Shiunko and Chuoko, topical Kampo medicines, inhibit the expression of gehA encoding the extracellular lipase in Cutibacterium acnes

Abstract

Antimicrobial agents have been used for eradication of Cutibacterium (formerly Propionibacterium) acnes that is an exacerbation factor of the skin disease acne vulgaris. However, the use of antibiotics is associated with an increased risk of promoting the emergence of resistant bacteria and leading to skin dysbiosis. Traditional Japanese Kampo medicines, such as Keigairengyoto, are used to treat acne. However, there is incomplete understanding regarding their functional mechanism in treatment of acne. In this study, we examined the antimicrobial and anti‐lipase activity of the Kampo medicines used empirically for acne treatment. Three oral medicines, Keigairengyoto, Seijoboufuto and Jumihaidokuto, were found to inhibit the growth of C. acnes and decrease the lipase activity. Especially, Keigairengyoto caused remarkable decrease of bacterial lipase activity. Furthermore, topical medicines such as Shiunko and Chuoko significantly decreased the lipase activity in a dose‐dependent manner, without inhibiting C. acnes growth. The topical medicines were found to inhibit the expression of gehA, which codes for extracellular lipase. Our results indicate that Shiunko and Chuoko have potential as effective acne therapeutic agents, especially because they do not promote the emergence of antimicrobial‐resistant bacteria and skin dysbiosis.

Increased prevalence of doxycycline low‐susceptible Cutibacterium acnes isolated from acne patients in Japan caused by antimicrobial use and diversification of tetracycline resistance factors

“…Total RNA was extracted from C. acnes using a Blood/Cultured Cell Total RNA Mini Kit (Favorgen Biotech Co., Ping-Tung, Taiwan). 14 Reverse transcription and reverse transcription quantitative PCR (RT-qPCR) were performed using ReverTra Ace qPCR RT Master Mix with gDNA Remover (Toyobo, Osaka, Japan) and THUNDERBIRD™ SYBR qPCR MIX (Toyobo). The following specific primers were used for the analysis of tet(W) and internal standard gene oxc (cytochrome c oxidase subunit II): tet(W)-rtF (5′-CCGACAGCAAAGTGGAAACAA), tet(W)-rtR (5′-GCAACTTGCGGATACTGACC), oxc-rtF (5′-CTTG TCATCGGCGTATTCG), and oxc-rtR (5′-CACTTCAAGCGGAAGGT GA).…”

Development of skin sebum medium and inhibition of lipase activity in Cutibacterium acnes by oleic acid

“…Lipase activity was measured using the method reported by Nakase et al [ 10 ]. 4-Methylumbelliferyl oleate (4-MUO, Sigma-Aldrich, St. Louis, USA) was used as a lipase fluorescent substrate, and fluorescence intensity was measured using Varioskan Flash 2.4 (Thermo, MS, USA) with an excitation filter of 320 nm and an emission filter of 450 nm.…”

“…4-Methylumbelliferyl oleate (4-MUO, Sigma-Aldrich, St. Louis, USA) was used as a lipase fluorescent substrate, and fluorescence intensity was measured using Varioskan Flash 2.4 (Thermo, MS, USA) with an excitation filter of 320 nm and an emission filter of 450 nm. The expression of the gehA gene encoding extracellular lipase was analysed using the method reported by Nakase et al [ 10 ]. After anaerobic incubation of C. acnes ATCC11828 for 72 hr, total RNA was extracted using a High Pure RNA Isolation Kit (Roche).…”

“…C. acnes strains with lipase activity metabolize triglycerides, producing fatty acids that play a role in inducing inflammation. Therefore, C. acnes pathogenicity has been evaluated as a contributory factor to acne vulgaris, was used to evaluate C. acnes pathogenicity [9,10]. However, the in vitro culture medium is rich in nutrients, whereas the skin environment is poor in nutrients and sebum.…”

“…acnes pathogenicity has been evaluated as a contributory factor to acne vulgaris, was used to evaluate C. acnes pathogenicity [9, 10]. However, the in vitro culture medium is rich in nutrients, whereas the skin environment is poor in nutrients and sebum.…”