T1: Phytotherapy Research, Vol. 11, 576—582 (1997)

Cardiovascular Pharmacology of 3-n-butylphthalide in Spontaneously Hypertensive Rats

D. Tsi and B. K. H. Tan

The hypotensive and vasorelaxant effects of 3-n-butylphthalide (BuPh) and its possible mechanisms of action were investigated in spontaneously hypertensive rats (SHR) for the first time. A 13-day intraperitoneal infusion of BuPh at doses of 2.0 and 4.0 mg/day produced a transient hypotensive effect while a dose of 0.5 mg/day showed a significant hypotensive effect only on day 12. BuPh at 0.5 mg/day had no effect on the plasma and tissue angiotensin converting enzyme (ACE) activities, or on the tissue lipid peroxidation index. BuPh relaxed endothelium-intact and denuded aortic rings precontracted with phenylephrine and KCl. N G -nitro-L-arginine methyl ester, an inhibitor of nitric oxide synthase, did not attenuate the vasorelaxant activity of BuPh. The cumulative concentration response curves of phenylephrine and Ca 2+ (in CaCl2 -free, high KCl medium) were non-competitively inhibited by BuPh. However, BuPh did not interfere with the caffeine-induced release of intracellular Ca 2+ . It appears that the vasorelaxant effect of BuPh could be attributed to the blockade of Ca 2+ entry, possibly through voltage- and receptor-operated Ca 2+ channels, thereby lowering the systolic blood pressure of SHR.

2: Acta Pharmacol Sin 2000 May;21(5):433-8

Inhibitory effects of chiral 3-n-butylphthalide on inflammation following focal ischemic brain injury in rats.

Xu HL, Feng YP.

AIM: To evaluate the degree of neutrophil infiltration into ischemic tissue after transient focal cerebral ischemia, and to examine the effects of chiral 3-n-butylphthalide (NBP) on this inflammatory process. METHODS: After a 24-h reperfusion following transient cerebral ischemia, two different techniques, histologic analysis and modified myeloperoxidase (MPO)-quantification method, were utilized to identify the infiltration of neutrophils into cerebral tissue following ischemia. The expression of intercellular adhesion molecule-1 (ICAM-1) and tumor necrosis factor-alpha(TNF-alpha) in the ischemic zone were observed by immunohistochemistry, Western blot, and in situ hybridization techniques.

RESULTS: In cerebral cortex area perfused by middle cerebral artery (MCA), MPO activity was greatly increased after 24 h of reperfusion in the vehicle group, and it correlated well with the infiltration of neutrophils. Administration of dl-, d-, and l-NBP (20 mg.kg-1) partially inhibited both the increase in MPO activity and the appearance of neutrophils in ischemia-reperfusion sites. Up-regulation of ICAM-1 was also observed on the microvessel endothelium in the ischemic territory. In addition, chiral NBP markedly blunted ICAM-1 expression, and decreased the number of TNF-alpha blue purple-positive neurons induced by ischemia-reperfusion injury. CONCLUSION: The results indicate that the increase in neutrophils infiltration into the infarct site implicated postischemic brain injury, and NBP was effective in protecting the ischemic sites following ischemic insult.

PMID: 11324442 [PubMed - indexed for MEDLINE]

3: Yakugaku Zasshi 1989 Jun;109(6):402-6

[Centrally acting muscle relaxant effect of phthalides (ligustilide, cnidilideand senkyunolide) obtained from Cnidium officinale Makino] [Article in Japanese]Ozaki Y, Sekita S, Harada M.
The present study was carried out to elucidate a centrally acting musclerelaxant effect of chloroform soluble fraction and its component, namely, ligustilide, cnidilide and senkyunolide obtained from the rhizome of Cnidium officinale Makino. These three compounds were isolated from the chloroform soluble fraction by column chromatography on silica gel. The centrally acting muscle relaxant effect was investigated on the crossed extensor reflex in anesthetized rats and these samples were suspended in 0.5% carboxymethyl cellulose solution and administered i.p. These three compounds as well as the chloroform soluble fraction depressed the reflex response. The depressive potencies among them were almost the same and their potencies were also the same or somewhat weaker as that of mephenesin. As a curare-like action was not observed, a muscle relaxation induced by these phthalide compounds is considered to be due to central origin.PMID: 2810059 [PubMed - indexed for MEDLINE]

4: Clin Exp Pharmacol Physiol 1999 Oct;26(10):845-6

NBPA: a cerebral ischemic protective agent.

Zhang J, Peng X, Wei G, Su D.

1. NBPA is a derivative of 3-n-butylpathalide isolated from Apium granolens Linn.

2. At concentrations ranging from 6 x 10(-6) to 10(-6) mol/L, NBPA inhibited the L-type calcium current in guinea-pig myocardial cells and cultured human neuroblastoma cells.

3. At 10(-6) mol/L, NBPA markedly inhibited calcium-dependent and -independent release of glutamate from synaptosomes.

4. The [31P] nuclear magnetic resonance spectrum has shown that pretreatment with NBPA at 15 mg/kg, i.p., improved energy metabolism.

5. In situ hybridization has shown that 10 and 20 mg/kg, i.p., NBPA prior to cerebral artery occlusion can accelerate the expression of heat shock protein 70 mRNA and inhibit c-fos mRNA expression.

6. It has been shown that NBPA decreases the nitric oxide content and bc nitric oxide synthase (NOS) activity in the global cerebral ischaemia-reperfusion model in rats. In addition, it has been shown that NBPA significantly inhibits the expression of inducible NOS protein.

PMID: 10549420 [PubMed - indexed for MEDLINE]

5: Bioorg Med Chem 1999 Jul;7(7):1445-50

Structure-requirements of isocoumarins, phthalides, and stilbenes from

Hydrangeae Dulcis Folium for inhibitory activity on histamine release from rat

peritoneal mast cells.

Matsuda H, Shimoda H, Yoshikawa M.

We examined the structure-activity relationships of isocoumarins, phthalides and stilbenes isolated from Hydrangeae Dulcis Folium and related compounds for the inhibition of histamine release in rat peritoneal mast cells. The activities of isocoumarins such as thunberginols A and B were more potent than those of dihydroisocoumarins such as hydrangenol and thunberginol G. The double bond at the 3-position seemed to be essential to potentiate the activity. The hydroxyl groups at the 8-, 3'- and 4'-positions of isocoumarin were essential for the activity, while the hydroxyl group at the 6-position was scarcely needed. Since the activities of benzylidenephthalides such as thunberginol F were more potent than those of hydramacrophyllols A and B, the presence of a double bond at the 3-position was needed to increase the activity. Moreover, the hydroxyl group at the 8-position was essential for the activity. On the time course study, thunberginols A, B and F completely inhibited histamine release by pretreatment at 100 microM for 1 to 15 min, whereas DSCG inhibited histamine release only following 1-min pretreatment at 1000 microM. These results suggested that the mechanisms of the inhibitory effect of thunberginols are different from that of DSCG.

PMID: 10465418 [PubMed - indexed for MEDLINE]

6: Life Sci 1998;62(23):2073-82

Effects of methylenechloride-soluble fraction of Japanese angelica root extract, ligustilide and butylidenephthalide, on pentobarbital sleep in group-housed and socially isolated mice.

Matsumoto K, Kohno S, Ojima K, Tezuka Y, Kadota S, Watanabe H.

We previously showed that the extract of Japanese angelica root (JAR-E) reversed the decrease in pentobarbital (PB) sleep induced by isolation stress and yohimbine and methoxamine, stimulants of central noradrenergic systems, in mice. Here, we tested the effects of several fractions ffrom JAR-E and ligustilide an butylidenephthalide, phthalide components of JAR-E, on PB sleep in isolated mice to elucidate the mechanism of the action of JAR-E. Methanol-soluble (Met-S) and -insoluble (Met-IS) fractions were obtained from JAR-E. Methylenechloride-soluble (MC-S) and -insoluble fractions (MC-IS) were prepared from Met-S. MC-S (11.4-76 mg/kg, p.o.) reversed the isolation stress-induced decrease in PB sleep, but neither Met-IS (0.8-2.4 g/kg, p.o.) nor MC-IS (0.7-2 g/kg, p.o.) had the same effect. The i.p. administration of MC-S exhibited a similar activity to that observed after the p.o. administration of the same fraction. Ligustilide (5-20 mg/kg, i.p.) and butylidenephthalide (10-30 mg/kg, i.p.) reversed PB sleep decrease in isolated mice. Both components (20 mg/kg, i.p.) attenuated the suppressive effects of yohimbine (30 nmol, i.c.v.), methoxamine (200 nmol, i.c.v.) and a benzodiazepine inverse agonist FG7142 (10 mg/kg, i.p.) on PB sleep in group-housed mice. These results suggest the contribution of ligustilide and butylidenephthalide to the effect of JAR-E on PB sleep in isolated mice, and implicate central noradrenergic and/or GABA(A) systems in the effects of these components.

PMID: 9627086 [PubMed - indexed for MEDLINE]

7: Jpn J Pharmacol 1980 Feb;30(1):85-91

A newly isolated antispasmodic--butylidenephthalide.

Ko WC.

Butylidenephthalide (BdPh), ligustilide and butylphthalide were isolated and purified from neutral oil of Ligusticum wallichii Franch. Among these three, BdPh proved to be the most active in inhibiting rat uterine contractions induced by prostaglandin F2 alpha, oxytocin and ACh. In studies done to compare the effects of BdPh and papaverine (Pap), guinea pig ileum, vas deferens and taenia coli were used. BdPh inhibited contractile responses of the ileum to agonists including ACh, K+ and Ba2+ in normal Tyrode solution and to exogenous Ca2+ in high K+ (80 mM), Ca2+-free Tyrode solution, and also responses of vas deferens responses to norepinephrine. Thus, BdPh is a non-specific antispasmodic but weaker in potency than Pap. However, as the inhibitory effects of BdPh on phasic contraction (PC) and tonic contraction (TC) of preparations, including depolarized and non-depolarized ileum and taenia coli, were much the same, it is suggested that the action mechanism of BdPh may differ from that of Pap which inhibited TC more selectively than PC. It may be concluded that BdPh possesses an non-specific antispasmodic action like

Pap, the mechanism of action being different from that of Pap.

PMID: 7401411 [PubMed - indexed for MEDLINE]

8. Zhongguo Yao Li Xue Bao. 1999 Oct;20(10):929-33.

Effects of 3-n-butylphthalide on production of vasoactive substances by cerebral and aortic endothelial cells. Xu HL, Feng YP. Institute of Materia Medica, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100050, China. AIM:

The effects of dl-3-n-butylphthalide (dl-NBP), l-3-n-butylphthalide (l-NBP), and d-3-n-butylphthalide (d-NBP) on the production of nitric oxide (NO), epoprostenol (Epo) and endothelin-1 (ET-1) were investigated in cerebrovascular and aortic endothelium in culture. METHODS: Bovine cerebral endothelial cells (BCEC) and bovine aortic endothelial cells (BAEC) were cultured in Medium 199 in vitro. After incubation with dl-, l-, and d-NBP for 24 h, the release of NO, Epo, and ET-1 were analyzed by using spectrometry assay and radioimmunoassay (RIA) respectively. RESULTS: Low concentrations of dl- and l-NBP (0.1-10 mumol.L-1) enhanced nitrite and 6-ketoprostaglandin F1 alpha (6-ketoPGF1 alpha) production in both BAEC and BCEC after a 24-h incubation, and l-NBP has a potent effect on promoting Epo production in BCEC. The production of ET-1 secreted by BCEC and BAEC was increased after TNF alpha stimulation, this enhancement was not blunted by the simultaneous addition of dl-, l-, and d-NBP. CONCLUSION: 1) dl-NBP and l-NBP increase NO production in both BCEC and BAEC. 2) l-NBP increases more Epo production in BCEC than that in BAEC, and dl-NBP has selective effect on increasing Epo production in BCEC.

PMID: 11270994 [PubMed - indexed for MEDLINE]

9. Antioxidant, cyclooxygenase and topoisomerase inhibitory compounds from Apium graveolens Linn. seeds.

Momin RA, Nair MG. Department of Horticulture and National Food Safety and Toxicology Center, Michigan State University, East Lansing 48824, USA. Phytomedicine. 2002 May;9(4):312-8.

Cyclooxygenase inhibitory and antioxidant bioassay-directed extraction and purification of celery seeds yielded sedanolide (1), senkyunolide-N (2), senkyunolide-J (3), 3-hydroxymethyl-6-methoxy-2,3-dihydro-1H-indol-2-ol (4), L-tryptophan (6), and 7-[3-(3,4-dihydroxy-4-hydroxymethyl-tetrahydro-furan-2-yloxy)-4,5-dihydroxy-6-hydroxymethyl-tetrahydro-pyran-2-yloxy]-5-hydroxy-2-(4-hydroxy-3-methoxy-phenyl)-chromen-4-one (7). The structures of compounds 1-7 were determined using spectroscopic methods. Compound 4 is reported here for the first time. At 250 pg ml(-1), compounds 1-4, 6 and 7 displayed prostaglandin H endoperoxide synthase-I (COX-I) and prostaglandin H endoperoxide synthase-II (COX-II) inhibitory activities at pH 7. The acetylated product (5) of compound 4 also inhibited COX-I and COX-II enzymes when tested at 250 microg ml(-1). Compounds 6 and 7 exhibited good antioxidant activity at concentrations of 125 and 250 microg ml(-1). Only compounds 1-3 exhibited topoisomerase-I and -II enzyme inhibitory activity at concentrations of 100, 200 and 200 microg ml(-1), respectively.

PMID: 12120812 [PubMed - indexed for MEDLINE]

10. NSAID gastropathy: prevention by celery seed extracts in disease-stressed rats

Whitehouse M.W., Butters, DE, Clarke M L, Rainsford K D

Inflammopharmacology, Vol 9, No 1,2, pp 201 -209 (2001)

.