Changes in the Expression of Smooth Muscle Contractile Proteins in TNBS- and DSS-Induced Colitis in Mice

Alkahtani, Reem; Mahavadi, Sunila; Al‐Shboul, Othman; AlSharari, Shakir D.; Grider, John R.; Murthy, Karnam S.

doi:10.1007/s10753-013-9669-0

Cited by 15 publications

(12 citation statements)

References 58 publications

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“…Colonic inflammation in mice was induced with TNBS as described previously (Hazelgrove et al, 2009;Alkahtani et al, 2013;Al-Shboul et al, 2014;Mahavadi et al, 2014;Nalli et al, 2014). Adult male mice (C57BL/6J; 6-8 weeks old) were anesthetized, and 100 ml of TNBS [2.5% in 50% ethanol (v/v)] was instilled intrarectally; mice were euthanized 3 days after the induction of inflammation.…”

Section: Methodsmentioning

confidence: 99%

Cytokine-Induced S-Nitrosylation of Soluble Guanylyl Cyclase and Expression of Phosphodiesterase 1A Contribute to Dysfunction of Longitudinal Smooth Muscle Relaxation

Rajagopal¹,

Nalli²,

Kumar³

et al. 2014

Journal of Pharmacology and Experimental Therapeutics

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The effect of proinflammatory cytokines on the expression and activity of soluble guanylyl cyclase (sGC) and cGMP-phosphodiesterases (PDEs) was determined in intestinal longitudinal smooth muscle. In control muscle cells, cGMP levels are regulated via activation of sGC and PDE5; the activity of the latter is regulated via feedback phosphorylation by cGMP-dependent protein kinase. In muscle cells isolated from muscle strips cultured with interleukin-1b (IL-1b) or tumor necrosis factor a (TNF-a) or obtained from the colon of TNBS (2,4,6-trinitrobenzene sulfonic acid)-treated mice, expression of inducible nitric oxide synthase (iNOS) was induced and sGC was S-nitrosylated, resulting in attenuation of nitric oxide (NO)-induced sGC activity and cGMP formation. The effect of cytokines on sGC S-nitrosylation and activity was blocked by the iNOS inhibitor 1400W [N-([3-(aminomethyl)phenyl]methyl)ethanimidamide dihydrochloride]. The effect of cytokines on cGMP levels measured in the absence of IBMX (3-isobutyl-1-methylxanthine), however, was partly reversed by 1400W or PDE1 inhibitor vinpocetine and completely reversed by a combination of 1400W and vinpocetine. Expression of PDE1A was induced and was accompanied by an increase in PDE1A activity in muscle cells isolated from muscle strips cultured with IL-1b or TNF-a or obtained from the colon of TNBStreated mice; the effect of cytokines on PDE1 expression and activity was blocked by, an inhibitor of nuclear factor kB activity. NO-induced muscle relaxation was inhibited in longitudinal muscle cells isolated from muscle strips cultured with IL-1b or TNF-a or obtained from the colon of TNBS-treated mice, and this inhibition was completely reversed by the combination of both 1400W and vinpocetine. Inhibition of smooth muscle relaxation during inflammation reflects the combined effects of decreased sGC activity via S-nitrosylation and increased cGMP hydrolysis via PDE1 expression.

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Section: Methodsmentioning

confidence: 99%

Cytokine-Induced S-Nitrosylation of Soluble Guanylyl Cyclase and Expression of Phosphodiesterase 1A Contribute to Dysfunction of Longitudinal Smooth Muscle Relaxation

Rajagopal¹,

Nalli²,

Kumar³

et al. 2014

Journal of Pharmacology and Experimental Therapeutics

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“…Colitis was induced with TNBS (2,4,6-trinitrobenzene sulphonic acid) as described previously (Hazelgrove et al, 2009;Alkahtani et al, 2013). Briefly, 6-to 8-week-old male adult C57BL/6J (Charles River Laboratories, Wilmington, MA) mice were fasted for 24 hours and lightly anesthesized with isoflurane; 100 ml of TNBS solution [2.5% in 50% ethanol (v/v)] was injected via a catheter advanced to 3 cm proximal to the anus via 1-ml syringe fitted with a catheter.…”

Section: Methodsmentioning

confidence: 99%

“…or TNF-a in vitro were based on our previous studies of colonic circular smooth muscle as described in Materials and Methods (Kuemmerle, 1998;Hu et al, 2007Hu et al, , 2008Hu et al, , 2009Hazelgrove et al, 2009;Alkahtani et al, 2013).…”

Section: Regulation Of Longitudinal Muscle Contraction By Cytokinesmentioning

confidence: 99%

Hypercontractility of Intestinal Longitudinal Smooth Muscle Induced by Cytokines Is Mediated by the Nuclear Factor-κB/AMP-Activated Kinase/Myosin Light Chain Kinase Pathway

Nalli¹,

Kumar²,

Mahavadi³

et al. 2014

J Pharmacol Exp Ther

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Recent studies have identified AMP-activated kinase (AMPK) as a target of Ca 21/calmodulin-dependent kinase kinase (CaMKKb) and a negative regulator of myosin light-chain (MLC) kinase (MLCK). The present study examined whether a change in expression or activity of AMPK is responsible for hypercontractility of intestinal longitudinal muscle during inflammation or in response to proinflammatory cytokines. In mouse colonic longitudinal muscle cells, acetylcholine (ACh) stimulated AMPK and MLCK phosphorylation and activity and induced MLC 20 phosphorylation and muscle contraction. Blockade of CaMKKb with STO609 (7-oxo-7H-benzimidazo[2,1-a]benz [de]isoquinoline-3-carboxylic acid acetate) inhibited AMPK and MLCK phosphorylation and augmented MLCK activity, MLC 20 phosphorylation, and smooth muscle cell contraction. In muscle cells isolated from the colon of TNBS (2,4,6-trinitrobenzenesulfonic acid)-treated mice or from strips treated with interleukin-1b or tumor necrosis factor-a, nuclear factor kB was activated as indicated by an increase in p65 phosphorylation and IkBa degradation, and AMPK was phosphorylated at a cAMP-dependent protein kinase (PKA)-specific site (Ser 485 ) that is distinct from the stimulatory CaMKKb site (Thr 172 ), resulting in attenuation of ACh-stimulated AMPK activity and augmentation of MLCK activity and muscle cell contraction. Inhibition of nuclear factor-kB activity with MG-132 (carbobenzoxy-L-leucyl-L-leucyl-L-leucinal Z-LLL-CHO) or PKA activity with myristoylated PKA inhibitor 14-22 amide blocked phosphorylation of AMPK at Ser 485 and restored MLCK activity and muscle cell contraction to control levels. The results imply that PKA released from IkBa complex phosphorylated AMPK at a PKA-specific site and inhibited its activity, thereby relieving the inhibitory effect of AMPK on MLCK and increasing MLCK activity and muscle cell contraction. We conclude that hypercontractility of intestinal longitudinal muscle induced by inflammation or proinflammatory cytokines is mediated by nuclear factor kB/PKA-dependent inhibition of AMPK and activation of MLCK.

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“…Colonic inflammation was induced with 2,4,6-trinitrobenzene sulfonic acid (TNBS) as described previously (2,14). All animal treatment protocols were approved by the Institutional Animal Care and Use Committee of Virginia Commonwealth University.…”

Section: Methodsmentioning

confidence: 99%

“…Mice were euthanized 3 days after the induction of inflammation. Colonic tissue from mice treated with TNBS exhibited typical histological characteristics of colitis (2,14). Two-to three-centimeter-long segments of the distal colon (starting from ϳ0.5 cm oral to the pelvic flexure) were removed, and muscle cells from longitudinal muscle layer were obtained.…”

Section: Methodsmentioning

confidence: 99%

Jun kinase-induced overexpression of leukemia-associated Rho GEF (LARG) mediates sustained hypercontraction of longitudinal smooth muscle in inflammation

Al‐Shboul

Nalli

Kumar

et al. 2014

American Journal of Physiology-Cell Physiology

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Al-Shboul O, Nalli AD, Kumar DP, Zhou R, Mahavadi S, Kuemmerle JF, Grider JR, Murthy KS. Jun kinase-induced overexpression of leukemia-associated Rho GEF (LARG) mediates sustained hypercontraction of longitudinal smooth muscle in inflammation. Am J Physiol Cell Physiol 306: C1129 -C1141, 2014. First published April 16, 2014; doi:10.1152/ajpcell.00021.2014.-The signaling pathways mediating sustained contraction of mouse colonic longitudinal smooth muscle and the mechanisms involved in hypercontractility of this muscle layer in response to cytokines and TNBS-induced colitis have not been fully explored. In control longitudinal smooth muscle cells, ACh acting via m3 receptors activated sequentially G␣12, RhoGEF (LARG), and the RhoA/Rho kinase pathway. There was abundant expression of MYPT1, minimal expression of CPI-17, and a notable absence of a PKC/CPI-17 pathway. LARG expression was increased in longitudinal muscle cells isolated from muscle strips cultured for 24 h with IL-1␤ or TNF-␣ or obtained from the colon of TNBS-treated mice. The increase in LARG expression was accompanied by a significant increase in ACh-stimulated Rho kinase and ZIP kinase activities, and sustained muscle contraction. The increase in LARG expression, Rho kinase and ZIP kinase activities, and sustained muscle contraction was abolished in cells pretreated with the Jun kinase inhibitor, SP600125. Expression of the MLCP activator, telokin, and MLCP activity were also decreased in longitudinal muscle cells from TNBS-treated mice or from strips treated with IL-1␤ or TNF-␣. In contrast, previous studies had shown that sustained contraction in circular smooth muscle is mediated by sequential activation of G␣13, p115RhoGEF, and dual RhoA-dependent pathways involving phosphorylation of MYPT1 and CPI-17. In colonic circular smooth muscle cells isolated from TNBS-treated mice or from strips treated with IL-1␤ or TNF-␣, CPI-17 expression and sustained muscle contraction were decreased. The disparate changes in the two muscle layers contribute to intestinal dysmotility during inflammation.colon; contraction; cytokines; signaling GASTROINTESTINAL smooth muscle exhibits regional variations in expression of structural, contractile, and signaling proteins that can influence its response to a variety of regulatory agents (e.g., acetylcholine, biogenic amines, neuropeptides, pyrimidines/purines; lipids, and cytokines) (3,12,27,37,57). Functional differences are evident between muscle layers (e.g., circular vs. longitudinal muscle of the intestine) or within the same layer (proximal tonic vs. distal rhythmic circular muscle of the stomach) (10,19,31,35,38,58,59). In all types of smooth muscle, an essential requirement for contraction is phosphorylation of the 20-kDa regulatory light chain (MLC) of myosin II (MLC 20 ), which in turn is regulated by the balance between Ca 2ϩ -dependent or -independent MLC kinase (MLCK) activity and MLC phosphatase (MLCP) activity (8,13,25,37,55).In both circular and longitudinal intestinal smooth muscle, contraction in respo...

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Changes in the Expression of Smooth Muscle Contractile Proteins in TNBS- and DSS-Induced Colitis in Mice

Cited by 15 publications

References 58 publications

Cytokine-Induced S-Nitrosylation of Soluble Guanylyl Cyclase and Expression of Phosphodiesterase 1A Contribute to Dysfunction of Longitudinal Smooth Muscle Relaxation

Cytokine-Induced S-Nitrosylation of Soluble Guanylyl Cyclase and Expression of Phosphodiesterase 1A Contribute to Dysfunction of Longitudinal Smooth Muscle Relaxation

Hypercontractility of Intestinal Longitudinal Smooth Muscle Induced by Cytokines Is Mediated by the Nuclear Factor-κB/AMP-Activated Kinase/Myosin Light Chain Kinase Pathway

Jun kinase-induced overexpression of leukemia-associated Rho GEF (LARG) mediates sustained hypercontraction of longitudinal smooth muscle in inflammation

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