Targeting eotaxin-1 and CCR3 receptor alleviates enteric neuropathy and colonic dysfunction in TNBS-induced colitis in guinea pigs.

BACKGROUND: The accumulation of eosinophils is mediated by the chemokine receptor-3 (CCR3)-eotaxin axis. Increased expression of eotaxin and its receptor is associated with inflammatory bowel disease (IBD). Activation of eosinophils causes the release of cationic proteins that are neurotoxic such as eosinophil-derived neurotoxin (EDN). Damage to enteric neurons alters neurally controlled functions of the gut correlated with intestinal inflammation. We hypothesized that inhibition of the CCR3-eotaxin axis will prevent inflammation-induced functional changes to the gastrointestinal tract. METHODS: Hartley guinea pigs were administered with trinitrobenzene sulfonate (TNBS; 30 mg/kg in 30% ethanol) intrarectally to induce colitis. A CCR3 receptor antagonist (SB 328437 [SB3]) was injected intraperitoneally 1 hour postinduction of colitis. Animals were euthanized 7 days post-treatment and colon tissues were collected for ex vivo studies. The EDN-positive eosinophils in the colon, indicating eosinophil activation, were quantified by immunohistochemistry. Effects of SB3 treatment on gross morphological damage, enteric neuropathy, and colonic dysmotility were determined by histology, immunohistochemistry, and organ bath experiments. KEY RESULTS: The number of EDN-positive eosinophils was significantly increased in the lamina propria in close proximity to myenteric ganglia in inflamed colon. The TNBS-induced inflammation caused significant damage to colonic architecture and inhibition of colonic motility. Treatment with SB3 antagonist attenuated inflammation-associated morphological damage in the colon, reduced infiltration of EDN-positive eosinophils and restored colonic motility to levels comparable to control and sham-treated guinea pigs. CONCLUSION & INFERENCES: This is the first study demonstrating that inhibition of CCR3-eotaxin axis alleviates enteric neuropathy and restores functional changes in the gut associated with TNBS-induced colitis.

Electrophysiological and morphological changes in colonic myenteric neurons from chemotherapy-treated patients: a pilot study.

BACKGROUND: Patients receiving anticancer chemotherapy experience a multitude of gastrointestinal side-effects. However, the causes of these symptoms are uncertain and whether these therapeutics directly affect the enteric nervous system is unknown. Our aim was to determine whether the function and morphology of myenteric neurons are altered in specimens of the colon from chemotherapy-treated patients. METHODS: Colon specimens were compared from chemotherapy-treated and non-treated patients following colorectal resections for removal of carcinoma. Intracellular electrophysiological recordings from myenteric neurons and immunohistochemistry were performed in whole mount preparations. KEY RESULTS: Myenteric S neurons from chemotherapy-treated patients were hyperexcitable; more action potentials (11.4 ± 9.4, p < 0.05) were fired in response to depolarising current pulses than in non-treated patients (1.4 ± 0.5). The rheobase and the threshold to evoke action potentials were significantly lower for neurons from chemotherapy-treated patients compared to neurons from non-treated patients (p < 0.01). Fast excitatory postsynaptic potential reversal potential was more positive in neurons from chemotherapy-treated patients (p < 0.05). An increase in the number of neurons with translocation of Hu protein from the cytoplasm to the nucleus was observed in specimens from chemotherapy-treated patients (103 ± 25 neurons/mm(2) , 37.2 ± 7.0%, n = 8) compared to non-treated (26 ± 5 neurons/mm(2) , 11.9 ± 2.7%, n = 12, p < 0.01). An increase in the soma size of neuronal nitric oxide synthase-immunoreactive neurons was also observed in these specimens. CONCLUSIONS & INFERENCES: This is the first study suggesting functional and structural changes in human myenteric neurons in specimens of colon from patients receiving anticancer chemotherapy. These changes may contribute to the causation of gastrointestinal symptoms experienced by chemotherapy-treated patients.

Human enteric neurons: morphological, electrophysiological, and neurochemical identification.

BACKGROUND: Access to tissue, difficulties with dissection, and poor visibility of enteric ganglia have hampered electrophysiological recordings of human enteric neurons. Here, we report a method to combine intracellular recording with simultaneous morphological identification of neurons in the intact myenteric plexus of human colon ex vivo. METHODS: Specimens of human colon were dissected into flat-sheet preparations with the myenteric plexus exposed. Myenteric neurons were impaled with conventional microelectrodes containing 5% 5,6-carboxyfluorescein in 20 mM Tris buffer and 1 M KCl. KEY RESULTS: Electrophysiological recordings identified myenteric neurons with S and AH type properties (n = 13, N = 7) which were dye filled and classified during the recording as Dogiel type I (n = 10), Dogiel type II (n = 2), or filamentous (n = 1) cells. This classification was confirmed after fixation, in combination with immunohistochemical characterization. CONCLUSIONS & INFERENCES: This method allows electrophysiological characterization with simultaneous identification of morphology. It can be used to identify recorded cells immediately after impalement and greatly facilitates recordings of human myenteric neurons in freshly dissected specimens of tissue. It can also be combined with immunohistochemical labeling of recorded cells.

Children born small for gestational age (SGA).

SGA (small for gestational age) is a child born with birth weight and/or length (BW/BL) under two standard deviations (2 SDS) for the gestational age and sex of the population. ~5% of all newborn children are SGA. A broad spectrum of factors are found to be causative: maternal, placental, foetal, metabolic, and genetic. In the newborn period the SGA children are at greater risk of life-threatening conditions: hypoglycaemia, hypercoagulability, necrotic enterocolitis, direct hyperbilirubinemia, hypotension, etc. Approximately 10 percent of SGA children do not achieve catch-up growth and remain short (≥-2 SDS) into adulthood. SGA people have an increased incidence of metabolic syndrome, coronary artery disease, stroke, low bone density and osteoporosis. SGA children aged more than 4 years with no evidence of spontaneous catch-up and with a height≥2.5 SD are considered for growth hormone (GH) treatment.

Effect of fish oil supplementation on aortic compliance in rats: role of the endothelium.

Arterial compliance improves with dietary fish oils in patients with high cardiovascular risk. Since fish oils alter prostaglandin metabolism and the L-arginine-nitric oxide pathway, and since compliance may be modified by vasoactive substances, the effect of the endothelium and some of its derivatives on aortic complaince were examined. Rats were randomly allocated to four groups, the first of which fed only the regular chow. The remaining three groups were fed the chow supplemented by daily gavage with either coconut, fish or safflower oil for 8 weeks. The thoracic aorta was removed and six 2 mm rings obtained. Rings were paired and one from each pair treated with either N(W)-nitro-L-arginine, indomethacin or de-endothelialized. A diameter-tension curve was initiated from wire touch position using incremental increases in wire distance until no further response observed. The data was transformed to a diameter-pressure relationship and fitted with a linear equation, the slope of which related directly to compliance. De-endothelialization (slopes: control vs de-endothelialized: 9.05+/-0.15 vs 8.31+/-0.24; P< 0.05) and indomethacin (slopes: control vs indomethacin: 9.11+/-0.15 vs 7.76+/-0.37; P< 0.05) significantly decreased arterial compliance as did dietary fish oils (slopes: control vs n-3: 9.16+/-0.11 vs 7.84+/-0.39; P< 0.05). No further effect was seen with indomethacin in the fish oil treated group. It is concluded that the endothelium and in particular, endothelium derived prostanoids, contribute to vessel compliance. We also conclude that fish oils have a similar action to indomethacin, leading to the increase in aortic stiffness observed.