Necroptosis Inhibition by Hydrogen Sulfide Alleviated Hypoxia-Induced Cardiac Fibroblasts Proliferation via Sirtuin 3

Abstract

Myocardial ischemia or hypoxia can induce myocardial fibroblast proliferation and myocardial fibrosis. Hydrogen sulfide (H2S) is a gasotransmitter with multiple physiological functions. In our present study, primary cardiac fibroblasts were incubated with H2S donor sodium hydrosulfide (NaHS, 50 μM) for 4 h followed by hypoxia stimulation (containing 5% CO2 and 1% O2) for 4 h. Then, the preventive effects on cardiac fibroblast proliferation and the possible mechanisms were investigated. Our results showed that NaHS reduced the cardiac fibroblast number, decreased the hydroxyproline content; inhibited the EdU positive ratio; and down-regulated the expressions of α-smooth muscle actin (α-SMA), the antigen identified by monoclonal antibody Ki67 (Ki67), proliferating cell nuclear antigen (PCNA), collagen I, and collagen III, suggesting that hypoxia-induced cardiac fibroblasts proliferation was suppressed by NaHS. NaHS improved the mitochondrial membrane potential and attenuated oxidative stress, and inhibited dynamin-related protein 1 (DRP1), but enhanced optic atrophy protein 1 (OPA1) expression. NaHS down-regulated receptor interacting protein kinase 1 (RIPK1) and RIPK3 expression, suggesting that necroptosis was alleviated. NaHS increased the sirtuin 3 (SIRT3) expressions in hypoxia-induced cardiac fibroblasts. Moreover, after SIRT3 siRNA transfection, the inhibitory effects on cardiac fibroblast proliferation, oxidative stress, and necroptosis were weakened. In summary, necroptosis inhibition by exogenous H2S alleviated hypoxia-induced cardiac fibroblast proliferation via SIRT3.

Hydrogen Sulfide Suppresses Skin Fibroblast Proliferation via Oxidative Stress Alleviation and Necroptosis Inhibition

“…Actually, a large amount of cellular contents will be released after necroptosis, such as inflammatory factors, cell metabolites, and other substances, which all further promote cell proliferation. Recently, some researchers found that H 2 S attenuated necroptosis depending on sirtuin3 enhancement to suppress excessive proliferation in hypoxia-induced cardiac fibroblasts [ 25 ]. That is to say that the mutual regulation between cell death and proliferation is very complex.…”

“…Exogenous H 2 S improved mitochondrial function, inhibited oxidative stress, and reduced necroptosis to improve diabetic cardiomyopathy [ 24 ]. Moreover, H 2 S donors inhibited necroptosis and alleviated hypoxia-induced myocardial fibroblast proliferation depending on sirtuin 3 [ 25 ]. However, whether oxidative stress and necroptosis were involved in the possible effect of H 2 S on skin fibroblast proliferation has not been known well.…”

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

“…Another research verified that exogenous H 2 S supplementation alleviated necroptosis to suppress NLR family pyrin domain‐containing protein 3 (NLRP3) inflammasome activation and attenuate diabetic cardiomyopathy via mitochondrial dysfunction improvement and oxidative stress inhibition 30 . H 2 S also inhibited oxidative stress‐induced necroptosis and attenuated cardiac fibroblast proliferation during hypoxia 35 . Our latest research confirmed that H 2 S inhibited the proliferation of human keloid skin fibroblasts by relieving oxidative stress and necroptosis 14 .…”

“…30 H 2 S also inhibited oxidative stress-induced necroptosis and attenuated cardiac fibroblast proliferation during hypoxia. 35 Our latest research confirmed that H 2 S inhibited the proliferation of human keloid skin fibroblasts by relieving oxidative stress and necroptosis. 14 From two opposite perspectives of endogenous H 2 S disturbance and exogenous H 2 S supplement, the current research proved eloquently that H 2 S was an endogenous protective factor to alleviate oxidative stress and necroptosis, and further to inhibit excessive proliferation of skin fibroblasts.…”

“…It had been found that NaHS reduced dynamin-related protein 1 (DRP1) but increased optic tropica protein 1 (OPA1) to reduce oxidative stress and further to inhibit hypoxia-induced cardiac fibroblast proliferation. 35 In carbon tetrachloride-stimulated mice, H 2 S donor GYY4137 S-sulphydrated Keap1 to improve liver function and attenuate liver fibrosis. 36 Our previous research also confirmed that NaHS pretreatment reduced the levels of superoxide anion and superoxide, increased ΔΨm and inhibited the migration of keloid fibroblasts.…”