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  • Lipopolysaccharide exposure modulates the contractile and migratory phenotypes of vascular smooth muscle cells.

Lipopolysaccharide exposure modulates the contractile and migratory phenotypes of vascular smooth muscle cells.

Life sciences (2019-12-04)
Felipe Bichi Strela, Bruna Ferro Brun, Rebeca Caldeira Machado Berger, Stephano Melo, Edilamar Menezes de Oliveira, Valério Garrone Barauna, Paula Frizera Vassallo
ABSTRACT

Sepsis survivors are at higher risk for cardiovascular events. Lipopolysaccharide (LPS) activates Toll-like receptor 4 (TLR4) in sepsis. Activation of TLR4 modulates vascular smooth muscle cells (VSMCs) phenotype and contributes to cardiovascular changes after sepsis. Investigate changes in VSMCs phenotype caused by LPS-induced TLR4 activation. Rat VSMCs were incubated with LPS. Two incubation conditions were used in cell contraction and migration assays: acute stimulation - LPS stimulus was initiated at the beginning of the assay and maintained throughout; and preconditioning - LPS stimulation was applied prior to the assay then discontinued. Nitric oxide (NO) production, mRNA expression of cytokines and phenotype markers, and interleukin (IL)-6 production were evaluated. LPS increased gene expression of IL-1β, IL-6, TNFα and MCP-1 (p < .001), of secretory phenotype markers collagen and vimentin (p < .0479) and of the contractile marker smooth muscle 22α (SM22α) (p = .0067). LPS exposure increased IL-6 secretion after 24 and 48 h (p < .0001), and NO at 8 and 24 h (p < .0249) via inducible nitric oxide synthase (iNOS), as demonstrated by a decrease in NO after incubation with aminoguanidine. Acute stimulation with LPS reduced migration and contraction in a NO-dependent manner, while preconditioning with LPS increased both in an IL-6-dependent manner. LPS affects VSMCs by modulating their secretory, contractile and migratory phenotypes. LPS acute stimulation of VSMCs promoted a NO-dependent reduction in migration and contraction, while preconditioning with LPS promoted IL-6-dependent increases in migration and contraction, evidencing that VSMCs can present phenotype modifications that persist after sepsis, thereby contributing to postsepsis cardiovascular events.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Rat IL-6 ELISA Kit, for serum, plasma and cell culture supernatants
Sigma-Aldrich
Nitric oxide, 98.5%