Endogenous hydrogen sulphide deficiency and exogenous hydrogen sulphide supplement regulate skin fibroblasts proliferation via necroptosis

Author:

Li Ling12,Chen Xudong1,Liu Chang1,He Ziying1,Shen Qiyan1,Zhu Yue1,Wang Xin1,Cao Shuanglin1,Yang Shengju1ORCID

Affiliation:

1. Department of Dermatology, Affiliated Hospital of Nantong University Medical School of Nantong University Nantong China

2. Yancheng First Hospital, Affiliated Hospital of Nanjing University Medical School The First people's Hospital of Yancheng Yancheng China

Abstract

AbstractAn excessive proliferation of skin fibroblasts usually results in different skin fibrotic diseases. Hydrogen sulphide (H2S) is regarded as an important endogenous gasotransmitter with various functions. The study aimed to investigate the roles and mechanisms of H2S on primary mice skin fibroblasts proliferation. Cell proliferation and collagen synthesis were assessed with the expression of α‐smooth muscle actin (α‐SMA), proliferating cell nuclear antigen (PCNA), Collagen I and Collagen III. The degree of oxidative stress was evaluated by dihydroethidium (DHE) and MitoSOX staining. Mitochondrial membrane potential (ΔΨm) was detected by JC‐1 staining. Necroptosis was evaluated with TDT‐mediated dUTP nick end labelling (TUNEL) and expression of receptor‐interacting protein kinase 1 (RIPK1), RIPK3 and mixed lineage kinase domain‐like protein (MLKL). The present study found that α‐SMA, PCNA, Collagen I and Collagen III expression were increased, oxidative stress was promoted, ΔΨm was impaired and positive rate of TUNEL staining, RIPK1 and RIPK3 expression as well as MLKL phosphorylation were all enhanced in skin fibroblasts from cystathionine γ‐lyase (CSE) knockout (KO) mice or transforming growth factor‐β1 (TGF‐β1, 10 ng/mL)‐stimulated mice skin fibroblasts, which was restored by exogenous sodium hydrosulphide (NaHS, 50 μmol/L). In conclusion, endogenous H2S production impairment in CSE‐deficient mice accelerated skin fibroblasts proliferation via promoted necroptosis, which was attenuated by exogenous H2S. Exogenous H2S supplement alleviated proliferation of skin fibroblasts with TGF‐β1 stimulation via necroptosis inhibition. This study provides evidence for H2S as a candidate agent to prevent and treat skin fibrotic diseases.

Funder

National Natural Science Foundation of China

Science and Technology Project of Nantong City

Publisher

Wiley

Subject

Dermatology,Molecular Biology,Biochemistry

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