Desired and side effects of the supplementation with l-glutamine and l-glutathione in enteric glia of diabetic rats.

BACKGROUND/AIMS: Enteric neuropathy associated with Diabetes Mellitus causes dysfunction in the digestive system, such as: nausea, diarrhea, constipation, vomiting, among others. The aim of this study was to compare the effects of supplementation with 2% l-glutamine and 1% l-glutathione on neurons and enteric glial cells of ileum of diabetic rats. METHODS: Thirty male Wistar rats have been used according to these group distributions: Normoglycemic (N), Normoglycemic supplemented with l-glutamine (NG), Normoglycemic supplemented with l-glutathione (NGO), Diabetic (D), Diabetic supplemented with l-glutamine (DG) and Diabetic supplemented with l-glutathione (DGO). After 120days, the ileum was processed for immunohistochemistry of HuC/D and S100ß. Quantitative and morphometric analysis have been performed. RESULTS: Diabetic rats presented a decrease in the number of neurons when compared to normoglycemic animals. However, diabetes was not associated with a change in glial density. l-Glutathione prevented the neuronal death in diabetic rats. l-Glutathione increased a glial proliferation in diabetic rats. The neuronal area in diabetic rats increased in relation to the normoglycemics. The diabetic rats supplemented with l-glutamine and l-glutathione showed a smaller neuronal area in comparison to diabetic group. The glial cell area was a decreased in the diabetics. The diabetic rats supplemented with l-glutamine and l-glutathione did not have significant difference in the glial cell body area when compared to diabetic rats. CONCLUSION: It is concluded that the usage of l-glutamine and l-glutathione as supplements presents both desired and side effects that are different for the same substance in considering normoglycemic or diabetic animals.

Myosin Va but Not nNOSα is Significantly Reduced in Jejunal Musculomotor Nerve Terminals in Diabetes Mellitus.

Nitric oxide (NO) mediated slow inhibitory junction potential and mechanical relaxation after electrical field stimulation (EFS) is impaired in diabetes mellitus. Externally added NO donor restore nitrergic function, indicating that this reduction result from diminution of NO synthesis within the pre-junctional nerve terminals. The present study aimed to investigate two specific aims that may potentially provide pathophysiological insights into diabetic nitrergic neuropathy. Specifically, alteration in nNOSα contents within jejunal nerve terminals and a local subcortical transporter myosin Va was tested 16 weeks after induction of diabetes by low dose streptozotocin (STZ) in male Wistar rats. The results show that diabetic rats, in contrast to vehicle treated animals, have: (a) nearly absent myosin Va expression in nerve terminals of axons innervating smooth muscles and (b) significant decrease of myosin Va in neuronal soma of myenteric plexus. In contrast, nNOSα staining in diabetic jejunum neuromuscular strips showed near intact expression in neuronal cell bodies. The space occupancy of nitrergic nerve fibers was comparable between groups. Normal concentration of nNOSα was visualized within a majority of nitrergic terminals in diabetes, suggesting intact axonal transport of nNOSα to distant nerve terminals. These results reveal the dissociation between presences of nNOSα in the nerve terminals but deficiency of its transporter myosin Va in the jejunum of diabetic rats. This significant observation of reduced motor protein myosin Va within jejunal nerve terminals may potentially explain impairment of pre-junctional NO synthesis during EFS of diabetic gut neuromuscular strips despite presence of the nitrergic synthetic enzyme nNOSα.

L-glutamine supplementation prevents myenteric neuron loss and has gliatrophic effects in the ileum of diabetic rats.

BACKGROUND: Peripheral neuropathy caused chronically by diabetes mellitus is related to exacerbation of oxidative stress and a significant reduction in important endogenous antioxidants. L: -Glutamine is an amino acid involved in defense mechanisms and is a substrate for the formation of glutathione, the major endogenous cellular antioxidant. AIM: This study investigated the effects of 2% L: -glutamine supplementation on peripheral diabetic neuropathy and enteric glia in the ileum in rats. METHODS: Male Wistar rats were divided into four groups: normoglycemics (N), normoglycemics supplemented with L: -glutamine (NG), diabetics (D), and diabetics supplemented with L: -glutamine (DG). After 120 days, the ileums were processed for HuC/D and S100 immunohistochemistry. Quantitative and morphometric analysis was performed. RESULTS: Diabetes significantly reduced the number of HuC/D-immunoreactive myenteric neurons per unit area and per ganglion in group D compared with normoglycemic animals (group N). L: -Glutamine (2%) prevented neuronal death induced by diabetes (group DG) compared with group D. The glial density per unit area did not change with diabetes (group D) but was significantly reduced after L: -glutamine supplementation (groups NG and DG). Ganglionic glial density was similar among the four groups. The neuronal area was not altered in groups D and DG. Glial size was reduced in group D; this was reversed by L: -glutamine supplementation (group DG). CONCLUSIONS: We concluded that 2% L: -glutamine had neuroprotective effects directly on myenteric neurons and indirectly through glial cells, which had gliatrophic effects.

Immunohistochemical study of vasoactive intestinal peptide (VIP) enteric neurons in diabetic rats supplemented with L-glutamine.

The purpose of this work was to study the area of the varicosities of nerve fibers of myenteric neurons immunoreactive to vasoactive intestinal peptide (VIP-IR) and of the cell bodies of VIP-IR submucosal neurons of the jejunum of diabetic rats supplemented with 2% L-glutamine. Twenty male rats were divided into the following groups: normoglycemic (N), normoglycemic supplemented with L-glutamine (NG), diabetic (D) and diabetic supplemented with L-glutamine (DG). Whole-mounts of the muscle tunica and the submucosal layer were subjected to the immunohistochemical technique for neurotransmitter VIP identification. Morphometric analyses were carried out in 500 VIP-IR cell bodies of submucosal neurons and 2000 VIP-IR varicosities from each group. L-Glutamine supplementation to the normoglycemic animals caused an increase in the areas of the cell bodies (8.49%) and varicosities (21.3%) relative to the controls (P < 0.05). On the other hand, there was a decrease in the areas of the cell bodies (4.55%) and varicosities (28.9%) of group DG compared to those of group D (P < 0.05). It is concluded that L-glutamine supplementation was positive both to normoglycemic and diabetic animals.

Myenteric neurons and intestinal mucosa of diabetic rats after ascorbic acid supplementation.

AIM: To investigate the effect of ascorbic acid (AA) dietary supplementation on myenteric neurons and epithelial cell proliferation of the jejunum of adult rats with chronic diabetes mellitus. METHODS: Thirty rats at 90 d of age were divided into three groups: Non-diabetic, diabetic and diabetic treated with AA (DA) (1 g/L). After 120 d of treatment with AA the animals were killed. The myenteric neurons were stained for myosin-V and analyzed quantitatively in an area of 11.2 mm(2)/animal. We further measured the cellular area of 500 neurons per group. We also determined the metaphasic index (MI) of the jejunum mucosa layer of about 2500 cells in the intestinal crypts, as well as the dimensions of 30 villi and 30 crypts/animal. The data area was analyzed using the Olympus BX40 microscope. RESULTS: There was an increase of 14% in the neuronal density (792.6 +/- 46.52 vs 680.6 +/- 30.27) and 4.4% in the cellular area (303.4 +/- 5.19 vs 291.1 +/- 6.0) respectively of the diabetic group treated with AA when compared to control diabetic animals. There were no significant differences in MI parameters, villi height or crypt depths among the groups. CONCLUSION: Supplementation with AA in the diabetic animal promoted moderate neuroprotection. There was no observation of alteration of the cellular proliferation of the jejunum mucosa layer of rats with chronic diabetes mellitus with or without supplementation with AA.

Vitamin E supplementation in rats with experimental diabetes mellitus: analysis of myosin-V and nNOS immunoreactive myenteric neurons from terminal ileum.

The effect of vitamin E (1 g/kg body weight) supplementation on myosin-V and neuronal nitric oxide synthase (nNOS) immunoreactive myenteric neurons from the ileum of diabetic rats was investigated in the present study. Forty animals were divided into the following groups: normoglycemics (N), normoglycemics treated with vitamin E (NE), diabetics (D), and diabetics treated with vitamin E (DE). Quantitative and morphometric analyses were performed. The area of the tertiary plexus was also determined. Diabetes produced a 24% reduction in the number of myosin-V neurons in group D compared with group N, an effect that was accompanied by an increase in the tertiary plexus area (P < 0.05). Neuronal density was 27% higher in group NE than group N (P < 0.05). Nitrergic neuronal density was not altered as a consequence of either diabetes or vitamin E treatment. Myosin-V and nNOS immunoreactive neuronal cell body area increased significantly in group NE. The area of myosin-V and nNOS myenteric neurons also increased in group D. Vitamin E treatment (group DE) increased only the size of nitrergic neurons. The present results suggest that vitamin E elicited a neuroprotective and neurotrophic effect on the natural aging process, but with regard to diabetes, vitamin E supplementation exerted a neurotrophic effect only on nitrergic neurons.

Diabetic rats supplemented with L-glutamine: a study of immunoreactive myosin-V myenteric neurons and the proximal colonic mucosa.

We studied the neuronal density and size of myenteric neurons and the epithelial cell proliferation and crypt depth of the proximal colon in diabetic Wistar rats after supplementing them with L-glutamine (1%). The animals were divided into five groups: untreated normoglycemic (UN), L-glutamine-treated normoglycemic (NG), untreated diabetic (UD), and L-glutamine-treated diabetics 4 days (DG4) and 45 days (DG45) days after the onset of diabetes. We observed a reduction of 52.7% and 50.44% in the neuronal density of the proximal colon of the UD group compared to the UN and NG groups, respectively (P<0.05). The neuronal density found for the DG4 (32.8%) and DG45 (28.6%) groups was higher than that of the UD group (P>0.05). There were no significant differences (P>0.05) when the data relative to the area of the myenteric neuron cell bodies, metaphasic index, and crypt depth in the proximal colon were compared among experimental groups.

Effect of the ascorbic acid treatment on the NADHd-positive myenteric neurons of diabetic rats proximal colon

The aim of this work was to study the effect of the ascorbic acid on the myenteric neurons of diabetic rats proximal colon. Fifteen rats (90 days old) were divided into three groups: control, untreated diabetic and treated diabetic with ascorbic acid (DA). After 120 days of daily treatment with ascorbic acid, the intestinal segments were submitted to the NADH-diaphorase (NADHd) histochemistry technique to expose the myenteric neurons. The group DA showed a higher neuronal density (33.4 percent) when compared to the untreated diabetic animals (p < 0.05). Cellular body area of neurons was significantly larger in group DA (17.3 percent) when compared to the untreated diabetics (p < 0.05). It could be concluded that the ascorbic acid promoted a neuroprotective effect on the NADHd myenteric neurons of the proximal colon of diabetic rats.

O efeito do ácido ascórbico sobre neurônios mioentéricos do colo proximal de ratos diabéticos foi avaliado. Quinze ratos com 90 dias foram divididos nos grupos: controle, diabéticos e diabéticos tratados com ácido ascórbico (DA). Após 120 dias de tratamento diário com ácido ascórbico os segmentos intestinais foram submetidos à técnica histoquímica NADH-diaforase (NADHd). A densidade neuronal, em uma área de 14,11 mm² para cada segmento, e o perfil do corpo celular de neurônios (500 neurônios/grupo) foram avaliados. O grupo DA apresentou maior densidade neuronal (33.4 por cento) em relação aos animais diabéticos (p < 0.05). Da mesma forma, a área do corpo celular dos neurônios foi significativamente maior no grupo DA (17.3 por cento) quando comparado com o grupo diabético (p < 0.05). Concluímos que o acido ascórbico apresentou um efeito neuroprotetor sobre os neurônios mioentéricos NADHd do colo proximal de ratos diabéticos.