Effect of 10% Lavender Essential Oil Balm on Serum Cortisol Levels

in Male Wistar Rats

Christian Jaya Sumarto Putra

Institute Faculty of Medicine, Widya Mandala Catholic University Surabaya, Surabaya, Indonesia

Keywords: Lavender, Essential Oil, Balm, Serum Cortisol, Wistar Rat.

Abstract: Lavender (Lavandula angustifolia) is a Mediterranean plant that has developed throughout the world and

often used as aromatherapy for relaxation. Balm is one of the topical drug forms that effective for patients

with very dry skin and has a higher potency and greater drug penetration. Therefore, this study was

conducted to determine the effect of 10% lavender essential oil balm on serum cortisol levels. This study

used 36 male Wistar rats divided into 4 groups (negative control = no stressor; positive control = stressor

only; placebo = stressor + placebo; L

= stressor + Lavender 10%), the forced swim test was given as the

stressor. Serum cortisol levels were analysed using the Kruskal-Wallis Test (p<0.05) and continued with

the Mann-Whitney Test (p<0,05). The result of the serum cortisol levels analysis showed that 10%

lavender essential oil balm significantly (p=0.007 and p=0.041) decreased the serum cortisol levels in rats

compared to negative control group and positive control group (684.19 ± 54.081 (L

), 712.95 ± 129.589 (C

), and 728.13 ± 48.125 (C

)). These results indicate that lavender essential oil balm can be used as an

alternative treatment to relieve stress but should be further researched for other biochemical parameters.

1 INTRODUCTION

The incidence of stress is still high in various groups

and professions in the world. According to the World

Health Organization, more than 300 million people

in 2015 suffer from depression and depression is the

leading cause of morbidity and disability in the

world (World Health Organization, 2017). In 2013,

the Indonesian Ministry of Health published Basic

Health Research (Riskesdas) stated that 6% of the

total population in Indonesia experienced emotional

mental disorder (Kementerian Kesehatan, 2013).

Riskesdas Data in 2018 showed that the figure had

increased to 9.8% (Kementerian Kesehatan, 2018).

Untreated stress can lead to various problems.

According to a study conducted by Wada et al.

(2013) stated that high occupational stress exposure

could lead to the onset of depression (Wada et al.,

2013). Furthermore, patients diagnosed with acute

stress reactions had a greater rate for

completed suicide (Gradus et al., 2010). Stress is

a stimulus that evokes the release of ACTH and

adrenal glucocorticoid (Fink, 2016). Stress causes

activation of various physiological responses

especially in the endocrine system, the nervous

system, and the immune system (Contrada and

Baum, 2011). The biological response to stress is

differentiated between acute reaction and chronic

reaction. Acute reaction triggers a rapid release of

noradrenaline and adrenaline through the

sympathetic-medullary-adrenal axis (SMA Axis).

While the chronic reaction activating the

hypothalamic-pituitary-adrenal (HPA Axis) produces

cortisol (Matteri et al., 2000).

Cortisol is a glucocorticoid hormone produced

by the adrenal glands and synthesized from

cholesterol (Silverthorn, 2007). Cortisol levels are

regulated by adrenocorticotropic hormone (ACTH),

which response to corticotropin-releasing hormone

(CRH). Cortisol has widespread action such as

reduces inflammation, suppresses the immune

system, helps the body to manage stress, and

increases blood sugar through gluconeogenesis

(Guyton and Hall, 2016). Serum cortisol levels is

often used as an indicator of stress conditions

(Möstl and Palme, 2002). Forced swim test can be

used as a stressor which stimulate the release of

cortisol hormone (Khaleel Jameel et al., 2014).

Essential oils are a volatile product of a plant that

Effect of 10 .

DOI: 10.5220/0009954500130017

In Proceedings of the 2nd International Conference of Essential Oils (ICEO 2019), pages 13-17

ISBN: 978-989-758-456-5

has a scent so it is often used for cosmetic products,

perfumes, and aromatherapy. Essential oils obtained

from plants by steam distillation. One of the essential

oils that can be used is lavender. Lavender has more

than 30 species, dozens of subspecies, and hundreds

of hybrids, some of which are Lavandula angustifolia

(English Lavender), Lavandula stoechas (French

Lavender), Lavandula latifolia, and Lavandula

intermedia (Cavanagh and Wilkinson, 2002).

Lavandula angustifolia derived from the

Lamiaceae family is a native Mediterranean plant and

thrives in highland areas (Verma et al., 2010).

Lavender has two major constituents, there are

linalool and linalyl acetate (Kıvrak, 2018). Lavender

essential oil has many benefits such as relieve pain,

improve sleep quality, antimicrobial, anxiolytic, and

repellent (Cavanagh and Wilkinson, 2002).

The administration of lavender essential oil can be

oral, inhalation, and topical (Dornic et al., 2016).

Research showed the use of lavender essential oil in

inhalation can decrease the cortisol levels in blood

and saliva (Kim et al., 2012; Hosseini et al., 2016).

Moreover, oral administration of lavender essential

oil has an anxiolytic effect (Kasper et al., 2010). The

study of Jager et al. (1992) suggested that linalool

and linalyl acetate (active components of lavender

essential oil) are rapidly absorbed through the skin

and reaching peak levels after 19 minutes (Jager et

al., 1992).

This study aims to research the usage of natural

topical medication like lavender essential oil balm to

investigate the effectiveness for stress conditions.

Currently, treatment for stress conditions always

oriented to synthetic drugs such as psychotropic

drugs. These drugs are susceptible to misuse. Serum

cortisol levels are the indicator of stress conditions.

2 METHODS

2.1 Animals

Male Wistar rats (2-3 months, 100-200 grams) were

habituated for 7 days before the experiment. Foods

were given 60 grams and water was available ad

libitum. Animals were individually housed in the

cage.

2.2 Materials

Placebo balm consists of 1-gram beeswax and 5

ml (4.62 gram) virgin coconut oil as vehicle formula

and no active ingredient formula was given. 10%

lavender essential oil balm consists of 1-gram

beeswax and 5 ml (4.62 gram) virgin coconut oil as

vehicle formula and 0.625-gram lavender essential oil

(Lavandula angustifolia) as active formula.

2.3 Experimental Procedures

This study used 36 male Wistar rats divided into 4

groups (negative control = no stressor; positive

control = stressor only; placebo = stressor + placebo;

= stressor + Lavender 10%), the forced swim test

was given as the stressor. The cylinder was filled with

lukewarm water to a height of 30 cm. Animal was

placed in a water-filled cylinder for 10 second every

day for 30 days. Lavender essential oil balm was

given to the treatment group (L

) on the back (was

shaved 2x2 cm) within 30 minutes after the forced

swim test. These experiments were conducted

between 6:00 AM and 8:00 AM.

All of the rats were sacrificed on the 30th day.

The blood was collected via intracardiac puncture

and was centrifuged for plasma collection. All

plasma was stored in the freezer until assayed by

ELISA.

2.4 Statistic

This study This study was an experimental study

with a post-test only control group design. All the

results are presented as means ± standard error of

the mean. Statistical significance was analysed using

the Kruskal-Wallis test with the Mann-Whitney U as

post-hoc analysis by SPSS 25.0 software. P<0.05 was

considered to indicate a statistically significant

difference.

3 RESULT

Mann-Whitney U test showed that 10% lavender

essential oil balm significantly decreased the serum

cortisol levels in male Wistar rats compared to the

negative control group. Mean ± SD of serum

cortisol levels in 10% lavender essential oil balm

and negative control group were 684.19 ± 54.081

and 712.95 ± 129.589 (Table 1).

Also 10% lavender essential oil balm significantly

decreased the serum cortisol levels in male Wistar

rats compared to untreated rats. Mean ± SD of serum

cortisol levels in 10% lavender essential oil balm and

positive control group were 684.19 ± 54.081 and

728.13 ± 48.125 (Table 2).

Results showed no significant difference in

serum cortisol levels in the placebo balm group and

positive control group. Mean ± SD of serum cortisol

levels in the placebo balm group and positive

control group were 699.59 ± 64.135 and 728.13 ±

48.125 (Table 3).

Table 1: Comparison of negative control group and

treatment group.

Parameter

Sig

(p<0.05)

Serum Cortisol

(mean ± SD)

712.95 ±

129.589

684.19 ±

54.081

0.007*

: no stressor and no treatment were given

: stressor and 10% lavender essential oil balm

were given

p<0.05* Statistically significant

Table 2: Comparison of positi ve contro l gro up

an d treatment group

Parameter

Sig

(p<0.05)

Serum Cortisol

(mean ± SD)

728.13 ±

48.125

684.19 ±

54.081

0.041*

: stressor and no treatment were given

: stressor and 10% lavender essential oil balm

were given

p<0.05* Statistically significant

Table 3: Comparison of positive control gr o up

an d placebo group

Parameter

Sig

(p<0.05)

Serum Cortisol

(mean ± SD)

728.13 ±

48.125

699.59 ±

64.135

0.07

: stressor and no treatment were given

P: stressor and placebo balm were given

p>0.05 Statistically not significant

Table 4: Serum cortisol levels in

Groups

Serum cortisol

Negative control

712.95 ±

129.589

Positive control

728.13 ±

48.125

Placebo

699.59 ±

64.135

Lavender 10%

684.19 ±

54.081

4 DISCUSSION

The present study was planned to evaluate the effect

of 10% lavender essential oil balm on serum cortisol

levels of forced swim test models inducing stress in

male Wistar rats. As shown in Table 1 and Table 2,

serum cortisol levels were significantly lower after

given 10% lavender essential oil balm. These results

were similar to the previous study.

Lee and Cho (2014) studied that both lavender

essential oil inhalation and rosemary essential oil

inhalation reduced saliva cortisol levels in twenty

healthy South Korean students. However, the

lavender group reduced the saliva cortisol

levels more than the rosemary group.

Furthermore, lavender essential oil also

significantly decreased stress index and mood

index in subjects (Lee and Cho, 2014). In another

study, four weeks of inhalation of lavender

essential oil could reduce the level of saliva

cortisol and daytime blood pressure in

prehypertensive and hypertensive subjects, (Kim et

al., 2012). Hosseini et al. (2016) also reported that

inhalation of lavender essential oil showed

anxiolytic effect that decreased serum cortisol

levels in candidates for open-heart surgery

(Hosseini et al., 2016).

In this study, the researcher also examined the

effects of placebo balm against the serum cortisol

levels in male Wistar rats. We showed that no

significant differences were observed in serum

cortisol levels between the placebo balm and the

control group (Table 3). These results indicated that

the constituent of placebo balm did not reduce

serum cortisol levels.

Lavender essential oil has two major

constituents: linalool and linalyl acetate (Verma et

al., 2010; Kıvrak, 2018). A study was done by

Umezu et al. (2006) observed that male mice given

lavender essential oil intraperitoneally displayed an

anti-conflict effect in the Geller test and Vogel test.

Furthermore, the major constituents of lavender

essential oil (linalool and linalyl acetate) were also

evaluated and the only linalool showed

anxiolytic effect ( Umezu et al., 2006). Souto-

Maior et al. (2011) reported an anxiolytic effect for

inhaled linalool on male mice using the elevated

plus-maze test and light/dark box test. Also, inhaled

linalool did not appear to cause muscle relaxation

or motor coordination because no significant

decreased time spent on the rotarod test (Souto-

Maior et al., 2011). However, Takahashi et al.

(2011) concluded that linalyl acetate and linalool

act synergistically to induce anxiolytic effect in the

elevated plus-maze test (Takahashi et al., 2011).

The mechanism for serum cortisol levels

reduction by 10% lavender essential oil balm

remains to be determined. These effects may occur

through GABA

A receptor

modulation by linalool

which increases brain GABA levels (Milanos et al.,

2017). GABA is the major inhibitory

neurotransmission in the brain. GABA suppresses

Effect of 10

the activity of the HPA Axis by inhibiting

paraventricular nuclei in the hypothalamus

(Cullinan et al., 2008). Therefore, impaired

secretion of corticotropin- releasing hormone

(CRH) by paraventricular nuclei will disturb the

secretion of adrenocorticotropic hormone by the

anterior pituitary gland (Hannibal and Bishop,

2014). As a result of HPA Axis suppression, the

cortisol levels secretion by the adrenal gland will be

reduced.

5 CONCLUSIONS

This study was performed to investigate the

effectiveness of 10% lavender essential oil balm for

stress conditions. These result demonstrate that 10%

lavender essential oil balm reduced the serum cortisol

levels in male Wistar rats. The 10% lavender essential

oil balm determined to have a meaningful anxiolytic

effect. These results indicate that lavender essential

oil balm can be used as an alternative treatment to

relieve stress but should be further researched for

other biochemical parameters.

REFERENCES

Cavanagh, H, M, A., Wilkinson, J, M., 2002. Biological

Activities of Lavender Essential Oil, Phytotherapy

Research, 16(4), 301–308.

Contrada, R, J., Baum, A., 2011. The Handbook of Stress

Science: Biology, Psychology, and Health. New

York, Springer Pub.

Cullinan, W, E., Ziegler, D, R., Herman, J, P., 2008.

Functional Role of Local Gabaergic Influences on the

HPA Axis. Brain Structure and Function, 213(1–2),

63–72.

Dornic, N., Ficheux, A, S., Roudot, A, C., Saboureau, D.,

Ezzedine, K., 2016. Usage Patterns of Aromatherapy

among the French General Population: A Descriptive

Study Focusing on Dermal Exposure. Regulatory

Toxicology and Pharmacology, 76, 87–93.

Fink, G., 2016. Stress, Definitions, Mechanisms, and

Effects Outlined Lessons from Anxiety, Stress Concepts

and Cognition, Emotion, and Behavior. Elsevier Inc.

Gradus, J, L., Qin, P., Lincoln, A, K., Miller, M., Lawler, E.

V., Sorensen, H, T., Lash, T, L., 2010. Acute Stress

Reaction and Completed Suicide. International

Journal of Epidemiology, 39(6), 1478–1484.

Guyton, A, C., Hall, J, E., 2016. Textbook of Medical

Physiology 13th Edition. Philadelphia, PA: Elsevier.

Hannibal, K, E., Bishop, M, D., 2014. Chronic Stress,

Cortisol Dysfunction, and Pain: A

Psychoneuroendocrine Rationale for Stress

Management in Pain Rehabilitation. Physical Therapy,

94(12), 1816–1825.

Hosseini, S., Heydari, A., Vakili, M, A.,

Moghadam, S., Tazyky, S., 2016. Effect of

Lavender Essence Inhalation on the Level of Anxiety

and Blood Cortisol in Candidates for Open-Heart

Surgery. Iranian Journal of Nursing and Midwifery

Research, 21(4), 397.

Jager, W., Bu ch bau er , G. , J i r o vet z, L. ,

Fritze r , M . , 1992. Percutaneous Absorption of

Lavender Oil from a Massage Oil. Journal of the

Society of Cosmetic Chemists, 43(1), 49–54.

Kasper, S., Gastpar, M., Muller, W, E., Volz, H, P.,

Moller, H, J., Dienel, A., Schlafke, S., 2010. Silexan,

an Orally Administered Lavandula Oil Preparation,

is Effective in the Treatment of “Subsyndromal”

Anxiety Disorder: A Randomized, Double-Blind,

Placebo Controlled Trial. International Clinical

Psychopharmacology, 25(5), 277–287.

Kementerian Kesehatan. 2013. Riset Kesehatan Dasar

(Riskesdas) 2013. Jakarta, Badan Penelitian dan

Pengembangan Kesehatan, Kementerian Kesehatan

Republik Indonesia.

Kementerian Kesehatan. 2018. Riset Kesehatan Dasar

(Riskesdas) 2018. Jakarta, Badan Penelitian dan

Pengembangan Kesehatan, Kementerian Kesehatan

Republik Indonesia.

Khaleel Jameel, M., Joshi, A, R., Dawane, J, S., Padwal, M.,

Joshi, A., Pandit, V., Melinkeri, R., 2014. Effect of

Various Physical Stress Models on Serum Cortisol

Level in Wistar Rats. Journal of Clinical and

Diagnostic Research, 8(3), 181–183.

Kim, I., Kim, C., Seong, K., Hur, M, H., Lim, H, M.,

Lee, M, S., 2012. Essential Oil Inhalation on Blood

Pressure and Salivary Cortisol Levels in

Prehypertensive and Hypertensive Subjects.

Evidence-Based Complementary and Alternative

Medicine, 2012, 1–9.

Kıvrak, Ş., 2018. Essential Oil Composition and

Antioxidant Activities of Eight Cultivars of Lavender

and Lavandin from Western Anatolia. Industrial

Crops and Products, 117, 88–96.

Lee, A., Cho, H., 2014. The Effects Caused by Lavender

and Rosemary for Salivary Cortisol, Stress Levels

and Mood Alteration. Fashion business, 17(6), 18–

27.

Matteri, R, L., Carroll, J, A., Dyer, C, J., 2000.

Neuroendocrine Responses to Stress. The biology of

animal stress: basic principles and implications for

animal welfare, 43–62.

Milanos, S., Elsharif, S., Janzen, D., Buettner, A.,

Villmann, C., 2017. Metabolic Products of Linalool

and Modulation of GABAA Receptors. Frontiers in

Chemistry, 5, 1–9.

Möstl, E., Palme, R., 2002. Hormones as Indicators of

Stress. Domestic Animal Endocrinology, 23(1–2), 67–

74.

Silverthorn, D, U., 2007 Human physiology : An

Integrated Approach. San Francisco,

Pearson/Benjamin Cummings.

Souto-Maior, F, N., de Carvalho, F, L., de Morais,

L, C, S, L., Netto, S, M., de Sousa, D, P., de

Almeida, R, N., 2011. Anxiolytic-Like Effects of

Inhaled Linalool Oxide in Experimental Mouse

Anxiety Models. Pharmacology Biochemistry and

Behavior, 100(2), 259–263.

Takahashi, M., Satou, T., Ohashi, M., Hayashi, S.,

Sadamoto, K., Koike, K., 2011. Interspecies

Comparison of Chemical Composition and Anxiolytic-

Like Effects of Lavender Oils upon Inhalation. Natural

Product Communications, 6(11), 1769–74.

Umezu, T., N a g an o , K . , I t o , H., K o s a kai,

K. , Sak a n i wa, M . , M o rt a , M. , 2006.

Anticonflict Effects of Lavender Oil and

Identification of Its Active Constituents.

Pharmacology Biochemistry and Behavior, 85(4),

713–721.

Verma, R. S., Rahman, L, U., Chanotiya, C, S., Rk, V.,

Chauhan, A., Singh, A., Yadav, A, L., 2010. Esential

Oil Composition of Lavandula angustifolia Mill.

Cultivated in the Mid Hills of Uttarakhand, India.

Journal of the Serbian Chemical Society, 75(3), 343–

348.

Wada, K., Sairenchi, T., Haruyama, Y., Taneichi, H.,

Ishikawa, Y., Muto, T., 2013. Relationship between

the Onset of Depression and Stress Response

Measured by the Brief Job Stress Questionnaire among

Japanese Employees : A Cohort Study. 8(2).

World Health Organization. 2017. Depression and Other

Common Mental Disorders. Cc By-Nc-Sa 3.0 Igo, (1),

1–22.

Effect of 10