In vitro tests to evaluate embryotoxicity and irritation of Chinese herbal medicine (Pentaherbs formulation) for atopic dermatitis

Lu Li; Xuan Mou; Hongliang Xie; Aolin Zhang; Junwei Li; Rongyun Wang; André Seid; Ling Yin Tang; Ling Wang; Ping Chung Leung; Horst Spielmann; Chi Chiu Wang; Xiaohui Fan

doi:10.1016/j.jep.2023.116149

In vitro tests to evaluate embryotoxicity and irritation of Chinese herbal medicine (Pentaherbs formulation) for atopic dermatitis

J Ethnopharmacol. 2023 Apr 6:305:116149. doi: 10.1016/j.jep.2023.116149. Epub 2023 Jan 8.

Authors

Lu Li¹, Xuan Mou², Hongliang Xie³, Aolin Zhang⁴, Junwei Li⁵, Rongyun Wang⁶, André Seid⁷, Ling Yin Tang⁸, Ling Wang⁹, Ping Chung Leung¹⁰, Horst Spielmann¹¹, Chi Chiu Wang¹², Xiaohui Fan¹³

Affiliations

¹ Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China; Innovation Center in Zhejiang University, State Key Laboratory of Component-Based Chinese Medicine, Hangzhou, 310058, China; Future Health Laboratory, Innovation Center of Yangtze River Delta, Zhejiang University, Jiaxing, 314100, China; Department of Obstetrics and Gynecology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine; Key Laboratory of Reproductive Dysfunction Management of Zhejiang Province, Hangzhou, 310016, China; Department of Obstetrics and Gynaecology; Li Ka Shing Institute of Health Sciences; School of Biomedical Sciences; and Sichuan University-Chinese University of Hong Kong Joint Reproductive Medicine Laboratory, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong; Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong. Electronic address: luciali@zju.edu.cn.
² Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China; Innovation Center in Zhejiang University, State Key Laboratory of Component-Based Chinese Medicine, Hangzhou, 310058, China. Electronic address: 3170100133@zju.edu.cn.
³ Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China; Innovation Center in Zhejiang University, State Key Laboratory of Component-Based Chinese Medicine, Hangzhou, 310058, China. Electronic address: xiehongliang2020@126.com.
⁴ Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China; Innovation Center in Zhejiang University, State Key Laboratory of Component-Based Chinese Medicine, Hangzhou, 310058, China. Electronic address: 22119174@zju.edu.cn.
⁵ Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China; Innovation Center in Zhejiang University, State Key Laboratory of Component-Based Chinese Medicine, Hangzhou, 310058, China. Electronic address: ljw18356863466@163.com.
⁶ College of Basic Medical Sciences, Zhejiang Chinese Medical University, Hangzhou, 310053, China. Electronic address: wangrongyun3@126.com.
⁷ Institut für Pharmazie (Pharmakologie und Toxikologie), Freie Universität Berlin, Berlin, Germany. Electronic address: andre.said@fu-berlin.de.
⁸ Department of Obstetrics and Gynaecology; Li Ka Shing Institute of Health Sciences; School of Biomedical Sciences; and Sichuan University-Chinese University of Hong Kong Joint Reproductive Medicine Laboratory, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong. Electronic address: ling_yin@yahoo.com.
⁹ Hangzhou TCM Hospital of Zhejiang Chinese Medical University, Hangzhou, 310053, China. Electronic address: wangling_3627@qq.com.
¹⁰ Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong. Electronic address: pingcleung@cuhk.edu.hk.
¹¹ Institut für Pharmazie (Pharmakologie und Toxikologie), Freie Universität Berlin, Berlin, Germany. Electronic address: horst.spielmann@fu-berlin.de.
¹² Department of Obstetrics and Gynaecology; Li Ka Shing Institute of Health Sciences; School of Biomedical Sciences; and Sichuan University-Chinese University of Hong Kong Joint Reproductive Medicine Laboratory, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong; College of Basic Medical Sciences, Zhejiang Chinese Medical University, Hangzhou, 310053, China. Electronic address: ccwang@cuhk.edu.hk.
¹³ Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China; Innovation Center in Zhejiang University, State Key Laboratory of Component-Based Chinese Medicine, Hangzhou, 310058, China; Future Health Laboratory, Innovation Center of Yangtze River Delta, Zhejiang University, Jiaxing, 314100, China. Electronic address: fanxh@zju.edu.cn.

PMID: 36632857
DOI: 10.1016/j.jep.2023.116149

Abstract

Ethnopharmacological relevance: Atopic dermatitis (AD) is a common chronic inflammatory skin disorder and its prevalence is increasing in the last few decades. No treatment can cure the condition. Pregnancy often worsens the clinical manifestation. There are considerable interests in Chinese Herbal Medicine (CHM) as an alternative treatment for AD. A well tolerated CHM formula (Pentaherbs formulation, PHF) has been proven efficacious in improving life quality and reducing topical corticosteroid use in children with moderate-to-severe AD. However, safety data of PHF are not available.

Aim of the study: Our study aimed to evaluate the safety of PHF and its 5 individual herbal extracts, including embryotoxicity by Embryonic Stem Cell Test (EST) and irritation by Skin Irritation Test (SIT).

Materials and methods: Quality of 5 herbal extracts of PHF was confirmed by chromatography. In EST, mouse embryonic stem cell line (D3) and mouse fibroblast cell line (3T3) were used to study potential embryotoxicity. Three endpoints were assessed by concentration-response curves after 10 days' culture: 50% inhibition of D3 differentiation into beating cardiomyocytes (ID₅₀D3), 50% cytotoxic effects on D3 (IC₅₀D3) and on fibroblasts (IC₅₀3T3). A biostatistically based prediction model (PM) was applied to predict the embryotoxic potentials of each CHM. In SIT, epidermis equivalent commercially available kits (EpiDerm™) were used, and concentration-viability curves were obtained by MTT assay to detect skin irritations of each CHM.

Results: Chemical authentication confirmed that 5 test herbal extracts contained their main active compounds. EST results indicated that the formula PHF and its individual CHMs were non-embryotoxic, except one CHM, Amur Corktree Bark (Huang Bai, Phellodendron chinense C.K.Schneid), was weakly embryotoxic. SIT results showed that cell viability was above 50% after treatment with different concentrations of all tested CHMs.

Conclusions: Our in vitro tests provided preliminary evidence for safety of the formula PHF in embryonic stem cell test and skin irritation model, but PHF shall be cautiously used in pregnant women with AD. Further studies are needed to support its clinical application as an alternative treatment for AD, especially to the patients who plan for pregnancy or at lactation stages.

Keywords: Atopic dermatitis; Chinese herbal medicine; Embryonic stem cell test; Pentaherbs formulation; Skin irritation test.

MeSH terms

Animals
Cell Line
Dermatitis, Atopic* / drug therapy
Drugs, Chinese Herbal* / pharmacology
Embryonic Stem Cells
Female
Humans
In Vitro Techniques
Mice
Pregnancy

Substances

Drugs, Chinese Herbal