Differential Cellular and Molecular Effects of Butyrate and Trichostatin A on Vascular Smooth Muscle Cells

Abstract

The histone deacetylase (HDAC) inhibitors, butyrate and trichostatin A (TSA), are epigenetic histone modifiers and proliferation inhibitors by downregulating cyclin D1, a positive cell cycle regulator, and upregulating p21Cip1 and INK family of proteins, negative cell cycle regulators. Our recent study indicated cyclin D1 upregulation in vascular smooth muscle cells (VSMC) that are proliferation-arrested by butyrate. Here we investigate whether cyclin D1 upregulation is a unique response of VSMC to butyrate or a general response to HDAC inhibitors (HDACi) by evaluating the effects of butyrate and TSA on VSMC. While butyrate and TSA inhibit VSMC proliferation via cytostatic and cytotoxic effects, respectively, they downregulate cdk4, cdk6, and cdk2, and upregulate cyclin D3, p21Cip1 and p15INK4B, and cause similar effects on key histone H3 posttranslational modifications. Conversely, cyclin D1 is upregulated by butyrate and inhibited by TSA. Assessment of glycogen synthase 3-dependent phosphorylation, subcellular localization and transcription of cyclin D1 indicates that differential effects of butyrate and TSA on cyclin D1 levels are linked to disparity in cyclin D1 gene expression. Disparity in butyrate- and TSA-induced cyclin D1 may influence transcriptional regulation of genes that are associated with changes in cellular morphology/cellular effects that these HDACi confer on VSMC, as a transcriptional modulator.

Vancomycin treatment and butyrate supplementation modulate gut microbe composition and severity of neointimal hyperplasia after arterial injury

“…While systemic inflammation induced by exogenous administration of endotoxin, a product of gram-negative gut microbes, has been shown to exacerbate neointimal hyperplasia after balloon angioplasty (Danenberg et al 2002), a direct correlation between alteration in gut microbial composition and neointimal hyperplasia has not previously been reported. Finally, while other investigators have previously demonstrated that butyrate has anti-proliferative and anti-migratory effects in VSMC (Ranganna and Yatsu 1997;Ranganna et al 2000Ranganna et al , 2003Milton et al 2012;Cantoni et al 2013), the effect of butyrate on the response to arterial injury in vivo has not been studied.…”

“…, ; Milton et al. ; Cantoni et al. ), the effect of butyrate on the response to arterial injury in vivo has not been studied.…”

“…Butyrate produced in the colonic lumen is partially metabolized by colonocytes; further uptake occurs in the liver (Cummings et al 1987). Sodium butyrate has previously been shown to have anti-proliferative and anti-migratory effects on VSMC (Ranganna and Yatsu 1997;Ranganna et al 2000Ranganna et al , 2003Milton et al 2012;Cantoni et al 2013). However, while alteration in microbial communities has been associated with the etiopathology of multiple diseases including diabetes and obesity (Turnbaugh et al 2006;Cani et al 2008) and gut microbial-derived metabolites have been shown to promote atherogenesis (Wang et al 2011), a mechanistic link between gut microbial composition, microbial-derived SCFA, and neointimal hyperplasia after arterial injury has not previously been made.…”

Trichostatin A decreases the levels of MeCP2 expression and phosphorylation and increases its chromatin binding affinity

“…67 Histone phosphorylation has been shown to globally decrease its chromatin binding affinity 68,69 and TSA has been shown to downregulate several cyclin dependent kinases (CDKs). 70 We decided to analyze two of the most intensively characterized phosphorylation sites to date, S80-P and S421-P. 71 In neurons, S80 is constitutively phosphorylated in the resting state 72 and S421 is phosphorylated in response to neuronal activity. 69 We also decided to include S164-P, a recently described abundant developmental MeCP2 phosphorylation, which has been documented to lower its binding affinity to chromatin.…”

Histone deacetylase inhibitors promote eNOS expression in vascular smooth muscle cells and suppress hypoxia‐induced cell growth

“…Previous studies reported that trichostatin A, another HDI, also suppresses VSMC proliferation . However, its definite cytotoxicity restricts its application potential . Alternatively, Bur is a naturally occurring short‐chain fatty acid and physiologically exists in the intestinal system and blood.…”