Evidence for up-regulation of purinergic receptor genes associating with TRPV1 receptors and neurotrophic factors in the inflamed human esophagus.

Purinergic receptors are implicated in nociceptive signaling in small primary afferents via activation of adenosine triphosphate (ATP). ATP appears to mediate HCl-induced transient receptor potential vanilloid receptor 1 (TRPV1) activation in esophageal mucosa. Up-regulation of TRPV1 expression in gastroesophageal reflux disease (GERD) is associated with increased nerve growth factor (NGF) and glial derived neurotrophic factor (GDNF). This study aims to genetically determine the expression of purinergic receptors in severe inflamed human esophagus. Distal esophageal biopsies from the subjects with erosive GERD, asymptomatic patients (AP) and healthy ones were examined. Using real-time qPCR for detecting purinergic receptors (P2X2, P2X3, P2X7, P2Y1, P2Y2, P2Y4, P2Y6 and P2Y12), TRPV1, TRPV4, NGF, and GDNF was done in this study. Both P2X3 and P2X7 mRNA expressions in GERD patients significantly increased than those in healthy controls (P < 0.001) and AP (P < 0.001), but P2X2, P2Y1, P2Y2, P2Y4, P2Y6, P2Y12 or P2Y12 had no difference within the control, AP or GERD subjects. The well correlated expression in P2X3 gene with TRPV1 (r = 0.46, P = 0.002), NGF (r = 0.54, P = 0.0002), and GDNF (r = 0.64, P = 0.0001) was found. The P2X7 gene expressions also well correlated with TRPV1 (r = 0.47, P = 0.002), NGF (r = 0.32, P = 0.037), and GDNF (r = 0.42, P = 0.005). These results suggest that chronic esophagitis increases mRNA expressions of P2X3 and P2X7 receptors accompanied by up-regulation of TRPV1 and neurotrophic factors (NGF and GDNF). These genetical alterations in esophageal mucosa might mediate sensitization of inflamed human esophagus.

Evidence for neurotrophic factors associating with TRPV1 gene expression in the inflamed human esophagus.

BACKGROUND: Transient receptor potential vanilloid-1 (TRPV1) receptor has been implicated in the mechanism of acid induced inflammation in gastro-esophageal reflux disease (GERD). It has been demonstrated that the increase in nerve growth factor (NGF) and glial derived neurotrophic factor (GDNF) was associated with the increased expression of TRPV1. We aimed to determine whether expression of TRPV1 was increased in severe inflamed human esophagus, and to test the hypothesis whether the expression of TRPV1 was mediated by neurotrophic factors such as NGF and GDNF. METHODS: We compared biopsies taken from the distal esophagus of 15 patients with erosive GERD, 16 asymptomatic patients (AP), and 10 healthy controls. We assessed the biopsies with reverse transcription polymerase chain reaction (RT-PCR) and real-time quantitative polymerase chain reaction (qPCR) for TRPV1, NGF, and GDNF. Immunohistochemical analysis of TRPV1 protein expression was also determined. KEY RESULTS: Transient receptor potential vanilloid-1 mRNA level and its protein expression were significantly greater in patients with erosive esophagitis than AP (P < 0.001) and healthy controls (P < 0.001). Nerve growth factor and GDNF gene levels in the esophageal mucosa were also significantly increased in patients with erosive esophagitis compared with AP and healthy controls (all P < 0.001). Transient receptor potential vanilloid-1 mRNA correlated well with NGF (r = 0.61, P < 0.001) and GDNF (r = 0.58, P < 0.001). CONCLUSIONS & INFERENCES: These results support the association of NGF and GDNF in the up-regulation of TRPV1 receptors in patients with erosive esophagitis.

Abnormal expressions of circadian-clock and circadian clock-controlled genes in the livers and kidneys of long-term, high-fat-diet-treated mice.

OBJECTIVES: Physiological and behavioral circadian rhythmicities are exhibited by all mammals and are generated by intracellular levels of circadian oscillators, which are composed of transcriptional/translational feedback loops involving a set of circadian-clock genes, such as Clock, Per1-3, Cry1-2, Bmal1, Dbp, E4BP4 and CK1varepsilon. These circadian-clock genes play important roles in regulating circadian rhythms and also energy homeostasis and metabolism. Determining whether obesity induced by high-fat diet affected the expressions of circadian-clock genes and their related genes in peripheral tissues, was the main focus of this study. To address this issue, we fed male C57BL/6 mice a high-fat diet for 11 months to induce obesity, hyperglycemic, hypercholesterolemic and hyperinsulinemic symptoms, and used quantitative real-time reverse transcription-PCR to measure gene expression levels. RESULTS: We found that the expressions of circadian-clock genes and circadian clock-controlled genes, including Per1-3, Cry1-2, Bmal1, Dbp, E4BP4, CK1varepsilon, PEPCK, PDK4 and NHE3, were altered in the livers and/or kidneys. CONCLUSIONS: These results indicate that obesity induced by high-fat diet alters the circadian-clock system, and obesity and metabolic syndrome are highly correlated with the expressions of circadian-clock genes and their downstream, circadian clock-controlled genes.