28-day repeated oral dose toxicity study of an aqueous extract of Gryllus bimaculatus in sprague-dawley rat.

This study was performed to demonstrate a No Observed Adverse Effect Level (NOAEL) for an aqueous extract of Gryllus bimaculatus. According to other studies, using dried material or extract with ethanol or methanol determined a NOAEL dose of 1000 mg/kg or 5000 mg/kg in rats. Therefore, the Gryllus bimaculatus groups were administered orally at doses of 0, 1000, 2000, and 3000 mg/kg for four weeks. Two-week recovery groups were administered at doses of 0, and 3000 mg/kg. During administration and recovery period, the animals were observed for clinical signs, change of body weight, food consumption, hematology, and clinical chemistry. Rats in each group were periodically sacrificed, and organs were weighed and examined histologically. No difference arose between any of the dosage groups and the control group in clinical signs, histopathological examination, hematology, or clinical chemistry. In conclusion, 3000 mg/kg is a NOAEL dose for Gryllus bimaculatus extracts in Sprague Dawley rats.

Monoclonal antibody against α-actinin 4 from human umbilical vein endothelial cells inhibits endothelium-dependent vasorelaxation.

BACKGROUND: This study was attempted to identify new molecules expressed on the plasma membrane of human umbilical vein endothelial cells (HUVECs) using monoclonal antibody-based proteomics technology and to determine the effect of the identified antibody on vascular reactivity. METHODS: Twenty-two antibodies were developed from rats inoculated with HUVECs, and their effects were determined by observing vascular reactivity. RESULTS: Among the 22 antibodies, the C-7 antibody significantly inhibited endothelium-dependent vasorelaxation in response to acetylcholine (ACh) but not to histamine. Moreover, the C-7 antibody did not affect norepinephrine-induced contraction in either the endothelium-intact or -denuded aorta. A proteomics study involving immunoprecipitation of the C-7 antibody with biotinylated HUVECs showed that this antibody binds to plasma membrane proteins corresponding to immunoglobulin heavy chain (VHDJ region), chaperonin-containing T-complex polypeptide 1 and α-actinin 4. The muscarinic M3 ACh receptor and α-actinin 4 were colocalized on the plasma membrane of HUVECs, and the colocalization was found to increase in response to ACh and was inhibited by pretreatment with the C-7 antibody. CONCLUSIONS: These results demonstrate that monoclonal C-7 antibody exerts an inhibitory effect on endothelium-dependent vasorelaxation induced by ACh and that this response may at least partially result from the inhibition of α-actinin 4.