Premium
Cystathionine β‐synthase and cystathionine γ‐lyase double gene transfer ameliorated homocysteine‐mediated mesangial inflammation through hydrogen sulfide generation
Author(s) -
Sen Utpal,
Qipshidze Natia,
Abe Oluwasegun,
Metreveli Naira,
Tyagi Suresh
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.590.6
Subject(s) - cystathionine beta synthase , cystathionine gamma lyase , chemistry , mapk/erk pathway , phosphorylation , transfection , downregulation and upregulation , inflammation , signal transduction , microbiology and biotechnology , biochemistry , biology , enzyme , gene , immunology , cysteine
In the present study we investigated the in vitro signaling mechanism of Hcy‐mediated inflammation and determined the regulatory role of hydrogen sulfide (H 2 S) in this process. Mesangial cells were incubated with Hcy (75 μM) and supplemented without or with H 2 S (50 μM) for 48h. Inflammatory molecules MCP‐1 and MIP‐2 were measured by ELISA in the cultured supernatant. In the Hcy treated cells, H 2 S was measured in the supernatant. Hcy‐mediated phosphorylation of JNK and ERK were measured by Western blot. To enhance the endogenous production of H 2 S and clearance of Hcy, cystathionine β‐synthase (CBS) and cystathionine γ‐lyase (CSE) genes were delivered to the cells. Our results showed that Hcy upregulated inflammatory molecules MCP‐1 and MIP‐2; whereas the endogenous production of H 2 S was attenuated. H 2 S treatment as well as CBS and CSE double gene transfer markedly reduced Hcy‐induced upregulation of MCP‐1 and MIP‐2 in the culture supernatant. Hcy phosphorylated JNK and ERK, and H 2 S supplementation as well as CBS and CSE double gene transfection attenuated Hcy‐induced phosphorylation of these two pathways. Similarly, the inhibition of JNK and ERK phosphorylation by pharmacological and siRNA blockers attenuated MCP‐1 and MIP‐2 expressions induced by Hcy. We conclude that H 2 S plays a regulatory role in Hcy‐induced inflammatory mechanism and that JNK and ERK are the two signaling pathways involved in this process.