• Natural Product Sciences
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• v.6 no.1
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• pp.20-24
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• 2000

The enantiomers of higenamine were directly separated by high performance liquid chromatography using a chiral stationary phase and detected by UV. The R- and S-isomers of higenamine were eluted at the retention time of 22 min and 27 min, respectively. Higenamine was determined to be present as R-(+)-enantiomer not only in the embryo of Nelumbo nucifera, from which the separation of R-(+)-higenamine was reported, but also in various Aconite roots, from which higenamine was separated as optically inert racemic mixtures.

• Natural Product Sciences
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• v.5 no.4
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• pp.181-185
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• 1999

Higenamine is a cardiotonic constituent of Aconite root, one of the most important oriental traditional medicine. Since Aconite root contains toxic aconitine alkaloids, variously processed roots have been often used. Much works have been done with the chemical significances concerning with the toxic aconitine alkaloids during the processing periods. However, effects of processing on higenamine have not yet been previously studied. In this paper, the extract pattern and the amounts of higenamine extracted with water from unprocessed and processed Aconite roots were compared. R-(+)-isomer was the only higenamine enantiomer detected although racemic higenamine was reported to be separated from Aconitum spp. Sonication for 1 hour resulted in higher higenamine extraction $(12.3\;{\mu}g/g)$ than boiling water extraction for 3 hours $(6.7\;{\mu}g/g)$ of unprocessed Aconite root. Extraction of not only higenamine but also most of the other components of unprocessed Aconite roots were reduced with boiling in water. Similarly, reduced extraction was observed with extracts of all three processed Aconite roots (Kyung-Po-Aconite root, Dang-Po-Aconite root and Huk-Peon-Aconite root) by either sonicated extraction or boiling water extraction.

• Plant Resources
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• v.6 no.1
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• pp.81-88
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• 2003

Higenamine [1-(4-hydroxy-6, 7-dihydroxy-l, 2, 3, 4-tetrahydroisoquinoline) is a cardiotonic constituent of Aconiti tuber, one of the most widely prescribed oriental medicines. S-(-)higenamine was reported to have a stronger cardiotonic activity than R-(+)-higenamine and known as a central intermediate in the biosynthesis of various benzyl isoquionoline alkaloids in plants. The separation of higenamine enantiomers has been accomplished with capillary electrophoresis using cyclodextrins (CDs) as chiral selectors. Good resolution of this enantiomers was obtained using a 50 mM sodium phosphate buffer containing hydroxypropyl $\beta$-CDs using 27 cm fused silica capillary (50${\mu}{\textrm}{m}$ i.d., 20 cm to detector) at 25 $^{\circ}C$. With the electric field of 340 V/cm, the separation time of higenamine enantiomers was less than 6 min. Under this optimum conditions, the relative standard deviations of migration time and peak area were less than 1.6% and 3.2%. A 512-channel diode array detector was confirmed for the higenamine. The detection limits (S/N = 3) of these enantiomers are $1.5mutextrm{m}$/mL. We confirmed the chiral form of higenamine in medicinal plants.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2004.04a
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• pp.3-10
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• 2004

Oxidative stress is a constant threat to all living organisms and an immense repertoire of cellular defense systems is being employed by most pro- and eukaryotic systems to eliminate or to attenuate oxidative stress. Ischemia and reperfusion is characterized by both a significant oxidative stress and characteristic changes in the antioxidant defense. Heme oxigenase-l (HO-l) is up-regulated by various stimuli including oxidative stress so that it is thought to participate in general cellular defense mechanisms against ischemic injury in mammalian cells. Higenamine, an active ingredient of Aconite tuber, has been shown to have antioxidant activity along with inhibitory action of inducible nitric oxide synthase (iNOS) expression in various cells. In the present study, we investigated whether higenamine and related analogs protect cells from oxidative cellular injuries by modulating antioxidant enzymes, such as HO-l, MnSOD etc. R-form of YS-51 was the most potent inducer of HO-l in bovine endothelial cells, which inhibited apoptotic cell death by H$_2$O$_2$. HO-1 induction by YS 51 was mediated by PI3 kinase activation in which PKA- as well as PKG pathway is considered as important regulators. YS-51 also induced Mn-SOD mRNA expression by activating c-jun N-terminal kinase in endothelial cells and Hela cells. In ROS 17/2.1 cells, higenamine and enetiomers of related compounds inhibited iNOS expression by cytokine mixtures. Taken together, higenamine and related compounds can be developed as possible protective agents from oxidative cell injury or death.

• Biomolecules & Therapeutics
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• v.10 no.2
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• pp.71-77
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• 2002

Heme oxygenase-1 (HO-1) is the inducible from of the rate-limiting enzyme of heme degradation; it regulates the cellular contents of heme. HO-1 is up-regulated by various stimuli including oxidative stress so that it is thought to participate in general cellular defense mechanisms against oxidative stress in mammalian cells. To investigate the role of the cAMP-dependent protein kinase A (PKA) signaling pathway on nitrogen oxidative stress-induced HO-1 gene expression, RAW 264.7 cell cultures were treated with sodium nitroprusside (SNP). SNP increased the expression of HO-1 mRNA and protein, time- and concentration-dependently. Treatment with H89, PKA inhibitor, but not LY83583, guanylate cyclase inhibitor, significantly diminished the HO-1 expression by SNP, indicating that cAMP plays a crucial role in the induction of HO-1. Incubation with cAMP-elevating agents, such as forskolin or isoproterenol resulted in up-regulation of the expression of HO-1. Forskolin-induced expression of HO-1 was inhibited by H89. Furthermore, propranolol, $\beta$-adrenoceptor blocker, inhibited the isoproterenol-induced HO-1 expression, supporting the importance of cAMP in the induction of HO-1 expression. Higenamine-S, but not higenamineR, enhanced the HO-1 expression induced by SNP. Furthermore, cellular toxicity induced by hydrogen peroxide was attenuated by the presence of SNP, which was further increased by the presence of ZnPPIX, HO-1 inhibitor. Collectively, these results strongly suggest that up-regulation of HO-1 expression in RAW 264.7 cells involves PKA signal pathway.