摘要

目的

探讨忍冬藤水提物冻干粉调控共培养体系中大鼠T淋巴细胞迁移的作用机制。

方法

Transwell建立大鼠外周血T淋巴细胞与大鼠滑膜细胞株RSC－364细胞共培养体系，白细胞介素－1β(25 μg/L)诱导细胞成炎性损伤模型，以忍冬藤水提物冻干粉(100、250 mg/L)作为干预措施，白细胞介素－1β抑制剂(25 μg/L)作为阳性对照。采用高效液相色谱－电喷雾质谱(HPLC-ESI/MS)检测忍冬藤水提物组分，HE染色观察滑膜细胞形态改变，实时动态细胞分析技术(RTCA)检测淋巴细胞迁移指数变化，流式细胞仪检测CD3

＋

CD4

＋

T细胞及CD3

＋

CD8

＋

T细胞百分比变化，酶联免疫吸附测定(ELISA)检测共培养体系中可溶性血管内皮黏附分子－1(sVCAM－1)、可溶性细胞间黏附分子－1(sICAM－1)浓度。

结果

HPLC-ESI/MS结果显示，忍冬藤水提物冻干粉主要成分有咖啡酸、芦丁等。忍冬藤水提物冻干粉干预后，HE染色显示滑膜细胞数目减少，细胞形态发生不同程度改变，细胞呈圆形，胞质颜色深染，核区分相对明显。与模型组相比，RTCA检测忍冬藤水提物低剂量组48 h和忍冬藤水提物高剂量组24、48 h淋巴细胞迁移指数降低(

0.05，

0.01)；与模型组相比，流式细胞仪检测忍冬藤水提物低、高剂量组24、48 h CD3

＋

CD4

＋

T细胞百分比降低(

0.01)，忍冬藤水提物高剂量组48 h CD3

＋

CD8

＋

T细胞百分比降低(

0.01)。与模型组相比，ELISA检测共培养体系中忍冬藤水提物低、高剂量组24、48 h细胞因子sVCAM－1、sICAM－1浓度均降低(

0.01)。

结论

忍冬藤水提冻物干粉能够降低sICAM－1、sVCAM－1含量，抑制淋巴细胞迁移至滑膜细胞，抑制CD4

＋

T、CD8

＋

T细胞的迁移浸润以减轻炎症反应，缓解滑膜细胞的炎性增殖，减缓和阻滞类风湿性关节炎的疾病发展过程，从而达到缓解炎症的目的。

Abstract

Objective

To investigate the mechanism underlying the regulatory effects of freeze-dried powder of aqueous extract of

Lonicera japonica

Thunb. on the migration of rat T lymphocytes in a coculture system.

Methods

Transwell was used to establish a coculture system of rat peripheral blood T lymphocytes and rat synovial cell line RSC-364 cells

and interleukin-1β (25 μg/L) was used to induce the inflammatory injury model of cells. The freeze-dried powder of aqueous extract of

Lonicera japonica

Thunb. (100 and 250 mg/L) was used as the intervention measure

and interleukin-1β inhibitor (25 μg/L) was used as the positive control. The components of aqueous extract of

Lonicera japonica

Thunb. were analyzed by high-performance liquid chromatography-electro spray ionization/mass spectrometry (HPLC-ESI/MS). HE staining was used to observe the morphological changes of synovial cells. The change of lymphocyte migration index was determined by real-time cellular analysis(RTCA). The percentage changes of CD3

CD4

T cells and CD3

CD8

T cells were detected by flow cytometry

and the concentrations of sVCAM-1 and sICAM-1 in the coculture system were detected by ELISA.

Results

The result of HPLC-ESI/MS analysis showed that the main components of freeze-dried powder of aqueous extract of

Lonicera japonica

Thunb. were caffeic acid and rutin

etc. After the intervention of freeze-dried powder of aqueous extract of

Lonicera japonica

Thunb.

HE staining showed that the number of synovial cells was decreased and the cell morphology had changed to varying degrees; the cells were round

the cytoplasm was deeply stained

and the nucleus was relatively distinct. Compared with the model group

the migration index of T lymphocytes in the aqueous extract of

Lonicera japonica

Thunb. low-dose group at 48 h and the aqueous extract of

Lonicera japonica

Thunb. high-dose group at 24 and 48 h measured by RTCA decreased (

0.05

0.01). Compared with the model group

the percentage of CD3

CD4

T cells in the aqueous extract of

Lonicera japonica

Thunb. low-dose and high-dose groups decreased at 24 and 48 h (

0.01)

and the percentage of CD3

CD8

T cells in the aqueous extract of

Lonicera japonica

Thunb. high-dose group decreased at 48 h (

0.01). Compared with the model group

ELISA result showed that the concentrations of cytokines sVCAM-1 and sICAM-1 in the aqueous extract of

Lonicera japonica

Thunb. low-dose and high-dose groups in the coculture system decreased at 24 and 48 h (

0.01).

Conclusion

The freeze-dried powder of aqueous extract of

Lonicera japonica

Thunb. can reduce the contents of sICAM-1 and sVCAM-1

inhibit the migration of lymphocytes to synovial cells

inhibit the migration and infiltration of CD4

T and CD8

T cells to reduce inflammation

alleviate the inflammatory proliferation of synovial cells

slow down and block the disease development process of rheumatoid arthritis

so as to alleviate inflammation.

关键词

Keywords

references

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Study on the mechanism underlying the regulatory effects of freeze-dried powder of aqueous extract of Lonicera japonica Thunb. on the migration of rat T lymphocytes in a coculture system

DOI：10.3969/j.issn.1006-2157.2023.02.014

摘要

Abstract

关键词

Keywords

references