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Evaluation of anxiolytic activity of spray dried powders of two South Brazilian Passiflora species.

The Passiflora extracts have been used in folk medicine because of its reputed sedative and anxiolytic properties. The present study aimed to compare the potential anxiolytic activity of two Passiflora spray-dried powders obtained from P. alata and P. edulis, known in Brazil as 'maracujá'. Male adult Swiss rats were treated with 200, 400 and 800 mg/kg of spray-dried powders p.o. and anxiolytic activity was evaluated using the elevated plus-maze test. The spray-dried powders showed anxiolytic activity in doses of 400 and 800 mg/kg. Our results support the potential anxiolytic effect of Passiflora spray-dried powders (P. alata and P. edulis).

Administration, Oral↗

Neuropharmacological evaluation of the putative anxiolytic effects of Passiflora edulis Sims, its sub-fractions and flavonoid constituents.

Passiflora edulis Sims together with several other plants of the genus Passiflora have been reported to possess anxiolytic properties. It has been suggested recently that flavonoids may be partly responsible for the neuropharmacological activity of these plants but there are still few data reporting the relation between the constituents of these plants and their activity. This work evaluated the anxiolytic/sedative activity of an aqueous extract of Passiflora edulis Sims and bioguided its fractionation using the elevated plus-maze model of anxiety and other complementary pharmacological tests. The aqueous extract presented an anxiolytic-like activity without any significant effect upon the motor activity whilst the total flavonoid fraction (TFF) presented an anxiolytic-like activity but compromised motor activity. Through fractionation of TFF it was possible to isolate and characterize luteolin-7-O-[2-rhamnosylglucoside] which showed an anxiolytic-like activity without compromising motor activity.

Animals↗

Passiflora actinia Hooker extracts and fractions induce catalepsy in mice.

Leaves from several Passiflora species are largely employed in the Brazilian folk medicine for its anxiolytic and sedative properties. In behavioral studies, to analyze the tranquillizer action of Passiflora actinia Hooker, it was noteworthy that animals treated with the hydroethanol (HE) and methanol (ME) extracts presented an abnormal postural immobility compared to control animals. That observation led to an investigation of the effects of HE and its fractions on evaluation of catalepsy in mice. The results showed that HE extract, methanol extract, the sequent aqueous crude fractions (AF), and fa, fb and fc chromatographic fractions obtained from Passiflora actinia induced catalepsy in mice. Apparently, the active principles responsible for catalepsy are present in all of the fractions of the extract.

Animals↗

Phylogenetic relationships among Passiflora species based on the glutamine synthetase nuclear gene expressed in chloroplast (ncpGS).

This paper presents the first molecular phylogeny of the genus Passiflora encompassing almost all sections of this large genus. The nuclear-encoded chloroplast-expressed glutamine synthetase gene (ncpGS) was used to examine the relationships among Passiflora species (passionflowers), which was then compared with the new classification proposed by Feuillet and MacDougal. The resulting Bayesian, likelihood, and parsimony trees are congruent and well supported. The 90 Passiflora species examined apparently split into eight main subgenera: Plectostemma, Granadilla, Astrophea, Deidamioides, Polyanthea, Dysosmia, Tetrapathea, and Tryphostemmatoides. These results are in overall agreement with the Feuillet and MacDougal's classification but here we propose that three additional subgenera, Polyanthea, Dysosmia, and Tetrapathea, should be maintained. We observe a striking overall correlation between the phylogenetic position of the different species and their chromosome number. The first clade contains the arborescent species of the subgenus Astrophea, with n=12. The second clade, subgenus Plectostemma, includes species from four subgenera of Killip's classification with n=6 chromosomes. The last clade, subgenus Granadilla, includes species of seven old subgenera with n=9. Subgenus Dysosmia, with a variable chromosome number of n=9-11, is considered here as a separate subgenus closely related to the subgenus Granadilla.

Cell Nucleus↗

[Passion Flower (Passiflora incarnata L.)--a reliable herbal sedative].

Extracts and fluid extracts from the aerial parts from Passiflora incarnata L. are widely used as components of herbal sedatives. Many pharmacological investigations confirm the sedative effects of Passiflorae herba. From some of the studies also anxiolytic effects can be deduced. As Passionflower is mainly used in combinations, clinical studies of the single drug are not available. Based on pharmacological data, the experiences of traditional use and the use in combinations Passiflora extracts are an important factor in the phytotherapy of tenseness, restlessness and irritability with difficulty in falling asleep.

Anxiety↗

Hypnotic activities of chamomile and passiflora extracts in sleep-disturbed rats.

In the present study, we investigated hypnotic activities of chamomile and passiflora extracts using sleep-disturbed model rats. A significant decrease in sleep latency was observed with chamomile extract at a dose of 300 mg/kg, while passiflora extract showed no effects on sleep latency even at a dose of 3000 mg/kg. No significant effects were observed with both herbal extracts on total times of wakefulness, non-rapid eye movement (non-REM) sleep and REM sleep. Flumazenil, a benzodiazepine receptor antagonist, at a dose of 3 mg/kg showed a significant antagonistic effect on the shortening in sleep latency induced by chamomile extract. No significant effects were observed with chamomile and passiflora extracts on delta activity during non-REM sleep. In conclusion, chamomile extract is a herb having benzodiazepine-like hypnotic activity.

Animals↗

Isolation, culture, and plant regeneration from leaf protoplasts of Passiflora.

The family Passifloraceae contains many species exploited in the food, pharmaceutical, and ornamental plant industries. The routine culture of isolated protoplasts (naked cells) followed by reproducible plant regeneration, is crucial to the genetic improvement of Passiflora spp. by somatic cell technologies. Such procedures include somatic hybridization by protoplast fusion to generate novel hybrid plants, and gene introduction by transformation. Seedling leaves are a convenient source of totipotent protoplasts. The protoplast-to-plant system developed for Passiflora edulis fv. flavicarpa is summarized in this chapter. The procedure involves enzymatic degradation of leaf tissue using commercially-available Macerozyme R10, Cellulase R10, and Driselase. Isolated protoplasts are cultured in Kao and Michayluk medium, semi-solidified with agarose. The medium containing the suspended protoplasts is dispensed as droplets or thin layers and bathed in liquid medium of the same composition. Shoot regeneration involves transfer of protoplast-derived tissues to Murashige and Skoog-based medium. The protocols developed for P. edulis are applicable to other Passiflora spp. and will underpin the future biotechnological exploitation of a range of species in this important plant family.

Cell Culture Techniques↗

Cholesterol-dependent hemolytic activity of Passiflora quadrangularis leaves.

Plants used in traditional medicine are rich sources of hemolysins and cytolysins, which are potential bactericidal and anticancer drugs. The present study demonstrates for the first time the presence of a hemolysin in the leaves of Passiflora quadrangularis L. This hemolysin is heat stable, resistant to trypsin treatment, has the capacity to froth, and acts very rapidly. The hemolysin activity is dose-dependent, with a slope greater than 1 in a double-logarithmic plot. Polyethylene glycols of high molecular weight were able to reduce the rate of hemolysis, while liposomes containing cholesterol completely inhibited it. In contrast, liposomes containing phosphatidylcholine were ineffective. The Passiflora hemolysin markedly increased the conductance of planar lipid bilayers containing cholesterol but was ineffective in cholesterol-free bilayers. Successive extraction of the crude hemolysin with n-hexane, chloroform, ethyl acetate, and n-butanol resulted in a 10-fold purification, with the hemolytic activity being recovered in the n-butanol fraction. The data suggest that membrane cholesterol is the primary target for this hemolysin and that several hemolysin molecules form a large transmembrane water pore. The properties of the Passiflora hemolysin, such as its frothing ability, positive color reaction with vanillin, selective extraction with n-butanol, HPLC profile, cholesterol-dependent membrane susceptibility, formation of a stable complex with cholesterol, and rapid erythrocyte lysis kinetics indicate that it is probably a saponin.

Animals↗

Toxicity of Passiflora incarnata L.

BACKGROUND: Herbal medicines may have significant adverse effects which are not suspected or recognized. CASE REPORT: A 34-year-old female developed severe nausea, vomiting, drowsiness, prolonged QTc, and episodes of nonsustained ventricular tachycardia following self-administration of a herbal remedy, Passiflora incarnata L., at therapeutic doses. The possible association of symptoms with passiflora was not recognized for several days. She required hospital admission for cardiac monitoring and intravenous fluid therapy. CONCLUSIONS: Passiflora incarnata was associated with significant adverse effects in this patient. It is important to ask specifically about the use of herbal medicines in patients with undiagnosed illnesses.

Adult↗

Correct Identification of Passiflora incarnata Linn., a Promising Herbal Anxiolytic and Sedative.

Passiflora incarnata Linn. and Passiflora edulis Sims are the two important plants of the family Passifloraceae that have often been reported as synonymous because of their identical morphological and microscopic characteristics. P. incarnata is a popular sedative and anxiolytic, whereas, P. edulis is rarely reported to possess significant central nervous system depressant activity. P. edulis, as the name of the species reflects, is mainly grown for edible purposes. During a survey of literature on the genus Passiflora, it was noticed that in many references the two plants are mentioned synonymously. The designation by Sir William J. Hooker in 1843, followed by the citation of P. edulis as the synonym of P. incarnata in Index Kewensis of 1895, not only substantiated the controversial identity but also caused confusion to researchers. The prevailing confusion might have led to improper selection of the bioactive plant, thereby accounting for inconclusive and contradictory pharmacological reports on either of the two plants. In this work, we establish key identification parameters to differentiate P. incarnata from P. edulis. Various leaf constants such as vein-islet number, vein-termination number, stomatal number, and stomatal index are different for the two species. Physicochemical parameters such as ash values and extractive values and the thin layer chromatography profile of the petroleum ether extract of P. incarnata and P. edulis are also distinct and different. Various clinical uses of P. incarnata for anxiety and allied diseases are discussed.

Journal Article↗

Molecular investigations of pathogenesis-related Bet v 1 homologues in Passiflora (Passifloraceae).

The major birch pollen allergen, Bet v 1, responsible for allergic reactions in many areas of the world, is homologous to a large number of pathogenesis-related proteins (PRs), identified as PR10. As part of a long-range investigation of these types of proteins and of evolution in Passiflora, DNA sequences from eight Bet v 1 homologue isoforms were obtained from five species of this genus in Brazil, and their sequences compared among themselves and with 30 others from 8 different species, classified in different taxonomic units. The objective was a first characterization of these PRs in wild passionflowers, and their use for evolutionary and applied investigations. High interspecific, but low intraspecific variability was observed, as expected from multigenic families subjected to concerted evolution. The relationships obtained both within Passiflora and between it and seven other genera probably best reflect functional similarities than evolutionary history.

Allergens↗

Differential organellar inheritance in Passiflora's (Passifloraceae) subgenera.

Four chloroplast (cp), one mitochondrial (mt), and one ribosomal nuclear (ITS) DNA regions were studied in four artificial and one natural interspecific Passiflora hybrids. The ITS results confirmed their hybrid origin and all mtDNAs were maternally inherited. The same, however, was not true for cpDNA. The four hybrids (three artificial and one natural) derived from species of the Passiflora subgenus showed a cpDNA paternal inheritance, while the one involving taxa of the Decaloba subgenus gave evidence of maternal transmission. These results are of significance for the ongoing studies which are being performed on the molecular evolution of this genus and furnish important background for investigations aimed at clarifying the factors which determine cpDNA inheritance.

Base Sequence↗

Possible involvement of GABA A-benzodiazepine receptor in the anxiolytic-like effect induced by Passiflora actinia extracts in mice.

Hydroethanol (HE) and methanol (ME) extracts obtained from the leaves of Passiflora actinia Hooker were evaluated for behavioral effects in mice. Single-dose oral administration of HE (300 and 600 mg/kg) or ME (100 and 300 mg/kg) resulted in anxiolytic-like effects in the elevated plus-maze. The anxiolytic-like effects were also seen after the repeated administration of the HE (100 and 300 mg/kg). Flumazenil (10mg/kg, i.p.), a GABA(A)-benzodiazepine receptor antagonist, blocked the effects of ME (300 mg/kg, p.o.) and HE (600 mg/kg). At higher doses, a sedative effect produced by acute administration of HE (600 mg/kg) or ME (300 mg/kg) was indicated by the potentiation of pentobarbital-induced sleep. With regard to memory-disrupting effects of anxiolytics, mice were evaluated by measuring the retest step-down latency 24h after foot-shock in a passive avoidance task. In contrast to diazepam (0.5mg/kg) or piracetam (200mg/kg), ME (30, 100 and 300 mg/kg) or HE (100, 300 and 600 mg/kg) did not influence the step-through latency in the acquisition or retention memory tasks. The present results show an anxiolytic profile for HE and ME of Passiflora actinia. There are also indications of an involvement of GABA(A) system in this effect.

Administration, Oral↗

LC and UV determination of flavonoids from Passiflora alata medicinal extracts and leaves.

The leaves of Passiflora alata Curtis (an official species in the Brazilian Pharmacopoeia) are used to treat anxiety, as antispasmodics and sedatives. The development of new analytical methods for analysing flavonoids in Passiflora species is needed in order to improve the quality assurance of the plant and derived extracts and phytomedicines. A survey on the profile of flavonoids and content of isovitexin in the leaves and fluidextract of P. alata through LC was carried out, comparing its chromatographic profiles with a commercial P. incarnata fluidextract. Also, the concentration of the total flavonoids of extracts and leaves according to phamarcopoeial photometrical methods was determined and discussed. The fluidextract of P. alata was produced in accordance with the Pharmacopoeia Helvetica method; the presence of isovitexin in both species (which have distinct chromatographic profiles) was evidenced, this being the major flavonoid compound in the P. incarnata (1.198 g%), but not in P. alata (0.018 g%) fluidextract. Only traces of vitexin could be observed in the P. alata, as well as the absence of the other tested flavonoids: orientin and swertisin (P. incarnata markers), hyperoside, rutin, hesperidin and clorogenic acid. The LC developed method was shown to be appropriated for the qualitative and quantitative analyses and for differentiate species, suitable to be applied to commercial sample analysis. The spectrophotometrical results of three different methods described for P. incarnata were not comparable, the best performance being the British Pharmacopoeia method.

Chromatography, Liquid↗

Cyanohydrin glycosides of Passiflora: distribution pattern, a saturated cyclopentane derivative from P. guatemalensis, and formation of pseudocyanogenic alpha-hydroxyamides as isolation artefacts.

Nineteen species of Passiflora (Passifloraceae) were examined for the presence of cyanogenic glycosides. Passibiflorin, a bisglycoside containing the 6-deoxy-beta-D-gulopyranosyl residue, was isolated from P. apetala, P. biflora, P. cuneata, P. indecora, P. murucuja and P. perfoliata. In some cases this glycoside co-occurs with simple beta-D-glucopyranosides: tetraphyllin A, deidaclin, tetraphyllin B, volkenin, epivolkenin and taraktophyllin. P. citrina contains passicapsin, a rare glycoside with the 2,6-dideoxy-beta-D-xylo-hexopyranosyl moiety, while P. herbertiana contains tetraphyllin A, deidaclin, epivolkenin and taraktophyllin, P. discophora tetraphyllin B and volkenin, and P. x violacea tetraphyllin B sulfate. The remaining species were noncyanogenic. The glycosides were identified by 1H and 13C NMR spectroscopy following isolation by reversed-phase preparative HPLC. From P. guatemalensis, a new glucoside named passiguatemalin was isolated and identified as a 1-(beta-D-glucopyranosyloxy)-2,3-dihydroxycyclopentane-1-carbonitrile. An isomeric glycoside was prepared by catalytic hydrogenation of gynocardin. alpha-Hydroxyamides corresponding to the cyanogenic glycosides were isolated from several Passiflora species. These alpha-hydroxyamides, presumably formed during processing of the plant material, behave as cyanogenic compounds when treated with commercial Helix pomatia crude enzyme preparation. Thus, the enzyme preparation appears to contain an amide dehydratase, which converts alpha-hydroxyamides to cyanohydrins that liberate cyanide; this finding is of interest in connection with analysis of plant tissues and extracts using Helix pomatia enzymes.

Amides↗

Variability of the 5S and 45S rDNA sites in Passiflora L. species with distinct base chromosome numbers.

Cytologically, the species of Passiflora with known chromosome number can be divided into four groups: (1) 2n = 12, 24, 36; (2) 2n = 24; (3) 2n = 18, 72; and (4) 2n = 20. The base chromosome number proposed for the genus is x = 6, with x = 9, x = 10 and x = 12 being considered secondary base numbers. In the present study, variability of 5S and 45S rDNA sites was investigated in 20 species of these four groups to check the reliability of this hypothesis. In the group with x = 6, five diploid species (2n = 12) exhibit two 5S rDNA sites and two (P. capsularis, P. morifolia and P. rubra) or four (P. misera 2x and P. tricuspis) 45S rDNA sites. The hexaploid cytotype of P. misera had 12 45S rDNA sites and six 5S rDNA. A tetraploid species, P. suberosa, had ten 45S rDNA sites and four 5S rDNA sites, both in the same chromosomes as the 45S rDNA sites. In the group with x = 9, P. actinia, P. amethystina, P. edmundoi, P. elegans, P. galbana, P. glandulosa and P. mucronata displayed six 45S rDNA sites, whereas P. alata, P. cincinnata, P. edulis f. flavicarpa, P. edulis var. roxo and P. laurifolia had four sites. In this group, all species were diploid (2n = 18) and had only two 5S rDNA sites. Passiflora foetida, the only species with 2n = 20, had six 45S rDNA sites and four 5S rDNA sites. The species with x = 12 (2n = 24), P. haematostigma and P. pentagona, showed four 45S rDNA sites and two 5S rDNA. In general, the number and location of 5S and 45S rDNA sites were consistent with the hypothesis of x = 6 as the probable ancestral genome for the genus, while the groups of species with x = 9, x = 10 and x = 12 were considered to be of tetraploid origin with descending dysploidy and gene silencing of some redundant gene sites, mainly those of 5S rDNA.

Chromosomes, Plant↗

Isolation and characterization of a myo-inositol-1-phosphate synthase gene from yellow passion fruit (Passiflora edulis f. flavicarpa) expressed during seed development and environmental stress.

BACKGROUND AND AIMS: Myo-inositol-1l-phosphate synthase (MIPS) catalyses the conversion of d-glucose 6-phosphate to 1-l-myo-inositol-1-phosphate, the first and rate-limiting step in the biosynthesis of all inositol-containing compounds. Inositol phospholipids play a vital role in membrane trafficking and signalling pathways, auxin storage and transport, phytic acid biosynthesis, cell wall biosynthesis and production of stress-related molecules. In the present study, an MIPS cDNA from developing Passiflora edulis f. flavicarpa seeds was characterized and an investigation made into its spatial and differential expression, as well as changes in its transcription during exposure of growing plants to cold and heat stresses. METHODS: The MIPS-encoding gene was isolated by polymerase chain reaction (PCR) methods, and transcript levels were examined using semi-quantitative reverse transcription-PCR (RT-PCR) during seed development and in response to heat and cold stress. In addition, the copy number of the cloned PeMIPS1 gene in the genome of Passiflora edulis, P. eichleriana, P. caerulea, P. nitida and P. coccinea was determined by Southern blot analyses. KEY RESULTS: A full-length cDNA clone of the PeMIPS1 from P. edulis was isolated and characterized. Southern blot analyses indicated that the genomic DNA might have diverse sequences of MIPS-encoding genes and one copy of the cloned PeMIPS1 gene in the genomes of P. edulis, P. eichleriana, P. caerulea, P. nitida and P. coccinea. RT-PCR expression analyses revealed the presence of PeMIPS1 transcripts in ovules, pollen grains and leaves, and during the seed developmental stages, where it peaked at 9 d after pollination. The PeMIPS1 gene is differentially regulated under cold and heat stress, presenting a light-responsive transcription. CONCLUSIONS: Experimental data suggest that PeMIPS1 transcription plays an important role in the establishment of developmental programmes and during the response of plants to environmental changes. The PeMIPS1 is differentially transcribed during cold and heat stress, presenting a light response pattern, suggesting that it is important for environmental stress response.

Cloning, Molecular↗

RAPD-based genetic linkage maps of yellow passion fruit (Passiflora edulis Sims. f. flavicarpa Deg.).

A single cross between two clones of passion fruit (Passiflora edulis Sims. f. flavicarpa Deg., 2n = 18) was selected for genetic mapping. The mapping population was composed of 90 F1 plants derived from a cross between 'IAPAR 123' (female parent) and 'IAPAR 06' (male parent). A total of 380 RAPD primers were analyzed according to two-way pseudo-testcross mapping design. The linkage analysis was performed using Mapmaker version 3.0 with LOD 4.0 and a maximum recombination fraction (theta) of 0.30. Map distances were estimated using the Kosambi mapping function. Linkage maps were constructed with 269 loci (2.38 markers/primer), of which 255 segregated 1:1, corresponding to a heterozygous state in one parent and null in the other. The linkage map for 'IAPAR123' consisted of 135 markers. A total of nine linkage groups were assembled covering 727.7 cM, with an average distance of 11.20 cM between framework loci. The sizes of the linkage groups ranged from 56 to 144.6 cM. The linkage map for 'IAPAR 06' consisted of 96 markers, covering 783.5 cM. The average distance between framework loci was 12.2 cM. The length of the nine linkage groups ranged from 20.6 to 144.2 cM. On average, both maps provided 61% genome coverage. Twenty-four loci (8.9%) remained unlinked. Among their many applications, these maps are a starting point for the identification of quantitative trait loci for resistance to the main bacterial disease affecting passion fruit orchards in Brazil, caused by Xanthomonas campestris pv. passiflorae, because parental genotypes exhibit diverse responses to bacterial inoculation.

Chromosome Mapping↗