The endocannabinoid N-arachidonoyl dopamine (NADA) selectively induces oxidative stress-mediated cell death in hepatic stellate cells but not in hepatocytes.

The endocannabinoid system is a crucial regulator of hepatic fibrogenesis. We have previously shown that the endocannabinoid anandamide (AEA) is a lipid mediator that blocks proliferation and induces death in hepatic stellate cells (HSCs), the main fibrogenic cell type in the liver, but not in hepatocytes. However, the effects of other endocannabinoids such as N-arachidonoyl dopamine (NADA) have not yet been investigated. The NADA-synthesizing enzyme tyrosine hydroxylase was mainly expressed in sympathetic neurons in portal tracts. Its expression pattern stayed unchanged in normal or fibrotic liver. NADA dose dependently induced cell death in culture-activated primary murine or human HSCs after 2-4 h, starting from 5 µM. Despite caspase 3 cleavage, NADA-mediated cell death showed typical features of necrosis, including ATP depletion. Although the cannabinoid receptors CB1, CB2, or transient receptor potential cation channel subfamily V, member 1 were expressed in HSCs, their pharmacological or genetic blockade failed to inhibit NADA-mediated death, indicating a cannabinoid-receptor-independent mechanism. Interestingly, membrane cholesterol depletion with methyl-ß-cyclodextrin inhibited AEA- but not NADA-induced death. NADA significantly induced reactive oxygen species formation in HSCs. The antioxidant glutathione (GSH) significantly decreased NADA-induced cell death. Similar to AEA, primary hepatocytes were highly resistant against NADA-induced death. Resistance to NADA in hepatocytes was due to high levels of GSH, since GSH depletion significantly increased NADA-induced death. Moreover, high expression of the AEA-degrading enzyme fatty acid amide hydrolase (FAAH) in hepatocytes also conferred resistance towards NADA-induced death, since pharmacological or genetic FAAH inhibition significantly augmented hepatocyte death. Thus the selective induction of cell death in HSCs proposes NADA as a novel antifibrogenic mediator.

Effect of duodenal glucose and acute hyperglycemia on rectal perception and compliance in response to tension-controlled rectal distension in healthy humans.

BACKGROUND: Acute changes in blood glucose concentration affect gastrointestinal motor and sensory function. Tone and distensibility contribute to intact rectal function. AIMS: To test the effects of duodenal glucose (euglycemic hyperinsulinemia), intravenous glucose (hyperglycemic hyperinsulinemia), and saline (euglycemic normoinsulinemia as control) on rectal perception and compliance in response to tension-controlled rectal distension. METHODS: During duodenal glucose at 2 kcal min(-1), marked hyperglycemic clamp (approximately 13 mmol L(-1)), or saline as control, responses to fixed-tension rectal distension, applied by means of a computerized tensostat, were compared randomized on three separate days in eight healthy subjects. RESULTS: At discomfort level (score 3 on the 0-4 rectal score scale), perception of rectal distension was significantly higher during euglycemic hyperinsulinemia (45 +/- 3 g cm(-2) tolerance) and significantly lower during hyperglycemia (83 +/- 4 g cm(-2) tolerance), both reaching significance versus control (64 +/- 6 g cm(-2) tolerance; P < 0.05). At this level, no relevant variations of rectal compliance were seen, which were 10.3 +/- 1 mL mmHg(-1) during duodenal glucose, 9.5 +/- 1 mL mmHg(-1) for the group with hyperglycemia, and 9.7 +/- 2 mL mmHg(-1) for the control. CONCLUSION: Duodenal glucose provokes rectal hypersensitivity whereas acute hyperglycemia contributes to rectal hyposensitivity. Despite different rectal tenso-sensitivity, rectal compliance remains virtually unchanged. Any dysfunction may cause rectal complaints.

Influence of E. coli strain Nissle 1917 (EcN) on intestinal gas dynamics and abdominal sensation.

E. coli strain Nissle 1917 (EcN) is a probiotic clinically used with various indications. However, especially at the beginning of treatment, some patients report abdominal bloating. In a prospective, randomized, double-blind study in 30 healthy individuals we assessed the influences of EcN on intestinal gas dynamics and abdominal sensation. After one week without medication volunteers orally received 2.5-25 x 10(9) colony-forming units of EcN or placebo per day for 21 days. EcN was well tolerated and did not significantly affect abdominal symptoms, stool frequency or stool consistency. During gas challenge at different days no difference in the perception scores (range from 0 = no perception to 6 = pain) was observed between the two groups: the mean perception score was 1.2 (SD 0.2) in the EcN group and 1.4 (SD 0.2) in the placebo group. EcN had no relevant influence on intestinal gas dynamics.

Intestinal gas retention in patients with idiopathic slow-transit constipation.

Patients with slow-transit constipation (STC) have delayed colonic transit for solid und liquid bowel contents but intestinal gas handling has not been studied so far. Different nutrients influence motor and sensory gut function. We hypothesized that, in patients with STC, alteration of regulatory mechanisms may result in impaired intestinal gas dynamics. On 3 separate days, validated gas challenge was performed in 10 STC patients and 10 volunteers during duodenal saline, lipids, or intravenous glucose. During saline only 60% +/- 8% of gas was cleared by STC patients after 60-min gas infusion, vs. 91% +/- 2% by controls (P < 0.001). Acute hyperglycemia or lipids did not change intestinal gas dynamics in these patients (saline infusion), but compared to healthy subjects, significant intestinal gas retention occurred. In STC, disturbances of intestinal gas dynamics include basal intestinal gas retention, and this is virtually not affected by acute hyperglycemia or duodenal lipids.

Impaired intestinal gas clearance during marked hyperglycemia in patients with functional abdominal bloating.

BACKGROUND: Especially in patients with functional intestinal disorders, impaired intestinal gas transit can be involved in abdominal symptom generation. We have previously demonstrated an acceleration of intestinal gas clearance in health during acute fasting hyperglycemia and hypothesize that in patients with functional abdominal bloating this mechanism may fail. METHODS: In 14 healthy subjects and 14 patients with functional abdominal bloating we compared effects of acute fasting hyperglycemia (approximately 12 mmol/l) and during euglycemia (control studies) on intestinal gas dynamics. Gas was infused into the jejunum (12 ml/min) for 120 min while rectal gas evacuation was continuously measured; perception and abdominal girth changes were separately evaluated. RESULTS: Marked hyperglycemia accelerated gas evacuation (-98 (53) ml 1 h intestinal gas retention) in health. In patients with functional abdominal bloating, marked hyperglycemia failed to accelerate gas transit and intestinal gas retention developed (421 (116) ml 1 h intestinal gas retention, p < 0.05 vs. health) which results in increased abdominal symptoms (perception score >3) and abdominal distension (>3 mm girth increment) as compared with control subjects (p < 0.05 for both). CONCLUSION: Intestinal gas clearance is delayed in patients with functional abdominal bloating and the increase in gas clearance during acute hyperglycemia in healthy volunteers does not occur in these patients.