Anti-inflammatory effect of glycosaminoglycan derived from Gryllus bimaculatus (a type of cricket, insect) on adjuvant-treated chronic arthritis rat model.

Anti-inflammatory effects of glycosaminoglycan (GAG) derived from cricket (Gryllus bimaculatus, Gb) were investigated in a complete Freund's adjuvant (CFA)-treated chronic arthritic rat model. This GAG produced a significant anti-edema effect as evidenced by inhibition of C-reactive protein (CRP) and rheumatoid factor, and interfered with atherogenesis by reducing proinflammatory cytokine levels of (1) vascular endothelial growth factor (VEGF) production in human umbilical vein endothelial cells (HUVEC), (2) interleukin-6, (3) prostaglandin E2-stimulated lipopolysaccharide in RAW 264.7 cells, and (4) tumor necrosis factor (TNF)-α production in normal splenocytes, in a dose-dependent manner. This GAG was also found to induce nitric oxide (NO) production in HUVEC cells and elevated endothelial nitric oxide synthase (eNOS) activity levels. Histological findings demonstrated the fifth lumbar vertebrae (LV) dorsal root ganglion, which was linked to the paw treated with Gb GAG, was repaired against CFA-induced cartilage destruction. Further, combined indomethacin (5 mg/kg)-Gb GAG (10 mg/kg) inhibited more effectively CFA-induced paw edema at 3 h and 2 or 3 d after treatment to levels comparable to only the anti-inflammatory drug indomethacin. Ultraviolet (UV)-irritated skin inflammation also downregulated nuclear factor κB (NFκB) activity in transfected HaCaT cells. Data suggest that the anti-inflammatory effects of GAG obtained from cricket (Gb) may be useful for treatment of inflammatory diseases including chronic arthritis.

Immune Modulation of B. terrestris Worker (a Type of Bumblebee), Extract on CFA-induced Paw Edema in Rats.

To develop a composition for enhancing immunity, based on alcohol extracts of the bumblebee as an active ingredient, bumblebee ethanol extracts were evaluated for their protective effect in chronic models of inflammation, adjuvant induced rat arthritis. B. terrestris worker extract (SDIEX) and, B. hypocrita sapporoensis lava an pupa extract (SPDYBEX), significantly decreased paw edema in arthritic rats, at a dose 100 mg/kg, respectively. The cytokine levels related inflammation of COX-2, sPLA2, VEGF, and TNF-α, were decreased, compared to positive control, indomethacin (5 mg/kg). Histopathological data demonstrated decreases inflammatory activity, hind paw edema, and repaired hyaline articular cartilage in DRG over a 2 wk administration. HPLC and GC-MS analysis of SDIEX and SPDYBEX revealed the presence of cantharidin.

Blood-pressure-lowering effect of glycosaminoglycan derived from Isaria sinclairii in spontaneously hypertensive rats.

The antihypertensive effects of both extracts and glycosaminoglycan derived from Isaria sinclairii (IS) were investigated in a spontaneously hypertensive rat (SHR) model. Groups of rats were treated orally with 30 mg/kg each of: (1) saline control or extracts of (2) water-IS (3) methanol-IS, (4) butanol-IS, (5) ethyl acetate-IS, or (6) captopril as positive control. The 30-mg/kg dose was administered with a standard diet every day for a period of 2 wk. The antihypertensive effects of the individual extracts were in the following order: methanol > water > ethyl acetate > butanol. Glycosaminoglycan (GAG) obtained from IS as a water-soluble alcohol precipitation fraction produced an antihypertensive effect. One month following administration of GAG derived from IS to SHR animals there was a marked decrease in systolic blood pressure from 183 to 105 mm Hg and reduced diastolic blood pressure from 148 to 80 mm Hg compared to untreated control SHR rats. It was found that GAG produced an antihypertensive effect, which was more effective than the positive control captopril. In the SHR animal model a fall of 19% in body weight was observed in the group that received GAG. Data thus indicate that GAG derived from I. sinclairii may be a potent, naturally occurring antihypertensive agent.

Anti-inflammatory Effect of Bumblebee Alcohol Extracts in CFA-Induced Rat Edema.

In this study, we prepared alcohol extracts of the larva, pupa, queen, and cocoon (clony) of B. ignitus, B. terrestris, and B. h. sapporoensis, and tested the anti-inflammatory activity of the extracts by using a rat model of adjuvant-induced edema. The extracts derived from the queen of B. ignitus, the queen of B. terrestris, and the cocoon of B. ignitus decreased hind paw edema after 1 day of i.p. administration. These extracts also induced vasorelaxation and NO production in calf pulmonary artery endothelial cells. These results suggest that bumblebee alcohol extracts has anti-inflammatory and vasorelaxant properties.

Thirteen-Week Oral Dose Toxicity Study of G. bimaculatus in Sprague-Dawley Rats.

Gryllus bimaculatus (Gb) was orally administered at doses of 0, 0.04, 0.2, 1 and 5 g/kg bw/day for 13 consecutive weeks. There were no observed clinical signs or deaths related to treatment in all the groups tested. Therefore, the approximate lethal oral dose of G. bimaculatus was considered to be higher than 5 g/kg in rats. Throughout the administration period, no significant changes in diet consumption, ophthalmologic findings, organ weight, clinical pathology (hematology, clinical chemistry, coagulation, and urinalysis) or gross pathology were detected. Minor changes were found in hematological parameters for the 5 g/kg Gb-treated group (triglyceride reduction of 35.8%), but all changes were within normal physiological ranges. Microscopic examination did not identify any treatment-related histopathologic changes in the organs of Gb-treated rats in the high dose group. From these results, one can conclude that the no-observed adverse effect level (NOAEL) of G. bimaculatus is higher than 5 g/kg bw/day in rats.

Fatty Acid Composition of Adipose Tissues in Obese Mice and SD Rats Fed with Isaria sinclairii Powder.

Isaria sinclairii (Cicada Dongchunghacho) was studied as a potential crude natural food in powdered form. The role of tissue fatty acids in relation to the anti-obesity effects of I. sinclairii (IS) was examined by feeding the powder to SD rats ad libitum at 0, 1.25, 2.5, 5 and 10% (calculated about 8 g/kg) of the feed for a period of 3 months and 6 months. The fatty acid composition profile as indicated GC-MS, showed significantly slight dose-dependent increases in the levels of unsaturated fatty acids, particularly, arachidonic acid (C20: 4n6) , oleic acid, linoleic acid, eicosadienoic acid, eicosapentaenoic acid (EPA) (C20: 5) concentration in the the ad libitum IS-fed groups compared to the control group in SD abdominal fat over 6 month period. Over viewing of the SD and Ob mice treated Isaria sinclairii powder; there were increases in the single (mono) unsaturated fatty acids ratio but decreases in polyunsaturated fatty acid. In IS-fed groups in proportion to the treatment period, this Dongchunghacho also induced an increase in the level of same result of unsaturated fatty acid in C57BL/6 obese (ob/ob) mice over a 6-month period treatment compared to those given 10% dry mulberry leaf powder (ML) or silkworm powder mixed with the standard diet.

Single Oral Dose Toxicity Study of an Alcohol Extract of Bumblebee, Bombus ignitus Larvae in Rats.

The alcohol extract of the larvae of Bombus ignitus, otherwise known as the Bumblebee, was orally administered to rats at doses of 0, 0.04, 0.2, 1 or 2 g/kg as a single oral dose. There were no observed clinical signs or deaths related to treatment in all the groups tested. Therefore, the approximate lethal dose of the alcohol extract of B. ignitus was considered to be higher than 2 g/kg in rats. Mild decreases in body weight gain in male rats were observed dose-dependently within the B. ignitus treated groups over 2 weeks. Throughout the administration periods, no significant changes in diet consumption, ophthalmologic findings, clinical pathology (hematology, clinical chemistry and coagulation) or gross pathology were detected. Minor changes in male rats were found with in the hematological parameters in groups treated with the 0.04 g/kg, 1 g/kg or 2 g/kg of B. ignitus larvae extract, however, all the changes observed were within the physiological range. From these results, it was concluded that there was no evidence of specific toxicity related to the ingestion of alcohol extract of B. ignitus larvae.

Elevation of Nitric Oxide Synthase Activity by Dimethyladenosine from Silkworm Pupae in Aged Rats.

This study examined the mechanisms underlying the effects of the vasorelaxation active substance (VAS), dimethyladenosine-5'-L-arabinose, and its partial purification fraction on nitric oxide synthase in improving erectile dysfunction with particular focus on the nitric oxide (NO)-cGMP pathways. Two rat models, 9-month-old SD rats and 11-month-old SD rats, were given VAS (40 mg/kg per day) for 4 days, The aqueous fraction of silworm male pupae extract; semi-purified VAS (100 mg/kg per day) for 10 days, respectively. The NOS activities of the following three enzymes were examined: neuronal NO synthase (nNOS), inducible NOS (iNOS), endothelial NOS (eNOS), vascular endothelial growth factor on endothelial cells (VEGF) and anti-inflammation effect of Tumor necrosis factor-α. The results showed increases in the nitric oxide synthase activities. Western blotting of the tissue homogenate showed an increase in the nNOS level in the brain and tongue, and an increase in the endothelial NO synthase (eNOS) level in penis. However, there was little association with VEGF production in HUVEC endothelial cells and no relationship with TNF-α which showed low levels.