Evaluation of You-Gui-Wan critical compounds inhibiting ALOX-5 and HDC gene expression in RBL-2H3 cells using a fractional factorial design

Po-Ting Chen; Pei-Chi Chen; Jiu-Yao Wang; Shulhn-Der Wang; Li-Jen Lin

doi:10.1016/j.jep.2022.116122

Evaluation of You-Gui-Wan critical compounds inhibiting ALOX-5 and HDC gene expression in RBL-2H3 cells using a fractional factorial design

J Ethnopharmacol. 2023 Apr 6:305:116122. doi: 10.1016/j.jep.2022.116122. Epub 2023 Jan 5.

Authors

Po-Ting Chen¹, Pei-Chi Chen², Jiu-Yao Wang³, Shulhn-Der Wang⁴, Li-Jen Lin⁵

Affiliations

¹ Department of Biotechnology and Food Technology, Southern Taiwan University of Science and Technology, Tainan, 71005, Taiwan. Electronic address: ptchen@stust.edu.tw.
² Center for Allergy, Immunology, and Microbiome (A.I.M.), China Medical University Hospital, Taichung, Taiwan. Electronic address: simple48686@gmail.com.
³ Center for Allergy, Immunology, and Microbiome (A.I.M.), China Medical University Hospital, Taichung, Taiwan; Children's Hospital, China Medical University, Taichung, Taiwan. Electronic address: a122@mail.ncku.edu.tw.
⁴ School of Post-Baccalaureate Chinese Medicine, College of Chinese Medicine, China Medical University, Taichung, 40402, Taiwan. Electronic address: wsd917@mail.cmu.edu.tw.
⁵ School of Chinese Medicine, College of Chinese Medicine, China Medical University, Taichung, 40402, Taiwan. Electronic address: linlijen@mail.cmu.edu.tw.

PMID: 36610671
DOI: 10.1016/j.jep.2022.116122

Abstract

Ethnopharmacological relevance: The traditional Chinese medicine (TCM) You-Gui-Wan (YGW) has been used to treat asthma for hundreds of years.

Aim of the study: YGW is composed of 10 types of medicinal materials. However, the immune mechanism of YGW in asthma treatment has not been elucidated. Therefore, this study investigated asthma symptoms attenuated by YGW and the underlying immune regulatory mechanism.

Materials and methods: Intratracheal (i.t.) stimulation of BALB/c mice with Dermatophagoides pteronyssinus (Der p) was performed once per week (40 μL, 2.5 μg/μL). For six consecutive weeks, different doses of YGW (0.2 g/kg and 0.5 g/kg) were orally administered 30 min before stimulation with Der p. After the last stimulation, airway hyperreactivity, lung gene expression, and total immunoglobulin E (IgE) in blood were evaluated using a whole-body plethysmograph system, real-time PCR, and ELISA, respectively. In addition, DNP-IgE/DNP-BSA was added to stimulate mast cells (RBL-2H3), and YGW or various compound compositions (Trial) were added to RBL-2H3 cells for 30 min to evaluate the effects of the drug on mast cell degranulation and on gene expression. JMP 5.1 software was used to design and analyze YGW's critical compounds by which it inhibited ALOX-5 and HDC gene expression in RBL-2H3 cells.

Results: YGW significantly decreased serum total IgE levels and airway hyperresponsiveness in asthmatic mice. YGW also reduced the gene expression of IL-6, TNF-α, IL-4, IL-13, and COX-2 in the lungs of asthmatic mice and RBL-2H3 cells. YGW and the compound (Trial 21) present in YGW inhibited the gene expression of ALOX-5 and HDC in RBL-2H3 cells.

Conclusion: The experimental results indicate that YGW exhibits anti-airway hyperresponsiveness and specific immunomodulatory effects. In addition, YGW synergistically inhibits ALOX-5 and HDC gene expression in mast cells through a combination of 21 compounds, including luteolin, quercetin, and β-carotene.

Keywords: Chronic asthmatic mouse model; Dermatophagoides pteronyssinus; Inflammation; Traditional Chinese medicine; You-Gui-Wan.

MeSH terms

Animals
Asthma* / drug therapy
Asthma* / genetics
Cell Degranulation
Drugs, Chinese Herbal* / pharmacology
Drugs, Chinese Herbal* / therapeutic use
Gene Expression
Immunoglobulin E
Mast Cells
Mice
Respiratory Hypersensitivity* / drug therapy

Substances

Drugs, Chinese Herbal
Immunoglobulin E
you-gui-wan
Alox5 protein, mouse