Cell culture retains contractile phenotype but epigenetically modulates cell-signaling proteins of excitation-contraction coupling in colon smooth muscle cells

Shi, Xuan–Zheng; Sarna, Sushil K.

doi:10.1152/ajpgi.00369.2012

Cited by 9 publications

(13 citation statements)

References 36 publications

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“…The culture medium was changed every 3 days. Immunofluorescence staining showed that more than 95% of the cultured cells stained positive for smooth muscle‐specific α‐actin …”

Section: Methodsmentioning

confidence: 99%

Role of brain‐derived neurotrophic factor in the pathogenesis of distention‐associated abdominal pain in bowel obstruction

Lin

Winston

et al. 2018

Neurogastroenterology Motil

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“…The culture medium was changed every 3 days. Immunofluorescence staining showed that more than 95% of the cultured cells stained positive for smooth muscle‐specific α‐actin …”

Section: Methodsmentioning

confidence: 99%

Role of brain‐derived neurotrophic factor in the pathogenesis of distention‐associated abdominal pain in bowel obstruction

Lin

Winston

et al. 2018

Neurogastroenterology Motil

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“…Rat colonic circular SMCs (RCCSMC) were isolated as described previously [22, 38–41]. In brief, the circular muscle tissue pieces in 0.5 × 0.5 cm 2 size were incubated in sterile HEPES buffer (in mmol/l: 120 NaCl, 2.6 KH 2 SO 4 , 4 KCl, 2 CaCl 2 , 0.6 MgCl 2 , 25 HEPES, 14 glucose, and 2.1% essential amino acid mixture, pH 7.4) with 1.5 mg/ml collagenase (type II, 319 U/mg; Worthington, Freehold, NJ) and 1.0 mg/ml soybean trypsin inhibitor (Sigma-Aldrich) for 45 min at 31°C.…”

Section: Methodsmentioning

confidence: 99%

“…The culture medium was changed every 3 days. Immunofluorescence staining showed that more than 95% of the cultured cells stained positive for smooth muscle-specific α-actin [22, 38–41]. …”

Section: Methodsmentioning

confidence: 99%

Pathogenesis of abdominal pain in bowel obstruction: role of mechanical stress-induced upregulation of nerve growth factor in gut smooth muscle cells

Lin¹,

Winston³

et al. 2017

Pain

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Abdominal pain is one of the major symptoms in bowel obstruction (BO); its cellular mechanisms remain incompletely understood. We tested the hypothesis that mechanical stress in obstruction upregulates expression of nociception mediator nerve growth factor (NGF) in gut smooth muscle cells (SMC), and NGF sensitizes primary sensory nerve to contribute to pain in BO. Partial colon obstruction was induced with a silicon band implanted in the distal bowel of Sprague-Dawley rats. Colon-projecting sensory neurons in the dorsal root ganglia (DRG, T13 to L2) were identified for patch clamp and gene expression studies. Referred visceral sensitivity was assessed by measuring withdrawal response to stimulation by von Frey filaments (VFF) in the lower abdomen. Membrane excitability of colon-projecting DRG neurons was significantly enhanced, and the withdrawal response to VFF stimulation markedly increased in BO rats. The expression of NGF mRNA and protein was increased in a time-dependent manner (day 1 to day 7) in colonic SMC, but not in mucosa/submucosa of the obstructed colon. Mechanical stretch in vitro caused robust NGF mRNA and protein expression in colonic SMC. Treatment with anti-NGF antibody attenuated colon neuron hyper-excitability and referred hypersensitivity in BO rats. Obstruction led to significant increases of tetrodotoxin-resistant (TTX-r) Na+ currents and mRNA expression of Nav1.8, but not Nav1.6 and Nav1.7 in colon neurons; these changes were abolished by anti-NGF treatment. In conclusion, mechanical stress-induced upregulation of NGF in colon SMC underlies the visceral hypersensitivity in BO through increased gene expression and activity of TTX-resistant Na+ channels in sensory neurons.

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“…Full-thickness tissue was dissected from the disease-free margins of the resected segments. The cultures of colonic SMCs retained their contractile phenotype (41)(42)(43). Two successive digestions with papain and collagenase, as described previously by Shi and Sarna (38,41,42), were used to disperse SMCs.…”

Section: Isolation Of Hccsmcsmentioning

confidence: 99%

Colon distention induces persistent visceral hypersensitivity by mechanotranscription of pain mediators in colonic smooth muscle cells

Lin

et al. 2015

American Journal of Physiology-Gastrointestinal and Liver Physi

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Abdominal pain and distention are major complaints in irritable bowel syndrome. Abdominal distention is mainly attributed to intraluminal retention of gas or solid contents, which may cause mechanical stress to the gut wall. Visceral hypersensitivity (VHS) may account for abdominal pain. We sought to determine whether tonic colon distention causes persistent VHS and if so whether mechanical stress-induced expression (mechanotranscription) of pain mediators in colonic smooth muscle cells (SMCs) plays a role in VHS. Human colonic SMCs were isolated and stretched in vitro to investigate whether mechanical stress upregulates expression of the pain mediator cyclooxygenase-2 (COX-2). Rat colon was distended with a 5-cm-long balloon, and gene expression of COX-2, visceromotor response (VMR), and sensory neuron excitability were determined. Static stretch of colonic SMCs induced marked expression of COX-2 mRNA and protein in a force- and time-dependent manner. Subnoxious tonic distention of the distal colon at ∼30-40 mmHg for 20 or 40 min induced COX-2 expression and PGE2 production in colonic smooth muscle, but not in the mucosa layer. Lumen distention also increased VMR in a force- and time-dependent manner. The increase of VMR persisted for at least 3 days. Patch-clamp experiments showed that the excitability of colon projecting sensory neurons in the dorsal root ganglia was markedly augmented, 24 h after lumen distention. Administration of COX-2 inhibitor NS-398 partially but significantly attenuated distention-induced VHS. In conclusion, tonic lumen distention upregulates expression of COX-2 in colonic SMC, and COX-2 contributes to persistent VHS.

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Cell culture retains contractile phenotype but epigenetically modulates cell-signaling proteins of excitation-contraction coupling in colon smooth muscle cells

Abstract: Shi XZ, Sarna SK. Cell culture retains contractile phenotype but epigenetically modulates cell-signaling proteins of excitation-contraction coupling in colon smooth muscle cells.

Cited by 9 publications

References 36 publications

Role of brain‐derived neurotrophic factor in the pathogenesis of distention‐associated abdominal pain in bowel obstruction

Role of brain‐derived neurotrophic factor in the pathogenesis of distention‐associated abdominal pain in bowel obstruction

Pathogenesis of abdominal pain in bowel obstruction: role of mechanical stress-induced upregulation of nerve growth factor in gut smooth muscle cells

Colon distention induces persistent visceral hypersensitivity by mechanotranscription of pain mediators in colonic smooth muscle cells

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