Optimization for extraction of an oil recipe consisting of white pepper, long pepper, cinnamon, saffron, and myrrh by supercritical carbon dioxide and the protective effects against oxygen-glucose deprivation in PC12 cells

ABSTRACT This study is to investigate the most efficient extractives of extracting oil recipe for stroke treatment and the protective effects on an oxygen and glucose deprivation model in PC12 cells. An orthogonal experimental design L9 (34) was carried out for oil recipe's optimization with supercritical CO2 fluid extraction. 2-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and enzyme-linked immunosorbent assay were conducted to evaluate cell activity and indexes in the cell lysate. The result showed that the optimum extraction condition was 30 Mpa, 50 ºC, 100 min, the extracts were analyzed by gas chromatography-mass spectrometry and among forty detected compounds 27 were identified, representing 80.86% of the total oil content. trans-Cinnamaldehyde (14.14%), piperine (9.32%), β-amyrin (6.79%), lupenone (6.28%), longifolene (6.07%), β-caryophyllene (5.21%), α-bisabolol (4.11%), and β-bisabolene (2.56%) were high mass fraction. Oil recipe could significantly attenuate PC12 cell damage, the lactate dehydrogenase release and decreased the malondialdehyde levels, glutathione peroxidase and nicotinamide adenine dinucleotide phosphate oxidase activity, glutathione and nitric oxide content (p < 0.01) and increased the level of superoxide dismutase after oxygen and glucose deprivation. The protective mechanism may be related to oil recipe's antioxidant effect by scavenging free radicals.

A subchronic toxicity study of ethanol root extract of baked Aconitum flavum in rats

ABSTRACT The genus Aconitum has strong toxicity, but the acute toxicity of baked Aconitum flavum Hand.-Mazz., Ranunculaceae, was reduced significantly on the premise of keeping anti-inflammatory and anti-nociceptive activities. However, the risk associated with long-term use is unknown. In a sub-chronic toxicity study, rats were orally administered A. flavum at doses of 0.76–3.03 g/kg for 90 days and further recovered for 14 days. Our results showed that oral treatment with A. flavum for 90 days caused significant changes in some hematological indicators at doses of 3.03 and 1.52 g/kg, such as red blood cell, hemoglobin, mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration. These results indicated that the A. flavum affects the structure and function of red blood cell. Furthermore, significant changes were observed in the white blood cell at dose of 3.03 g/kg in male rats, which confirmed tissue damage or toxicity. The liver function tests exhibited non-significant alterations in aspertate aminotransferase, alanine aminotransferase and avenin-like storage proteinsgene. But other parameters, such as total protein and albumin were obviously decreased at all doses. A. flavum also caused a significant decrease in glucose, cholesterol and triacylglyceride at all doses. For kidney function, there were significant elevations in urea and creatinine at doses of 3.03 and 1.52 g/kg. The levels of certain electrolytes (Na+, K+ and Cl-) were significantly different after 90 days of treatment with A. flavum (3.03 and 1.52 g/kg). Organs were observed by light microscopy after hematoxylin-eosin staining. Hemosiderin depositions in the spleen were observed in the A. flavum group. These data demonstrated that the subtoxicity of A. flavum was reduced considerably by baked, but the subchronic toxicity effects on the liver, kidney and spleen should not be ignored.