Assessment of the analgesic, anti-inflammatory and sedative effects of the dichloromethanol extract of Schinus molle.

OBJECTIVE: Determination of the active fraction and compounds of the dichloromethanol extract of Schinus molle seeds and evaluation of their biological effects. MATERIALS AND METHODS: Dried seeds of Schinus molle were sequentially extracted in hexane, acetyl acetate and dichloromethane. The dichloromethane extract was separated into two fractions (1 and 2) by column chromatography. Fraction 2 was further separated into its two constituent compounds which were characterized as belonging to the lanosteroid group of compounds. Both factions were tested for their analgesic, anti-inflammatory and sedative effects. RESULTS: The two fractions significantly increased (p<0.05) the tail flick latency though fraction 2 provided better and more long lasting protection against thermal pain. On the other hand, the anti-inflammatory effect of ibuprofen, though inferior to the anti-inflammatory effect of fraction 2 was better than the effects of fraction 1. Fraction 2 significantly (p<0.01) reduced rat paw oedema compared to the saline treatment group throughout the experiments while fraction 2 compared to fraction 1 showed significantly (p<0.01) greater inflammatory effects. On the other hand both fractions lacked significant sedative effects. CONCLUSIONS: Given that fraction 2 had only two constituent compounds (isomasticadienonic and Masticatrienonate), one or both of these compounds should be contributing to the observed analgesic and anti-inflammatory effects.

Effect of aged garlic extract on caspase-3 activity, in vitro.

In neurodegeneration, such as Alzheimer's disease (AD), apoptosis results in the loss of valuable neurons. A key mechanism in apoptosis is the activation of caspase-3. Caspase-3 activity first becomes detectable early in apoptosis, continues to increase as cells undergo apoptosis, and rapidly declines in late stages of apoptosis. Its activity is an early marker of cells undergoing apoptosis. Caspase-3 catalyzes the formation of beta-amyloid peptide, a hallmark of AD. The purpose of the study was to determine whether dietary aged garlic extract (AGE), with known antioxidant properties and neuroprotection against Alzheimer's beta-amyloid cytotoxicity, inhibits the caspase-3 activity in vitro. Caspase-3 activity was assayed according to the prescribed protocol and incubated overnight at ambient temperature. We report that AGE inhibits caspase-3 in dose dependent manner. Caspase-8 was not inhibited by AGE. As a caspase-3 inhibitor, AGE may be effective in reducing apoptotic death of neurons since caspase inhibitors have been shown to inhibit neuronal cell death. We propose a scheme for the ameliorative effect of AGE on deleterious effects of beta-amyloid and possibly uncontrolled caspase-3 activity.

Effect of Aged Garlic Extract on the Cytotoxicity of Alzheimer ß-Amyloid Peptide in Neuronal PC12 Cells.

Alzheimer's disease (AD) is the most common cause of progressive intellectual failure and memory impairment among the elderly, affecting over 4 million Americans. A hallmark of the disease is the accumulation of ß-amyloid, whose neurotoxicity has been well documented. A therapeutic approach may be to ameliorate this toxicity. Garlic has antioxidant, anti-aging and anti-ischemic properties. The purpose of the present study was to determine whether aged garlic extract attenuates the effect of neurotoxin, ß-amyloid 25-35 (Aß) in neuronal PC12 cells. Proliferating (non-neuronal) PC12 cells were treated with the nerve growth factor for at least 4 days before experiments were initiated. This treatment differentiates PC12 cells to a neuronal cell line. Cell viability was determined using the MTS-based assay (Promega). Wakunaga of America (Mission Viejo, CA) kindly donated aged garlic extract(TM) (AGE(TM)) as Kyolic(R) liquid. AGE(TM) was sterilized by filtration and diluted in cell growth medium. Our results show that AGE™ protects neuronal PC12 cells against Aß (95nM) toxicity in a dose dependent manner. AGE(TM) may be effective in ameliorating Aß -neurotoxicity.

Neurobehavioral effects of delta 9-THC and cannabinoid (CB1) receptor gene expression in mice.

The differential sensitivity following the administration of delta 9-THC to 3 mouse strains, C57BL/6, DBA/2 and ICR mice, indicated that some of the neurobehavioral changes may be attributable to genetic differences. The objective of this study was to determine the extent to which the cannabinoid (CB1) receptor is involved in the observed behavioral changes following delta 9-THC administration. This objective was addressed by experiments using: (1) DNA-PCR and reverse PCR; (2) systemic administration of delta 9-THC, and; (3) intracerebral microinjection of delta 9-THC. The site specificity of action of delta 9-THC in the brain was determined using stereotaxic surgical approaches. The intracerebral microinjection of delta 9-THC into the nucleus accumbens was found to induce catalepsy, while injection of delta 9-THC into the central nucleus of amygdala resulted in the production of an anxiogenic-like response. Although the DNA-PCR data indicated that the CB1 gene appeared to be identical and intronless in all 3 mouse strains, the reverse PCR data showed two additional distinct CB1 mRNAs in the C57BL/6 mouse which also differed in pain sensitivity and rectal temperature changes following the administration of delta 9-THC. It is suggested that the diverse neurobehavioral alterations induced by delta 9-THC may not be mediated solely by the CB1 receptors in the brain and that the CB1 genes may not be uniform in the mouse strains.

Vitamin E and the inhibition of platelet lipoxygenase.

Lipoxygenase was assayed by the formation of hydroxy-eicosatetraenoic acid (HETE) from arachidonic acid in human and rabbit washed platelets. Platelets from vitamin E-supplemented rabbits had much less lipoxygenase activity than platelets from either vitamin E-deficient or normal rabbits. Human and rabbit platelets preincubated with vitamin E had lowered lipoxygenase activity. These data show that vitamin E inhibits platelet lipoxygenase. Vitamin E and vitamin E acetate, in vitro, were equally effective inhibitors of lipoxygenase. Tween 20, in vitro, was a highly effective inihbitor of lipoxygenase. These data show that vitamin E functions, in vitro, as a surfactant in the inhibition of platelet lipoxygenase.

Differential inhibitory effects of vitamin E and other antioxidants on prostaglandin synthetase, platelet aggregation and lipoxidase.

Prostaglandin biosynthesis from eicosa-8,11,14-trienoic acid in microsomes from the bovine vesicular gland is inhibited by the antioxidants alpha-naphthol. guaiacol, NDGA and propyl gallate. Prostaglandin biosynthesis in this system is not inhibited by the antioxidants BHT, DL-alpha-tocopherol and Trolox C. Arachidonic acid induced platelet aggregation is inhibited by specifically by alpha-naphthol. guaiacol, NDGA and propyl gallate. Both arachidonic acid induced platelet aggregation and ADP induced platelet aggregation are inhibited non-specifically by the antioxidants BHT, DL-alpha-tocopherol and Trolox C. All antioxidants tested in this study inhibit soybean lipoxidase. Thus alpha-naphthol, NDGA and propyl gallate are non-specific inhibitors of both prostaglandin synthetase and soybean lipoxidase while BHT, DL-alpha-tocopherol and Trolox C are specific inhibitors of soybean lipoxidase alone.

Fatty acids and their prostaglandin derivatives: inhibitors of proliferation in aortic smooth muscle cells.

Prostaglandins are synthesized from eicosa-8,11,14-trienoic acid and eicosa-5,8,11-14-tetraenoic acid by smooth muscle cell cultures from guinea pig aorta. Production is inhibited by indomethacin. The precursor fatty acids and their prostaglandin derivatives inhibit proliferation of the cell cultures. The relative availability of fatty acids for prostaglandin biosynthesis may represent a control mechanism for cell proliferation.