Background: Dysregulation of pro-inflammatory cytokines such as tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) form the basis of immune-mediated inflammatory diseases. Vitex trifolia L. is a medicinal plant growing in countries such as China, India, Australia and Singapore. Its dried ripe fruits are documented in Traditional Chinese Medicine to treat ailments like rhinitis and dizziness. Its leaves are used traditionally to treat inflammation-related conditions like rheumatic pain. Objective: This study aimed to investigate the effects of V. trifolia leaf extracts prepared by different extraction methods (Soxhlet, ultrasonication, and maceration) in various solvents on cytokine production in human U937 macrophages, and identify phytoconstituents from the most active leaf extract. Methods: Fresh leaves of V. trifolia were extracted using Soxhlet, ultrasonication, and maceration in hexane, dichloromethane, methanol, ethanol or water. Each extract was evaluated for its effects on TNF-α and IL-1β cytokine production by enzyme-linked immunosorbent assay in lipopolysaccharide-stimulated human U937 macrophages. The most active extract was analyzed and further purified by different chemical and spectroscopic techniques. Results: Amongst 14 different leaf extracts investigated, extracts prepared by ultrasonication in dichloromethane and maceration in ethanol were most active in inhibiting TNF-α and IL-1β production in human U937 macrophages. Further purification led to the isolation of artemetin, casticin, vitexilactone and maslinic acid, and their effects on TNF-α and IL-1β production were evaluated. We report for the first time that artemetin suppressed TNF-α and IL-1β production. Gas chromatography-mass spectrometry analyses revealed the presence of eight other compounds. To the best of our knowledge, this is the first report of butylated hydroxytoluene, 2,4-di-tertbutylphenol, campesterol and maslinic acid in V. trifolia leaf extracts.
SUMMARYA respiratory drift has been demonstrated in cells isolated from petals of Aranda flowers at different stages of development. Tight buds had a respiratory quotient of 0-5; this increased to 0-7 in the first flower and reached 10 in the mature flower. There appeared to be a non-glycolytic pathway contribution in addition to the EMP pathway in tight bud and newly opened flower stages. Carbohydrate metabolism in the mature flower proceeds predominantly via the EMP pathway. Cyanide-insensitive respiration was demonstrated in the tight bud. There was a shift towards cyanide-sensitive respiration as the flower developed.
Respiration of isolated Aranda orchid petal cells increased markedly after cut flowers were treated with ethylene . An increase in respiration was observed 15 to 20 h after treatment which was further enhanced in the presence of oxygen and ethylene . Ethylene induces the development of a cyanide resistant pathway in fully opened orchid flower tissues where the cyanide resistant capacity is negligible . However, there appears a shift back to the cyanide sensitive pathway some time after induction .
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