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J Ethnopharmacol. 2020 Jan 10;246:112246. doi: 10.1016/j.jep.2019.112246. Epub 2019 Sep 17.

Kan-Lu-Hsiao-Tu-Tan, a traditional Chinese medicine formula, inhibits human neutrophil activation and ameliorates imiquimod-induced psoriasis-like skin inflammation.

Author information

1
Graduate Institute of Clinical Medical Sciences, College of Medicine, Chang Gung University, Taoyuan, 333, Taiwan; Supervisory Board, Taoyuan Chinese Medicine Association, Taoyuan, 338, Taiwan; Puxin Fengze Chinese Medicine Clinic, Taoyuan, 326, Taiwan. Electronic address: moonlight0604@hotmail.com.
2
Graduate Institute of Clinical Medical Sciences, College of Medicine, Chang Gung University, Taoyuan, 333, Taiwan; Center for Traditional Chinese Medicine, Chang Gung Memorial Hospital, Taoyuan, 333, Taiwan; School of Traditional Chinese Medicine, Chang Gung University, Taoyuan, 333, Taiwan. Electronic address: misterarren@gmail.com.
3
Graduate Institute of Natural Products, College of Medicine, Chang Gung University, Taoyuan, 333, Taiwan. Electronic address: god100240847@gmail.com.
4
Research Center for Chinese Herbal Medicine, Research Center for Food and Cosmetic Safety, and Graduate Institute of Health Industry Technology, Chang Gung University of Science and Technology, Taoyuan, 333, Taiwan. Electronic address: mos19880822@gmail.com.
5
Graduate Institute of Natural Products, College of Medicine, Chang Gung University, Taoyuan, 333, Taiwan. Electronic address: 107sju@gmail.com.
6
Graduate Institute of Natural Products, College of Medicine, Chang Gung University, Taoyuan, 333, Taiwan. Electronic address: max869301@gmail.com.
7
Center for Traditional Chinese Medicine, Chang Gung Memorial Hospital, Taoyuan, 333, Taiwan; School of Traditional Chinese Medicine, Chang Gung University, Taoyuan, 333, Taiwan. Electronic address: dryang@mail.cgu.edu.tw.
8
Graduate Institute of Natural Products, College of Medicine, Chang Gung University, Taoyuan, 333, Taiwan; Research Center for Chinese Herbal Medicine, Research Center for Food and Cosmetic Safety, and Graduate Institute of Health Industry Technology, Chang Gung University of Science and Technology, Taoyuan, 333, Taiwan; Chinese Herbal Medicine Research Team, Healthy Aging Research Center, Chang Gung University, Taoyuan, 333, Taiwan; Department of Anesthesiology, Chang Gung Memorial Hospital, Taoyuan, 333, Taiwan; Department of Chemical Engineering, Ming Chi University of Technology, New Taipei City, 243, Taiwan. Electronic address: htl@mail.cgu.edu.tw.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE:

Kan-Lu-Hsiao-Tu-Tan (KLHTT) is a popular traditional Chinese medicine for treating various inflammatory diseases.

AIM OF THE STUDY:

The aim of the present study was to investigate the anti-inflammatory effects of KLHTT on human neutrophils and its therapeutic potential in treating imiquimod (IMQ)-induced psoriasis-like skin inflammation.

MATERIALS AND METHODS:

Spectrophotometry, flow cytometry, and microscopy with immunohistochemical staining were used to evaluate superoxide anion generation, elastase release, CD11b expression, adhesion, and neutrophil extracellular trap (NET) formation in activated human neutrophils. Reactive oxygen species (ROS) and reactive nitrogen species in cell-free systems were measured using a multi-well fluorometer or a spectrophotometer. A psoriasis-like skin inflammation was induced in mice using the IMQ cream.

RESULTS:

KLHTT suppressed superoxide anion generation, ROS production, CD11b expression, and adhesion in activated human neutrophils. In contrast, KLHTT failed to alter elastase release in activated human neutrophils. Additionally, KLHTT had an ROS-scavenging effect in the AAPH assay, but it did not scavenge superoxide anions directly in the xanthine/xanthine oxidase assay. Protein kinase C (PKC)-induced NET formation most commonly occurs through ROS-dependent mechanisms. KLHTT significantly inhibited phorbol 12-myristate 13-acetate, a PKC activator, inducing NET formation. Furthermore, topical KLHTT treatment reduced the area affected by psoriasis area and severity index (PASI) score and ameliorated neutrophil infiltration in IMQ-induced psoriasis-like skin inflammation in mice.

CONCLUSIONS:

Our data show that KLHTT has anti-neutrophilic inflammatory effects in inhibiting ROS generation and cell adhesion. KLHTT also mitigated NET formation, mainly via an ROS-dependent pathway. In addition, KLHTT reduced neutrophil infiltration and improved the severity of IMQ-induced psoriasis-like skin inflammation in mice. Therefore, KLHTT may prove to be a safe and effective psoriasis therapy in the future.

KEYWORDS:

Human neutrophils; Kan-Lu-Hsiao-Tu-Tan; Neutrophil extracellular traps; Psoriasis; Reactive oxygen species

PMID:
31539577
DOI:
10.1016/j.jep.2019.112246

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