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.

Abstract White pepper (Piper nigrum L.) and long pepper (Piper longum L.) belong to family Piperaceae and are commonly used as household spices and traditional medicine worldwide, specifically in China and Southeast Asia. In Traditional Chinese Hui Medicine, these herbs are widely used for treatment of stroke. Our present study investigated effects of these herbs on inflammation in rat model with cerebral ischemia. After subjecting the rats to permanent middle cerebral artery occlusion (pMCAO) for 6 h, at doses of 100 and 200 mg/kg, dichloromethane fraction from white pepper and long pepper, respectively, was intragastrically administered once a day for seven consecutive days. Cerebral cortical and hippocampal tissues were collected after seven days. Superoxide dismutase, malonaldehyde, tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and IL-6 were measured by spectrophotometer. Phytochemical profile of dichloromethane fraction was determined through HPLC. Dichloromethane fraction exhibited anti-inflammatory activity by suppressing expression or production of IL-1β, IL-6, and TNF-α. By contrast, dichloromethane fraction showed activity against pMCAO injury by reducing oxygen-free radicals through increased superoxide dismutase activity and decreased malonaldehyde level. HPLC analysis revealed piperine as major component of dichloromethane fraction. These results show that dichloromethane fraction provides protection against cerebral ischemia. The possible mechanism is related to anti-inflammatory activity and reduction in oxygen-free radicals.

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.

Abstract To study the hepatoprotective effect of the essential oil of Artemisia capillaris Thunb., Asteraceae, on CCl4-induced liver injury in mice, the levels of serum aspartate aminotransferase and alanine aminotransferase, hepatic levels of reduced glutathione, activity of glutathione peroxidase, and the activities of superoxide dismutase and malondialdehyde were assayed. Administration of the essential oil of A. capillaris at 100 and 50 mg/kg to mice prior to CCl4 injection was shown to confer stronger in vivo protective effects and could observably antagonize the CCl4-induced increase in the serum alanine aminotransferase and aspartate aminotransferase activities and malondialdehyde levels as well as prevent CCl4-induced decrease in the antioxidant superoxide dismutase activity, glutathione level and glutathione peroxidase activity (p < 0.01). The oil mainly contained β-citronellol, 1,8-cineole, camphor, linalool, α-pinene, β-pinene, thymol and myrcene. This finding demonstrates that the essential oil of A. capillaris can protect hepatic function against CCl4-induced liver injury in mice.

Aconitum flavum Hand.-Mazz., Ranunculaceae, has been used for the treatment of rheumatism, traumatic injury in folk and clinical medicine, but the alkaloids has high toxicity. This study was designed to investigate the acute toxicity, anti-inflammatory and antinociceptive activities of non-alkaloids fractions from A. flavum in rodents. The anti-inflammatory activity was evaluated by inflammatory models of dimethylbenzene-induced ear vasodilatation and acetic acid-induced capillary permeability enhancement test in mice and carrageenan-induced paw edema in rats whereas the antinociceptive activity was evaluated using acetic acid-induced writhes, hot plate test and formalin test in mice. The result showed that the LD50 value of BtOH and EtOAc fractions could not be determined as no lethality was observed up to 40 g/kg (p.o.) in mice. BtOH fraction significantly decreased the dimethylbenzene-induced ear vasodilatation, carrageenan-induced paw edema and acetic acid-induced capillary permeability. EtOAc fraction only significantly attenuated paw edema and capillary permeability at the dose of 500 mg/kg. In antinociceptive test, BtOH and EtOAc fractions significantly reduced the writhing number evoked by acetic acid injection and the licking time in both phases of the formalin test. Meanwhile BtOH and EtOAc fractions had significant effect on hot plate test after 90 min. Our data indicate that the BtOH and EtOAc fractions of NAF are no toxicity. BtOH and EtOAc fractions not only inhibit inflammatory and peripheral inflammatory pain but also have central antinociceptive effect.