Minoru Kawanishi

Lowered response threshold and increased responsiveness to mechanical stimulation of cutaneous nociceptive fibers in streptozotocin-diabetic rat skin in vitro—correlates of mechanical allodynia and hyperalgesia observed in the early stage of diabetes

Rats rendered diabetic by streptozotocin (STZ) show allodynia and hyperalgesia and thus, have been offered as a model of pain in diabetic neuropathy. However, recent electrophysiological studies on these rats found that C-fiber nociceptors were not consistently hyperexcitable to mechanical stimulations by von Frey hairs and that there was no change in their response thresholds. In the present study, we used rat skin-saphenous nerve in vitro preparations, in which the receptive fields of identified single C-polymodal receptors (CPRs) can be accurately stimulated with a servo-controlled mechanical stimulator. Single fiber recordings from CPRs were performed in diabetic rats with an increased behavioral nociceptive response 7-19 days after STZ injection. The proportion of units with spontaneous activity and the magnitude of this activity increased in the diabetic preparations. The response thresholds of CPRs were significantly decreased with ramp-pressure stimulation and their response magnitude to the suprathreshold stimulation was significantly increased in diabetic rats. In addition, the response pattern to mechanical stimulation was also changed to a non-adapting type. These findings suggest that changes in CPRs contribute to the enhanced nociception observed in the early stage of diabetic neuropathy.

Involvement of oxidative stress in increases in the serum levels of various enzymes and components in rats with water-immersion restraint stress.

The plasma or serum levels of various enzymes and components are known to increase in rats with water-immersion restraint stress (WIRS). We examined whether oxidative stress is involved in increases in the serum levels of various enzymes and components in rats with WIRS. Rats were exposed to WIRS for 6 h after oral administration of vitamin E (VE) (50 or 250 mg/kg). Rats with WIRS had increased serum alanine aminotransferase, aspartate aminotranseferase, lactate dehydrogenase, creatine kinase, urea nitrogen, creatinine, glucose, corticosterone, adrenocorticotropic hormone and lipid peroxide (LPO) levels, increased kidney and heart VE levels, decreased skeletal muscle VE level, and increased LPO levels in all tissues studied. Pre-administered VE (50 or 250 mg/kg) attenuated the increased serum alanine aminotransferase, aspartate aminotranseferase, lactate dehydrogenase, creatine kinase, urea nitrogen, creatinine, and LPO levels, the decreased skeletal muscle VE level, and the increased LPO levels in all tissues studied more effectively at its higher dose than at its lower dose. However, either dose of the pre-administered VE did not affect the increased serum glucose, corticosterone, and adrenocorticotropic hormone levels. These results suggest that oxidative stress is involved in increases in the serum levels of various enzymes and components in rats with WIRS.

Vitamin E protects against stress-induced gastric mucosal lesions in rats more effectively than vitamin C.

In this study, we examined the protective effects of vitamin E (VE) against gastric mucosal lesions induced by water immersion restraint stress (WIRS) in rats in comparison with that of vitamin C (VC). The gastric mucosa of rats with 6 h of WIRS showed lesions with bleeding, decrease in nonprotein SH, VC, VE, and adherent mucus concentrations and constitutive nitric oxide synthase activity, and increase in lipid peroxide and NOx (nitrite/nitrate) concentrations and myeloperoxidase, xanthine oxidase, and inducible nitric oxide synthase activities. Either VE (0.05 or 0.5 mmol/kg) or VC (0.5 or 1.5 mmol/kg) was orally administered to rats with 6 h of WIRS just before the onset of the stress. Both doses of preadministered VE prevented gastric mucosal lesion development and attenuated all these changes in gastric mucosal components and enzymes studied, whereas only the higher dose of preadministered VC suppressed the changes in all parameters studied. These results indicate that orally administered VE protects against WIRS‐induced gastric mucosal lesions in rats more effectively than orally administered VC. These results also suggest that the administered VE protects against gastric mucosal lesions in rats with WIRS through its antioxidant and anti‐inflammatory actions in the gastric mucosa in the same way as the administered VC.

Tissue glucose level modulates the mechanical responses of cutaneous nociceptors in streptozotocin-diabetic rats but not normal rats in vitro

&NA; The maintenance of normoglycemia has been reported to reduce painful sensations in diabetic subjects. This suggests that lowering the tissue glucose concentration might inhibit the increased cutaneous nociceptor activities seen in a diabetic conditin. To test this hypothesis, we studied the effect of changing the glucose concentration in the superfusate of in vitro preparations (high, HG: 20 mM or normal glucose, NG: 6.7 mM) on the mechanical response of C‐fiber polymodal receptors (C‐polymodal receptors). Single fiber activities of C‐polymodal receptors were recorded from skin‐nerve in vitro preparations of streptozotocin‐induced diabetic and age‐matched control rats. Pressure stimulation was applied to the receptive field by a servo‐controlled mechanical stimulator. C‐polymodal receptors from diabetic preparations superfused with HG‐solution showed increased spontaneous activity, lowered response threshold, increased response magnitude and a less adaptive response pattern to mechanical stimulation compared with those from control preparations superfused with NG‐solution. C‐polymodal receptors from diabetic preparations superfused with NG‐solution showed no such changes. The responsiveness of C‐polymodal receptors from control preparations was not different in NG‐ or HG‐conditions. These data demonstrated that normalization of the glucose concentration normalized the responsiveness of C‐polymodal receptors in diabetic animals. This response may be associated with the fact that normoglycemia reduces painful sensations in diabetic subjects.

Compound 48/80 causes oxidative stress in the adrenal gland of rats through mast cell degranulation

Abstract Rats were intraperitoneally treated once with compound 48/80 (C48/80), a mast cell degranulator, (0.75 mg/kg). Serum serotonin, histamine and corticosterone levels increased 0.5 h after C48/80 treatment, but their increases were reduced thereafter. Adrenal total ascorbic acid (ascorbic acid plus dehydroascorbic acid), ascorbic acid and dehydroascorbic acid levels decreased 0.5, 3 or 6 h after C48/80 treatment, adrenal lipid peroxide level increased at 3 and 6 h, adrenal non-protein-SH level decreased at 3 and 6 h and adrenal β-tocopherol level decreased at 3 h. Ketotifen, a mast cell stabilizer (1 mg/kg) administered intraperitoneally at 0.5 h before C48/80 treatment, attenuated all these changes found in the serum and adrenal at 3 h after treatment, while β-tocopherol (250 mg/kg), administered orally at 0.5 h after C48/80 treatment, attenuated all these changes in the adrenal tissue. These results indicate that C48/80 causes oxidative stress in rat adrenal gland through mast cell degranulation.

Protective effect of L-ascorbic acid against oxidative damage in the liver of rats with water-immersion restraint stress

Abstract We examined whether L-ascorbic acid (AA) (or reduced ascorbic acid) protects against oxidative damage in the liver of rats subjected to water-immersion stress (WIRS). AA (100, 250 or 500 mg/kg) was orally administered at 0.5 h before the onset of WIRS. Rats with 6 h of WIRS had increased serum corticosterone, glucose, total ascorbic acid (T-AA), AA, lipid peroxide (LPO), and NOx concentrations and alanine aminotransferase and aspartate aminotrasferase activities. The stressed rats had increased hepatic LPO, NOx, and dehydroascorbic acid concentrations and myeloperoxidase activity, decreased hepatic T-AA, AA, reduced glutathione concentrations and superoxide dismutase activity, and unchanged hepatic vitamin E concentration. Pre-administered AA attenuated the stress-induced changes in serum LPO and NOx concentrations and alanine aminotransferase and aspartate aminotrasferase activities and hepatic LPO, NOx, and T-AA, AA, dehydroascorbic acid, and reduced glutathione concentrations and myeloperoxidase and superoxide dismutase activities dose-dependently. Pre-administered AA did not affect the stress-induced changes in serum corticosterone and glucose concentrations. These results indicate that pre-administered AA protects against oxidative damage in the liver of rats with WIRS possibly by attenuating disruption of the antioxidant defense system and increases in NO generation and neutrophil infiltration in the tissue.

Different mode of action of endothelin-1 in parietal and visceral veins

The mode of contractile effect of endothelin-1 in venous smooth muscles was evaluated using three isolated veins, the jugular vein, the portal vein and the inferior caval vein of dogs. The contractile responses to endothelin-1 were attenuated by atropine and augmented by neostigmine in the inferior caval and portal veins, but were not affected by either agent in the jugular vein. We conclude that the contractile responses of visceral veins to endothelin-1 are partly mediated via endogenously released acetylcholine.