decline results from impaired neuronal plasticity
(Tapia A et al., 2008)
The pathogenesis of such neurodegenerative
condition still has not been established definitely
and involves multiple factors that influences several
systems, however it was known to be related to the
declining level of BDNF responsible for the loss of
the neuron function and structure.(Erickson et al.,
2012) BDNF has been known to have an important
roles in proliferation, differentiation, target
innervation, and survival of neurons of the central
and peripheral neuron system.(Tapia-Arancibia et
al., 2008). Lower levels of BDNF were also
associated with poorer memory. (Cunha, 2010)
Centella asiatica (CA), a small annual herb from
the family Apiaceae and native to Indonesia, India,
and many part of Asia has always been used widely
as a traditional medicines such as Aryuvedic
medicine, Chinese medicine, and many Southeast
Asian countries traditional medicine. (Lokanathan et
al., 2016). The CA, also known as pegagan in
Indonesia or gotu kola in India, have a significant
number of reviews on their medicinal uses along
with their supportive evidences (Gohil, Patel, &
Gajjar, 2010; Lokanathan et al., 2016; Orhan, 2012;
Rajakumari, 2010). Indicating the strong potential
of the plant in medicinal sector.
The primary active components of CA are
saponins (also called triterpenoid). (Singh &
Rastogi, 1969) Saponins include asiaticoside, a
trisaccharide with aglycone asiatic acid,
madecosside, and madasiatic acid. These
components are responsible for some of the CA
medicinal effects such as wound healing and
vascular effects. (Gohil et al., 2010)
CNS, cognitive, and antioxidant actions of CA
has been studied in many research and maybe due to
its brahmoside and brahminoside components, but
are yet to be confirmed by clinical studies. (Gohil et
al., 2010; Khotimah, Sumitro, Ali, & Widodo,
2015). Khotimah et al also found that methanolic
extract of CA could increase the BDNF level in
neuronal tissue of Rattus noevegicus strain Wistar
that were exposed to lipopolysaccharide.
(Khotimah, Riawan, & Kalsum, 2009)
However, there has been a gap in the current
research, in which study of the neuroprotective
effects of the CA has never been demonstrated on a
subject with aging. Using 20-24 months old Sprague
Dawley rats, we hypothesized that treatment with
CA extract could effectively induce a
neuroprotective effect on old age rats, hence this
study aims to expand the knowledge in CA
specifically its effectiveness to the level of BDNF
found on the brain tissue of aged rats.
2 METHODS
2.1 Study Design & Subjects
The Sprague-Dawley (SD) young (2-3-month-old)
and aged (20-24-month-old) male rats, were
obtained from Research and Development
Department of the Ministry of Health. In total, there
were 27 rats that were used in this study. SD is a
strain of albino rat which is used in many researches
because its calmness and ease of handling.
Before the experiment, the rats were
acclimatized for 1 week with lighting of 12 hours
(light on from 06:00 p.m. to 06:00 a.m.), constant
room temperature of 24
o
C, were given standard
food and ad libitum drink. The rats were divided
into 4 groups: the aged rats with no treatment as
negative control, the aged rats with treatment of CA
extract (300mg/kgBW), the aged rats with Vitamin
E treatment of 6 IU and the young rats. The Vitamin
E treatment were used as a positive control, whereas
the young rats were used to provide comparation
between aged and young rats. The animals were
kept for 28 days under the same environment where
the rats underwent acclimatization. The treatment
were given once every 7 days.
2.1.1 Daily Nutrients
The rats were fed with a type of pellets made from a
mixture of cornmeal, rice bran powder, fishmeal,
soybean, coconut, meat and bone meal, oat, ground
nut, canola, skimmed milk, and fish pellet with
brand names of SPA-Z and FF999. This standard
pellets contained 18.5%-20.5% protein, 4% fat, 6%
fiber, 8% ash, 0,9% calcium, 0,7% phosphor and
has metabolized energy of 3100-3200 kcal/kg. The
rats were given water, ad libitum.
2.2 Extraction of CA
The CA leaves were dried on a drying racks or
sundried until all the water content evaporated.
After being dried, the CA leaves were then grinded
until the leaves become powdery. Then, the grinded
CA leaves were extracted/macerated with an ethanol
solvent, until all the active constituents were
dissoluted into the solvent. This extraction process
were performed for 24-48 hours and then proceeded
with a separation of the active component from the
solvent. This was achieved by evaporation using
rotary evaporator. At last, the gravimetry analysis
were performed to analyze the water content so that
the solute percentage can be determined.
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