Supplementation of Turmeric Extract Does Not Improve

Neurological Function Following Repetitive Mild Traumatic Brain

Injury in the Rat

Andre Marolop Pangihutan Siahaan

and Iskandar Japardi

Department of Neurosurgery, Faculty of Medicine, Universitas Sumatera Utara, Jl. Dr. Mansyur No. 5, Medan, Indonesia

Keywords: Repetitive Traumatic Brain Injuries, Curcumin, Psychomotor Assessment.

Abstract: Turmeric has been in use since ancient times as a condiment and due to its medicinal properties. Curcumin,

the yellow coloring principle in turmeric, is a polyphenolic and a major active constituent. Besides the anti-

inflammatory, thrombolytic, and anti-carcinogenic, curcumin was also reported to have therapeutic potential

in Alzheimer’s disease by inhibiting the amyloid-β-protein aggregation. Inflammation and early degeneration

are two of main processes believed happened after repetitive mild traumatic brain injury. The effect of

curcumin was evaluated in weight drop model of repetitive mild traumatic brain injury. Male Sprague dawley

rats (n=10) were given multiple brain injury (40 gr mass drop from 1 m heights, 3 times daily on day 0,1,3,

and 7). Curcumin (500 mg/kg) were given orally. On the last day of injury, psychomotor assessment (beam

walk assessment and exit circle test) were performed. Control injured rats had a significant neurological deficit

(p<0.01). No significant different found control and treatment group. The study does not demonstrate the

efficacy of curcumin in rat with repetitive mild traumatic brain injury model.

1 INTRODUCTION

Traumatic brain injury (TBI) results from impact to

the head. The severity of this injury is varied, from

mild (brief and little change in consciousness or

mental status) to severe (prolonged loss of

consciousness and coma to fatal). Either mild of

severe TBI can result in short and long-term

disability. On a global scale, TBI is a serious health

concern and is the leading cause of mortality and

disability among individuals in young-age

population. TBI is one of the most common

neurological diagnoses in the US and the CDC has

estimated that 1.7 million people sustain TBI

annually (Rutland-Brown, 2006).

Repetitive mild TBI (rmTBI) is the form of TBI

that has gained public awareness, as well as within

military, scientific, and medical communities. TBI

accounts for around 28% of all combat casualties in

Iraq and Afghanistan (Okie, 2005). Persistent

accounts of rmTBI suffered by athletes have also

directed much attention. It is estimated that 1.8-3.8

million sports-related TBIs occur every year (Halstead

& Walter, 2010). About 60% of retired professional

football players sustained at least 1 concussion during

their careers and approximately 25% experienced

repeated injury (Guskiewicz, 2005).

Repetitive mild TBIs generally produce a

constellation of symptoms (e.g. headache, dizziness,

confusion) collectively known as post-concussive

syndrome (Halstead and Walter, 2010). Reports of

more serious consequences of rmTBI such as chronic

traumatic encephalopathy and increased co-morbidity

of neurodegenerative disorders (Omalu, 2010),

(Guskiewicz, 2007). This situation becomes even

more complex by the fact that rmTBI is extremely

difficult to detect. For the most part, routine imaging

approaches (CT and MRI) contribute little to the

evaluation and management of mild concussion

(Boven, 2009). The more advanced and specialized

approaches such as diffusion tensor imaging are

showing promise (Donald, 2011).

Histologically, there is protein aggregation

happened after rmTBI. The primary proteinopathy is

tau protein, in form of neurofribrillary tangle (NFT).

The most common secondary proteinopathies are

TDP-43 and amyloid-β-protein aggregation (McKee,

2013).

Turmeric (Curcuma longa) is a traditional

medicinal plant that also is commonly used as spice

in South as well as Southeast Asia. Curcumin, the

active ingredient of this plat, had been isolated since

long time ago and is considered as a potent

antiinflammation. In animal model, curcumin will be

bounded to beta structure in amyloid, so that reduces

486

Siahaan, A. and Japardi, I.

Supplementation of Turmeric Extract Does Not Improve Neurological Function Following Repetitive Mild Traumatic Brain Injury in the Rat.

DOI: 10.5220/0010075504860490

In Proceedings of the International Conference of Science, Technology, Engineering, Environmental and Ramiﬁcation Researches (ICOSTEERR 2018) - Research in Industry 4.0, pages

486-490

ISBN: 978-989-758-449-7

the plaque formation. Curcumin also will decrease the

aggregation process and stimulate the clearance.

Curcumin was also reported decreased the

accumulation of soluble NFT and inhibit kinases that

are involved in tau formation. Moreover, also in

animal model, curcumin supplementation was proved

to be effective in Alzheimer’s disease (Darvesh,

2012).

Until now, there is no proved effective therapy for

rmTBI. Curcumin, that was proved to be effective in

neurodegenerative disease, might potential to be

therapeutic agent in rmTBI. The aim of this study was

to prove the role of curcumin in neurological function

after rmTBI.

2 MATERIAL AND METHODS

2.1 Animal Model and Experimental

Groups

Male Sprague dawley rats (n=30) weighing 350 to

400 gr were housed in polycarbonate cages

maintained at 50 ± 10% humidity with a 12-hour light

and dark cycle. Rats were fed with standard

laboratory chow and water ad libitum. The

experimental protocol was approved by an

Institutional Review Committee of Universitas

Sumatera Utara, Medan, Indonesia. The animals were

also acclimatized to the laboratory condition prior to

experimentation for two weeks.

The rats were randomly allocated into three

groups (n=10) as following; a control (sham-

operated) group, a trauma group, and turmeric extract

(TE) groups. The rats in controlled group were placed

only in stereotactic apparatus, with neither trauma nor

drug treatment. The control group underwent trauma

protocol and the TE group underwent trauma protocol

and turmeric extract treatment

2.2 Drug Treatment

Turmeric extract (Sido Muncul, Semarang,

Indonesia) was given in 500 mg/kgBW dose,

suspended in 2 cc of double-distilled water. All rats

were weighed before the extract was given. The

extract was given orally via oral gavage every day for

consecutively eight days, at least two hours before

trauma protocol.

2.3 Weight Drop Brain Injury Model

Rats were placed under a stereotaxic frame. This

protocol was done without any anaesthesia. A 40-

gram weight was dropped from a height of 1 m unto

5 mm diameter pipe resting on the vertex. To prevent

skull fracture, a round metal with 3 cm diameter was

placed on the rats’ vertex. The trauma was given three

times daily, every 4 hours on day 0,1,3, and 7. Every

rat in control and TE groups underwent 12 traumas in

this research.

2.4 Outcome Assessment

2.4.1 Mortality and Body Weight

Death of the rats following trauma protocol were

noted. Every rat was weighed on day-0 before the

protocol started and on day -7, i.e on the last trauma

protocol.

2.4.2 Beam Walk Assessment

Rat was placed on a 1.5 m length plywood with 10 cm

width. For baseline data, we recorded the time needed

to cross the plywood and the maximal walking distant

before did the protocol. We also recorded maximal

time before rats fell down and marked whether there

was sign of disequilibrium. We repeated the

assessment

2.5 Statistical Analysis

The total time needed were reported in mean and

standard deviation. When comparisons were made

between groups, significance in between-group

variability was analysed using the one-way Anova

test with Tukey as post hoc test. Differences were

considered significant at the P <0.05.

3 RESULT

3.1 Mortality and Weight

There was no mortality in all groups. The rats still

survived, moved actively, and had good appetite after

three days protocol. We did not find significant

weight changes between day 0 and day 7 either in

negative sham control, trauma, or TE group (p>0.05;

table 1.

Supplementation of Turmeric Extract Does Not Improve Neurological Function Following Repetitive Mild Traumatic Brain Injury in the Rat

487

Table 1. Weight Change (gr)

Group Before traumas After traumas p

Negative sham

377.22 ± 29.72

378.44 ± 29.66

0,910

Trauma

351.78 ± 29.89 349.33 ± 38.90

0,482

367.89 ± 36.70 357.89 ± 39.89

0,950

One way Annova, significant if p<0.05

Table 2. Time needed to walk along the stick

Group n

x ± SD (s)

Negative sham 10

9,7 ± 1,95

0,342

Trauma 5

12,20 ± 3,89

TE 5

8,80 ± 1,30

One way Annova, significant if p<0.05

3.2 Disorder of Equilibrium

Repetitive TBI generated disorder of equilibrium. On

the negative sham control group, none of the rat

showed any sign of disequilibrium. On the trauma

group, 90% of the showed sign of disequilibrium one

day after trauma. On the TE group, 60% of the rats

showed the same sig, whether 30% among them

showed psychomotor depression, and only 10%

showed no sign of disequilibrium.

3.3 Walking Length and Fell Down

On negative sham control group, all of the animals

could walk for 1.5-metre long. Conversely, only 50%

of the animals in the trauma group that could walk for

1.5-metre long. 40% of them could not walk for 1.5

metre and 10% of them did not move at all. On the

TE group, 50% of animals could walk for 1.5-metre

length. 20% of the could not walk till the edge and

30% did not move at all.

The same finding could be seen regarding fell

down. None of the negative sham control group fell

down while walking on the plywood. Conversely,

40% animals on the control group fell down while

walking on the stick and 50% animals on the

treatment group fell down while walking.

Regarding the time needed to walk along the stick,

there was no significant different in all three groups

(table 2).

4 DISCUSSION

Repetitive mild traumatic brain injury is one of major

concerns in neurology right now. Many people are

prone to suffer this condition, such as athlete and

military personnel. In animal study, it is said that the

cellular as well as molecular balance will be back to

normal state in around 7-10 days (Giza and Hovda,

2014). This fact makes hypothesis arising, that brain

will be in prone-to-injury condition if the second

impact happened in this time period.

We used the modified weight drop model in this

study with modification from Marmarou procedure.

We showed zero mortality rate, compared to report by

Marmarou (64%) or by Kane (10%) (Marmarou,

1994), (Kane, 2012). Our finding was consistent with

the next study. They also reported the same mortality

rate (Xu, 2016). The other unique feature in our

model is no need of anesthesia. All the trauma

procedures were performed without anesthesia, even

this method could produce a non-uniform injury site.

Even so, this situation fitted contact injury happened

during sport.

We also did not find changes in body weight in all

groups following trauma. In many reports of animal

model of traumatic brain injury, there are significant

body weight decrease that probably happened due to

injury affecting feeding behavior and injury to the

anterior hypothalamus (Wei, 2012), (Samini, 2013),

(Moon, 2009).

We found significant different regarding motoric

and equilibrium. Rats in traumatic brain injury and

turmeric extract group could not maintain the stability

in the plywood. After trauma, only half of groups in

ICOSTEERR 2018 - International Conference of Science, Technology, Engineering, Environmental and Ramiﬁcation Researches

488

either trauma or TE group that could maintain the

stability. A study reported that motoric disfunction

was one of common manifestation in Chronic

traumatic encephalopathy, a sequel of repetitive mild

traumatic brain injury (Montenigro, 2015).

Turmeric extract is a well-known potent

antioxidant and antiinflammation. On animal study,

many reports showed the effectivity in degenerative

disease, but it was failed in human clinical study

(Darvesh, 2012), (Tang, 2017). In our study, we did

not find significant different regarding outcome in

trauma and turmeric extract group. One of the main

problems of crude turmeric extract is the low

bioavailability. If the challenge of the low

bioavailability is overcome, curcumin as medication

in repetitive mild traumatic brain injury may still be

in horizon.

The main limitation of this study is the outcome

that only limited to clinical. For the next research, it

is advised to do biochemistry analysis to determine

the cellular status and cell expression. Besides, a long

follow up period is advisable to make sure the long-

term outcome in this condition.

In conclusion, we found no significant

improvement after curcumin supplementation in

repetitive mild traumatic brain injuries.

ACKNOWLEDGEMENTS

This research was fully funded by Research

Committee of Universitas Sumatera Utara in

TALENTA Grant 2017.

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