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Clitoria ternatea root extract enhances acetylcholine content in rat

hippocampus

Article in Fitoterapia · January 2003

DOI: 10.1016/S0367-326X(02)00249-6 · Source: PubMed

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 Fitoterapia 73 (2002) 685–689

Clitoria ternatea root extract enhances

acetylcholine content in rat hippocampus
K.S. Raia,*, K.D. Murthyb, K.S. Karanthc, K. Nalinid,
M.S. Raoe, K.K. Srinivasanf
a
Department of Physiology, Kasturba Medical College, Manipal Pin 576119, Karnataka, India
b
Department of Physiology, International Centre for Health Sciences, Manipal Pin 576119, Karnataka,
India
c
Department of Pharmacology, K.S. Hegde Medical Academy, Deralkatte, Mangalore, Karnataka,
India
d
Department of Clinical Biochemistry, Kasturba Medical College, Manipal Pin 576119, Karnataka,
India
e
Department of Anatomy, Kasturba Medical College, Manipal Pin 576119, Karnataka, India
f
Department of Chemistry, College of Pharmaceutical Sciences, Manipal Pin 576119, Karnataka,
India

Received 5 October 2001; accepted in revised form 26 September 2002

Abstract

Treatment with 100 mgykg of Clitoria ternatea aqueous root extract (CTR), for 30 days
in neonatal and young adult age groups of rat, significantly increased acetylcholine (ACh)
content in their hippocampi as compared to age matched controls. Increase in ACh content
in their hippocampus may be the neurochemical basis for their improved learning and
memory.
䊚 2002 Elsevier Science B.V. All rights reserved.

Keywords: Clitoria ternatea; Acetylcholine (ACh); Hippocampus

1. Introduction

In the Ayurvedic system of medicine, many herbs and plants are used to treat
various ailments. ‘Medhya’ drugs mentioned in Ayurvedic texts are a group of
*Corresponding author. Tel.: q91-8252-71201, ext. 22321; fax: q91-8252-70500y70062.
E-mail addresses: kiranmai_rai2001@yahoo.com (K.S. Rai),
kiranmai_rai2002@hotmail.com (K.S. Rai).

0367-326X/02/$ - see front matter 䊚 2002 Elsevier Science B.V. All rights reserved.
PII: S 0 3 6 7 - 3 2 6 X Ž 0 2 . 0 0 2 4 9 - 6
686 K.S. Rai et al. / Fitoterapia 73 (2002) 685–689

herbal medicines, used to improve mental abilities w2x. These herbal drugs include
extracts from Clitoria ternatea w3,4x, Celastrus panniculatus, Acorus calamus,
Centella asiatica, Withania somnifera, Guduchi and Areca w4–6x.
C. ternatea, commonly called Shankapushpi, is a tropical twining herb, growing
wild and also in gardens, bearing whiteyink blue flowers resembling a conch-shell
w3,4x.
Extracts of this plant have been used as an ingredient in ‘Medhya Rasayana’ a
rejuvenating recipe used for treatment of neurological disorders and considered as
wholesome for intellect w7x. Oral intubation with 100 mgykg of aqueous root extract
of C. ternatea for 30 days has proved to improve learning and memory in rats.
Further work on the dendritic arborization of CA3 pyramidal neurons in the
hippocampi of rats showed significant increase in apical and basal dendritic branches
w1,8x. Intraperitoneal administration of alcoholic extract of C. ternatea to rats and
mice has been reported to produce sedation and diminished alertness w9x. Oral
treatment with alcoholic extracts of aerial and root parts of C. ternatea has been
reported to increase ACh content and acetylcholinesterase activity in the rat brain
and improve memory retention w10x. The present study was aimed to estimate the
ACh content in the hippocampi of rats treated with aqueous root extract of C.
ternatea.

2. Experimental

2.1. Plant material

C. ternatea roots collected from 2–3-year-old plants (bearing white flowers),

identified and confirmed as C. ternatea in the Department of Pharmacognosy,
College of Pharmaceutical Sciences, Manipal, were used for extraction.

2.2. Extract preparation

Aqueous root extract of C. ternatea was prepared, as reported earlier w1x and the
dry extract stored in an airtight container was used for the study. Preliminary
phytochemical screening w11,12x of the aqueous extract of C. ternatea gave positive
tests for amino acids, proteins and carbohydrates and negative tests for alkaloids,
sapoins, flavonoids, coumarins and lignans.

2.3. Animals

Wistar rats of either sex of two age groups, (i) neonatal pups (7 days old) and
(ii) young adults (60 days old) were used for the study. These rats were housed
under standard laboratory conditions with 12-h lightydark cycle. The animals were
fed with pellet chow and water ad libitum.

2.4. Drug treatment

Seven-day-old neonatal rat pups and 60-day-old young adult rats were further
subdivided into two groups. Control (C), orally treated with 10 mlykg of saline and
 K.S. Rai et al. / Fitoterapia 73 (2002) 685–689 687

orally treated (CTR 100) with 100 mgykg of aqueous root extract of C. ternatea
for 30 days (from 8th day to 37th day in neonatal group and from 61st day to 90th
day in young adult rats).

2.5. Tissue preparation

After the period of treatment, both neonatal and young adult rats (C and CTR
100 groups) were killed by quick decapitation. Their brains were rapidly removed
and placed on ice. Hippocampi were then dissected out on ice and placed in chilled
0.9% NaCl solutions. Hippocampi from three rats were pooled for estimation of
ACh (ns9 pooled samples in each group).

2.6. ACh estimation

ACh was estimated by fluorimetric method w13x. The pooled hippocampi were
weighed and homogenized using a Teflon–glass homogenizer in freshly prepared
cold 10% trichloroacetic acid.
The homogenates were centrifuged at 10 000 rev.ymin for 10 min at 4 8C. The
supernatant was collected and processed immediately for estimation of ACh.

2.7. Statistical analysis

Results, expressed as mean"S.D., were statistically compared by Student’s t-test.

Values of P-0.05 were considered statistically significant.

3. Results and discussion

Oral treatment with 100 mgykg of aqueous root extract of C. ternatea, during
neonatal period and young adult age in rats, enhanced ACh content significantly in
their hippocampi: neonatal rats from 52.79"12.36 to 68.83"9.87 nmolyg tissue,
young adults rats from 33.9"6.92 to 88.89"18.29 nmolyg tissue (Fig. 1).
Hippocampal ACh content was found to be significantly less in 90-day-old control
rats as compared to 37-day-old control rats: (33.9"6.92 vs. 52.79"12.36 nmolyg
tissue). On the contrary, hippocampal ACh content was found to be higher in 90-
day-old CTR treated rats than in 37-day-old CTR treated rats 88.89"18.29 tissue
vs. 68.83"9.87 nmolyg tissue.
In our earlier report w8x we have shown that C. ternatea aqueous root extract
treatment to rats induced an increase in the dendritic arborization of CA3 hippocam-
pal neurons, suggesting an increase in protein synthesis in this region. Thus, we can
speculate that C. ternatea aqueous root extract treatment may have increased the
ACh synthetic enzymes.
The ability of C. ternatea aqueous root extract treatment in rats to increase ACh
content suggests that this extract treatment may be useful as a memory enhancer,
688 K.S. Rai et al. / Fitoterapia 73 (2002) 685–689

Fig. 1. Effect of C. ternatea on acetylcholine content in rat’s hippocampi. Each bar represents
mean"S.D. wStudent’s t-test significance (control vs. CTR treated) shown in parentheses.x Ns9.

similar to synthetic memory enhancing drugs like Piracetam w14x, Nefiracetam w15x,
dehydroepiandrosterone sulfate w16x and FG7142 w17x.
It may be concluded that C. ternatea aqueous root extract treatment may be of
value for reinforcing depressed cholinergic transmission in certain age related
memory disorders and to improve learning and memory in normal individuals.
Mechanism of its action needs further exploration.

Acknowledgments

The authors gratefully acknowledge the financial support provided by Kasturba

Medical College, a constituent college of Manipal Academy of Higher Education
(MAHE-Deemed University), Manipal, Karnataka, India. The authors also thank
Mr. Harish Bhasker of Kirah Graphics, Manipal, for his help in the preparation of
this manuscript.

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