This Article

Citations


Creative Commons License
Except where otherwise noted, this work is licensed under Creative Commons Attribution-NonCommercial 4.0 International License.

Anxiolytic Effect of Borago officinalis (Boraginaceae) Extract in Male Rats


1 Neurophysiology Research Center, Hamadan University of Medical Sciences, Hamadan, IR Iran
*Corresponding author: Alireza Komaki, Neurophysiology Research Center, Hamadan University of Medical Sciences, Hamadan, IR Iran. Tel: +98-8138380267, Fax: +98-8138380131, E-mail: alirezakomaki@gmail.com.
Avicenna Journal of Neuro Psych Physiology. 2015 February; 2(1): e27189 , DOI: 10.17795/ajnpp-27189
Article Type: Research Article; Received: Jan 1, 2015; Revised: Feb 1, 2015; Accepted: Feb 4, 2015; epub: Feb 20, 2015; ppub: Feb 28, 2015

Abstract


Background: Medicinal plants with natural antioxidants have been shown to be beneficial in a variety of complications such as anxiety. The elevated plus-maze (EPM) is one of the most widely used models to assess anxiety in small rodents.

Objectives: This study was designed to characterize the anxiolytic-like activity of Borago officinalis (Linnaeus, family Boraginaceae) or Borage flowers extract, using an EPM test.

Materials and Methods: Male Wistar rats weighing 220-250 grams were used in the present study. Thirty minutes after an intraperitoneal (IP) injection of the Borage extract (50, 100, 200 mg/kg) or saline, each animal was placed in the EPM. Animal behaviors in the experimental sessions were recorded by a video camera located above the maze, interfaced with a monitor and a computer in an adjacent room. The time spent in the open arms, the percentage of entries into the open arms of the EPM and the numbers of entries into the closed arms were recorded for five minutes.

Results: Statistical analysis indicated that acute IP injection of Borage extract before an EPM trial significantly increased the time spent in open arms and percentage of open arms entries. Whereas, the extract had no effect on the number of closed arm entries.

Conclusions: Our results demonstrated that injection of Borage extract might have an anxiolytic profile in rats. However, the exact mechanism (s) related to the active compound (s) in Borage extract should be elucidated in future studies.

Keywords: Borago; Rat; Medicinal Plants; Antioxidants

1. Background


Medicinal plants have been used from ancient time for their medicinal values as well as to impart flavor to food (1). The use of plants in the management of illnesses has been continuously increased over time as they are readily and cheaply available healthcare alternatives (2). Nowadays, the crude extracts and dry powder samples from medicinal and aromatic plants and their species have been used for the development and preparation of alternative traditional medicine and food additives (3, 4). Various types of herbal medicines have been used as anxiolytics in different parts of the world (5). Borago officinalis (Linnaeus, family Boraginaceae) or Borage is an annual herb with nutritional value used in traditional medicine and culinary uses in some countries (6, 7). Borage is cultivated around the world but is native to Europe, North Africa and Asia (8, 9). This plant was investigated from some other points of view, mostly regarding their medicinal properties (10). Borage products, mainly seeds oil, flowers and leaves are used for medicinal and culinary purposes (11). The flowers are blue and rarely appear white or rose colored (12). In Iran, borage blooms during April until September (11). This plant is harvested in flowering period before the start of seed formation (13-15). It has been reported that borage has antipyretic, aphrodisiac, antispasmodic, demulcent, antihypertensive and diuretic effects. Furthermore, the products of Borage are used for the treatment of bronchitis, palpitations, cramps and diarrhea (16-18). This plant can be also considered useful for the treatment of diseases such as premenstrual pain, multiple sclerosis, rheumatoid arthritis and asthma (19-21). There is a variety of animal tests for the investigation of anxiolytic effects of substances (22). One of the most widely used models for detecting both anxiolytic- and anxiogenic-like effects of agents in small rodents is elevated plus-maze test (EPM) (23, 24). In this animal model, the increase percentage of entries into the open arms compared to the total entries reveals an anxiolytic effect of substances. An increase of the time and proportion of the entrances into the open arms without a changed locomotor activity is regarded as a powerful marker of anxiolytic substance effect (25-28).

2. Objectives


Medicinal plants with natural antioxidants have been shown to be beneficial in a variety of complications such as anxiety (29). Furthermore, increasing evidence suggests that impairment of antioxidant defense is important in the process of emotional disorders, such as depression and anxiety (30). On the other hand, it has been reported that borage extract constitutes a cheap source of antioxidant activity (11, 31-33). However, there are no published reports in literature about the effect of Borago officinalis extract on anxiety. Therefore, this study was designed to characterize the anxiolytic-like activity of Borago officinalis flowers extract, using an EPM test.

3. Materials and Methods


3.1. Preparation of Plant Extract

Borage flowers were collected from Hamadan region (Iran) in the spring and identified at the Botanic Institute of Hamadan University of Medical Sciences. The plant flowers were separated, cleaned, washed, homogenized and then dried for storage. Then, dried flowers were ground and produced a crushed powder (19). The powdered material was soaked into 70% ethanol. The extract was then taken to the laboratory for the process of evaporation. The evaporation process involved total removal of ethanol and water with which the extraction took place from the extract. The extracts of flowers were dried at 40ºC and finally freeze-dried (34). The doses of aqueous suspensions of Borage flowers extract are expressed as milligrams of dried extract per kilograms of rat body weight (mg/kg). The extracts were redissolved in their solvents before each individual experiment (35).

3.2. Animals

Male Wistar rats weighing 220-250 grams were transported from animal house to a room adjacent to the test laboratory 72 hours before the test. They were housed under a controlled condition; 12:12 dark/ light cycle (lights on at 08:00 AM) at 22 ± 2ºC. Animals had free access to food and water. Rats were randomly assigned to different treatment groups (n = 10). Animals were divided into four groups: control group and Borage extract groups [50, 100, 200 mg/kg intraperitoneal (IP)]. Animals were tested under the same experimental conditions. All animals received humane care according to the criteria outlined in the Guide for the Care and Use of Laboratory Animals prepared by the National Academy of Sciences and published by the National Institutes of Health (NIH publication 86-23 revised 1985) and the study was also approved by the local ethics committee of Hamadan University of Medical Sciences.

3.3. Elevated Plus-Maze Test

EPM design was similar to that originally described by Lister (36). In brief, the apparatus was composed of two open (50 cm × 10cm × 1 cm) and two enclosed (50 cm × 10 cm × 50 cm) arms with a central platform (10 cm × 10 cm) to form a plus sign. The maze was elevated 50 cm above the floor. Thirty minutes after an IP injection of the Borage extract (50, 100, 200 mg/kg) or saline, animals behavior in the percentage of entries into the arms and time spent in each arms were recorded during five minutes (37). Animal behaviors in the experimental sessions were recorded by a video camera located above the maze, interfaced with a monitor and a computer in an adjacent room. The open-arm entries and open-arm time were used as indices of anxiety and the number of entries into the closed arms reduction of spontaneous locomotion in rat. After the test, the maze was carefully cleaned with a wet tissue paper (10% ethanol solution).

3.4. Statistical Analysis

Calculation of the total time spent in each of the arms, number of entries into the closed arms of EPM and percentage of entries into the each arms compared to total entries were performed using computerized analysis. The difference between the groups was determined by one-way Analysis of Variance (ANOVA) followed by Tukey post hoc test. In all cases differences were considered significant if P < 0.05. Results were expressed as Mean ± SEM.

4. Results


One-way ANOVA indicated that compared to the control group, extract of Borago officinalis groups spent more time in the open arms. Tukey post hoc test analysis showed that extract-treated group spent more time in the open arms in the doses of 100 and 200 mg/kg (P < 0.05), whereas effect of 50 mg/kg was not significant compared with the control group. The effects of different doses of Borago officinalis extract on the duration of time spent in the open arms are shown in Figure 1. One-way ANOVA indicated that compared to the control group, acute doses of Borago officinalis exerted significant effects on the percentage of entries into the open arms (Figure 2). A Tukey’s post hoc analysis revealed significant effects of Borago officinalis treatment on the percentage of entries into the open arms (50 mg/kg; P < 0.05 and 100 and 200 mg/kg; P < 0.01). The number of entries into the closed arms was not significantly different between the Borago officinalis treated and control groups (Figure 3).

Figure 1.
The Effects of Borage Extract (50, 100, 200 mg/kg) on the Time Spent in the Open and Closed Arms of the EPM Within the Five-Minute Test Period. Rats Were 10 in the Treated Groups
Figure 2.
The Effects of Borage Extract (50, 100, 200 mg/kg) on the Percentage of Entries Into the Open and Closed Arms of the EPM Within the Five-Minute Test Period
Figure 3.
The Effects of Borage Extract (50, 100, 200 mg/kg) on the Number of Entries Into the Closed Arms of the EPM During the Five-Minute Test Session

5. Discussion


Borage is a medicinal plant with different usages in pharmaceutical, industrial and forage fields used in production of drinks and salads (7). Many natural substances and plants exert well accepted medicinal effects on animal behaviors and are able to act on chronic conditions such as anxiety that do not respond well to current therapeutic treatments (38). Substances and drugs derived from traditional plants may have possible therapeutic correlation in the remedy of disorders such as anxiety (5). Based on the above topics, the purpose of this study was to distinguish anxiolytic-like activity of the extract prepared from Borago officinalis. The results of present study demonstrated that the extract of Borago officinalis increased both the percentage of the time spent in the open arms of the maze and percentage of entries into the open arms of the maze. In the other word, the extract was able to produce anxiolytic effect in rats. An increase in the percentage of entrances into the open arms and time spent in open arms, lacking a changed locomotors activity are confirmed as a potent sign for an anxiolytic compounds effect (23). Borage is of great interest among medical and nutritional research groups due to its high content of some useful compounds (10, 11, 39). Linolenic acid and palmitic acid are collected from flowers and high levels of α-Linolenic acid is in mature leaves (40). Borage oil has high content of γ-linolenic acid (33, 39). Borage oil has been reported to lower serum cholesterol, phospholipids and triglyceride levels (41). Borage oil is considered to improve task performance through attenuation of blood pressure, heart rate and temperature elevation in response to stress (42). Due to its folkloric use as an antispasmodic remedy and based on the traditional use of Borago officinalis in asthma, Borago officinalis extract was tested for its possible spasmolytic and bronchodilator effects (17). Borago officinalis seed oil is used in pharmaceutical products, especially for its high content in γ-linolenic acid, an essential and unusual fatty acid (10, 43). It has been shown that palmitic, linoleic and stearic acids were dominant in the seed oil of borage (44). Wettasinghe and Shahidi investigated the antioxidant and reactive oxygen species (ROS) scavenging properties of borage meal extract (45, 46). Extensive research has been conducted to reveal multiple neural substrates and mechanisms that contribute to the etiology of depression and anxiety, among which the imbalance between oxidation and antioxidant defense system has gained attention (30). Some studies demonstrated the role of oxidative stress in anxiety of rodents (47, 48). According to this, it has been shown that induction of oxidative stress in mice occurs in same time with anxiety (49). Diets including high levels of sugar were reported to increase the oxidation of proteins in brain and to cause anxiety in animals. Increased anxiety has been positively correlated with increased ROS levels. Interestingly, the induction of oxidative stress by a non-pharmacological method leads to anxiety like behaviors in rats (47). Among natural antioxidants, phenolic antioxidants are extensively available in plants (50-52). Effectiveness of antioxidant activity of plant extracts in vitro is probably caused by their ability to act as free radical scavengers (53, 54). It has been demonstrated that administration of Borage was protective against Aβ-induced memory and antioxidant deficit (6). Moreover, different works remarked the antioxidant activity of Borage extracts, especially related to their content in phenolic compounds (11, 46, 55). The use of Borage has become more popular, because its oil is one of the sources of γ-linolenic acid (15, 56-58). Moreover, several studies evaluated the associations between antioxidant activity of Borage extract and its γ-linolenic acid content (31, 32). Borage increased the antioxidant power of homogenate samples of hippocampus (46, 59). The protective effect of Borage on memory can be related to its function of scavenging free radicals and high content of γ-linolenic acid (39). In conclusion, our results demonstrated that injection of Borage extract might have an anxiolytic profile in rat. The presence of polyphenols, flavonoids and substances such as γ-linolenic acid in Borage flowers extract reinforces the anxiolytic effects of this plant found in this study. However, the exact mechanism(s) related to the active compound (s) in Borage extract should be elucidated in future studies.

Acknowledgments

The authors would like to express their gratitude to the staff of the Neurophysiology Research Center for helping them perform this project.

Footnotes

Authors’ Contributions: Study concept and design: Alireza Komaki, Bahman Rasouli and Siamak Shahidi. Acquisition of data: Bahman Rasouli and Alireza Komaki. Analysis and interpretation of data: Alireza Komaki and Bahman Rasouli. Drafting of the manuscript: Alireza Komaki and Siamak Shahidi. Critical revision of the manuscript for important intellectual content: Alireza Komaki, Bahman Rasouli. Statistical analysis: Alireza Komaki and Siamak Shahidi. Administrative, technical and material supports: Alireza Komaki and Bahman Rasouli. Study supervision: Alireza Komaki and Siamak Shahidi.
Funding/Support: This research was supported by a grant (Grant number: 87003581416) from Hamadan University of Medical Sciences, Hamadan, IR Iran.

References


  • 1. Hossain MA, AL-Raqmi KA, AL-Mijizy ZH, Weli AM, Al-Riyami Q. Study of total phenol, flavonoids contents and phytochemical screening of various leaves crude extracts of locally grown Thymus vulgaris. Asian Pac J Trop Biomed. 2013;3(9):705-10. [DOI] [PubMed]
  • 2. Bussmann RW, Swartzinsky P, Worede A, Evangelista P. Plant use in Odo-Bulu and Demaro, Bale region, Ethiopia. J Ethnobiol Ethnomed. 2011;7:28. [DOI] [PubMed]
  • 3. Rota MC, Herrera A, Martínez RM, Sotomayor JA, Jordán MJ. Antimicrobial activity and chemical composition of Thymus vulgaris, Thymus zygis and Thymus hyemalis essential oils. Food Control. 2008;19(7):681-7. [DOI]
  • 4. Oke F, Aslim B, Ozturk S, Altundag S. Essential oil composition, antimicrobial and antioxidant activities of Satureja cuneifolia Ten. Food Chem. 2009;112(4):874-9. [DOI]
  • 5. Grundmann O, Nakajima J, Seo S, Butterweck V. Anti-anxiety effects of Apocynum venetum L. in the elevated plus maze test. J Ethnopharmacol. 2007;110(3):406-11. [DOI] [PubMed]
  • 6. Ghahremanitamadon F, Shahidi S, Zargooshnia S, Nikkhah A, Ranjbar A, Soleimani Asl S. Protective effects of Borago officinalis extract on amyloid beta-peptide(25-35)-induced memory impairment in male rats: a behavioral study. Biomed Res Int. 2014;2014:798535. [DOI] [PubMed]
  • 7. Asadi-Samani M, Bahmani M, Rafieian-Kopaei M. The chemical composition, botanical characteristic and biological activities of Borago officinalis: a review. Asian Pac J Trop Med. 2014;7S1:S22-8. [DOI] [PubMed]
  • 8. Peiretti PG, Palmegiano GB, Salamano G. Quality and fatty acid content of borage (Borago officinalis L.) during the growth cycle. Ital J Food Sci. 2004;16(2):177-84.
  • 9. Ghahreman A. Flora of Iran,Tehran: Research Institute of Forests and Rangelands 250p-col illus Fr, Pe, En Icones Maps Geog J. 1997;16(2)
  • 10. Mondello L, Beccaria M, Donato P, Cacciola F, Dugo G, Dugo P. Comprehensive two-dimensional liquid chromatography with evaporative light-scattering detection for the analysis of triacylglycerols in Borago officinalis. J Sep Sci. 2011;34(6):688-92. [DOI] [PubMed]
  • 11. Ciriano MG, Garcia-Herreros C, Larequi E, Valencia I, Ansorena D, Astiasaran I. Use of natural antioxidants from lyophilized water extracts of Borago officinalis in dry fermented sausages enriched in omega-3 PUFA. Meat Sci. 2009;83(2):271-7. [DOI] [PubMed]
  • 12. Zargari A. Iranian medicinal plants. Tehran: Tehran Univ Pub; 1997.
  • 13. Yang W, Sokhansanj S, Tang J, Winter P. Determination of thermal conductivity, specific heat and thermal diffusivity of borage seeds. Biosyst Eng. 2002;82:169-76. [DOI]
  • 14. Simpson MJA. Comparison of swathing and desiccation of borage (Borago officinalis) and estimation of optimum harvest stage. Annals of Applied Biology. 1993;123(1):105-8. [DOI]
  • 15. Mhamdi B, Aidi Wannes W, Sriti J, Jellali I, Ksouri R, Marzouk B. Effect of harvesting time on phenolic compounds and antiradical scavenging activity of Borago officinalis seed extracts. Industrial Crops and Products. 2010;31(1):e1-4. [DOI]
  • 16. Usmanghani K, Saeed A, Alam MT. Indusyunic medicine. Karachi: University of Karachi press; 1997. p. 254-5.
  • 17. Gilani AH, Bashir S, Khan AU. Pharmacological basis for the use of Borago officinalis in gastrointestinal, respiratory and cardiovascular disorders. J Ethnopharmacol. 2007;114(3):393-9. [DOI] [PubMed]
  • 18. Duke JA. Handbook of medicinal herbs. CRC press; 2010.
  • 19. Ramandi NF, Najafi NM, Raofie F, Ghasemi E. Central composite design for the optimization of supercritical carbon dioxide fluid extraction of fatty acids from Borago officinalis L. flower. J Food Sci. 2011;76(9):C1262-6. [DOI] [PubMed]
  • 20. Senanayake SPJN, Shahidi F. Lipase-catalyzed incorporation of docosahexaenoic acid (DHA) into borage oil: optimization using response surface methodology. Food Chem. 2002;77(1):115-23. [DOI]
  • 21. Kim HR, Hou CT, Lee KT, Kim BH, Kim IH. Enzymatic synthesis of structured lipids using a novel cold-active lipase from Pichia lynferdii NRRL Y-7723. Food chem. 2010;122(3):846-9. [DOI]
  • 22. Sciolino NR, Smith JM, Stranahan AM, Freeman KG, Edwards GL, Weinshenker D, et al. Galanin mediates features of neural and behavioral stress resilience afforded by exercise. Neuropharmacology. 2015;89:255-64. [DOI] [PubMed]
  • 23. Skelly MJ, Weiner JL. Chronic treatment with prazosin or duloxetine lessens concurrent anxiety-like behavior and alcohol intake: evidence of disrupted noradrenergic signaling in anxiety-related alcohol use. Brain Behav. 2014;4(4):468-83. [DOI] [PubMed]
  • 24. Kochenborger L, Levone BR, da Silva ES, Taschetto AP, Terenzi MG, Paschoalini MA, et al. The microinjection of a cannabinoid agonist into the accumbens shell induces anxiogenesis in the elevated plus-maze. Pharmacol Biochem Behav. 2014;124:160-6. [DOI] [PubMed]
  • 25. Pellow S, Chopin P, File SE, Briley M. Validation of open:closed arm entries in an elevated plus-maze as a measure of anxiety in the rat. J Neurosci Methods. 1985;14(3):149-67. [PubMed]
  • 26. Komaki A, Nasab ZK, Shahidi S, Sarihi A, Salehi I, Ghaderi A. Anxiolytic Effects of Acute Injection of Hydro-Alcoholic Extract of Lettuce in the Elevated Plus-Maze Task in Rats. Avicenna J Neuro Psych Physio. 2014;1(1):e18695
  • 27. Komaki A, Haghgooyan A, Shahidi S, Sarihi A, Salehi I. Interaction Between L-Type Calcium Channels and Antagonist of Cannabinoid System on Anxiety in Male Rat. Avicenna J Neuro Psych Physio. 2014;1(2):e24450
  • 28. Komaki A, Abdollahzadeh F, Sarihi A, Shahidi S, Salehi I. Interaction between Antagonist of Cannabinoid Receptor and Antagonist of Adrenergic Receptor on Anxiety in Male Rat. Basic Clin Neurosci. 2014;5(3):218-24. [PubMed]
  • 29. Salim S, Sarraj N, Taneja M, Saha K, Tejada-Simon MV, Chugh G. Moderate treadmill exercise prevents oxidative stress-induced anxiety-like behavior in rats. Behav Brain Res. 2010;208(2):545-52. [DOI] [PubMed]
  • 30. Ding L, Zhang C, Masood A, Li J, Sun J, Nadeem A, et al. Protective effects of phosphodiesterase 2 inhibitor on depression- and anxiety-like behaviors: involvement of antioxidant and anti-apoptotic mechanisms. Behav Brain Res. 2014;268:150-8. [DOI] [PubMed]
  • 31. Rio-Celestino M, Font R, de Haro-Bailon A. Distribution of fatty acids in edible organs and seed fractions of borage (Borago officinalis L.). J Sci Food Agric. 2008;88(2):248-55. [DOI]
  • 32. del Rio M, Alcaide B, Rapoport H, Cabrera A, De Haro A, editor(s). Characterisation and evaluation of species of the Boraginaceae family as source of gamma-linolenic acid for Mediterranean conditions. XXVI International Horticultural Congress: The Future for Medicinal and Aromatic Plants 629; 2002; ISHS; p. 231-7.
  • 33. Bandoniene D, Murkovic M. The detection of radical scavenging compounds in crude extract of borage (Borago officinalis L.) by using an on-line HPLC-DPPH method. J Biochem Biophys Methods. 2002;53(1-3):45-9. [PubMed]
  • 34. Silva LF, Lima ES, Vasconcellos MC, Aranha ES, Costa DS, Mustafa EV, et al. In vitro and in vivo antimalarial activity and cytotoxicity of extracts, fractions and a substance isolated from the Amazonian plant Tachia grandiflora (Gentianaceae). Mem Inst Oswaldo Cruz. 2013;108(4):501-7. [DOI] [PubMed]
  • 35. Local Food-Nutraceuticals C. Understanding local Mediterranean diets: a multidisciplinary pharmacological and ethnobotanical approach. Pharmacol Res. 2005;52(4):353-66. [DOI] [PubMed]
  • 36. Lister RG. The use of a plus-maze to measure anxiety in the mouse. Psychopharmacology (Berl). 1987;92(2):180-5. [PubMed]
  • 37. Pellow S, File SE. Anxiolytic and anxiogenic drug effects on exploratory activity in an elevated plus-maze: a novel test of anxiety in the rat. Pharmacol Biochem Behav. 1986;24(3):525-9. [PubMed]
  • 38. Blanco MM, Costa CA, Freire AO, Santos JJ, Costa M. Neurobehavioral effect of essential oil of Cymbopogon citratus in mice. Phytomedicine. 2009;16(2-3):265-70. [DOI] [PubMed]
  • 39. Huang YS, Lin X, Redden PR, Horrobin DF. In vitro hydrolysis of natural and synthetic γ-linolenic acid-containing triacylglycerols by pancreatic lipase. J Am Oil Chem Soc. 1995;72(6):625-31. [DOI]
  • 40. Griffiths G, Brechany EY, Jackson FM, Christie WW, Stymne S, Stobart AK. Distribution and biosynthesis of stearidonic acid in leaves of Borago officinalis. Phytochem. 1996;43(2):381-6. [DOI]
  • 41. Gu JY, Wakizono Y, Dohi A, Nonaka M, Sugano M, Yamada K. Effect of dietary fats and sesamin on the lipid metabolism and immune function of Sprague-Dawley rats. Biosci Biotechnol Biochem. 1998;62(10):1917-24. [DOI] [PubMed]
  • 42. Mills DE, Prkachin KM, Harvey KA, Ward RP. Dietary fatty acid supplementation alters stress reactivity and performance in man. J Hum Hypertens. 1989;3(2):111-6. [PubMed]
  • 43. Griffiths G, Stobart AK, Stymne S. Delta 6- and delta 12-desaturase activities and phosphatidic acid formation in microsomal preparations from the developing cotyledons of common borage (Borago officinalis). Biochem J. 1988;252(3):641-7. [PubMed]
  • 44. Morteza E, Akbari GA, Moaveni P, Alahdadi I, Bihamta MR, Hasanloo T, et al. Compositions of the seed oil of the Borago officinalis from Iran. Nat Prod Res. 2014;1-4. [DOI] [PubMed]
  • 45. Wettasinghe M, Shahidi F. Scavenging of reactive-oxygen species and DPPH free radicals by extracts of borage and evening primrose meals. Food Chem. 2000;70(1):17-26. [DOI]
  • 46. Wettasinghe M, Shahidi F. Antioxidant and free radical-scavenging properties of ethanolic extracts of defatted borage (Borago officinalis L.) seeds. Food chem. 1999;67(4):399-414. [DOI]
  • 47. Vollert C, Zagaar M, Hovatta I, Taneja M, Vu A, Dao A, et al. Exercise prevents sleep deprivation-associated anxiety-like behavior in rats: potential role of oxidative stress mechanisms. Behav Brain Res. 2011;224(2):233-40. [DOI] [PubMed]
  • 48. de Almeida AA, de Carvalho RB, Silva OA, de Sousa DP, de Freitas RM. Potential antioxidant and anxiolytic effects of (+)-limonene epoxide in mice after marble-burying test. Pharmacol Biochem Behav. 2014;118:69-78. [DOI] [PubMed]
  • 49. Masood A, Nadeem A, Mustafa SJ, O'Donnell JM. Reversal of oxidative stress-induced anxiety by inhibition of phosphodiesterase-2 in mice. J Pharmacol Exp Ther. 2008;326(2):369-79. [DOI] [PubMed]
  • 50. Rafieian-Kopaei M, Baradaran A, Rafieian M. Oxidative stress and the paradoxical effects of antioxidants. J Res Med Sci. 2013;18(7):629. [PubMed]
  • 51. Kafash-Farkhad N, Asadi-Samani M, Rafieian-Kopaei M. A review on phytochemistry and pharmacological effects of Prangos ferulacea (L.) Lindl. Life Sci J. 2013;10(8s):360-7.
  • 52. Asadi-Samani M, Rafieian-Kopaei M, Azimi N. Gundelia: a systematic review of medicinal and molecular perspective. Pak J Biol Sci. 2013;16(21):1238-47. [PubMed]
  • 53. Yang UJ, Park TS, Shim SM. Protective effect of chlorophyllin and lycopene from water spinach extract on cytotoxicity and oxidative stress induced by heavy metals in human hepatoma cells. J Toxicol Environ Health A. 2013;76(23):1307-15. [DOI] [PubMed]
  • 54. Melidou M, Riganakos K, Galaris D. Protection against nuclear DNA damage offered by flavonoids in cells exposed to hydrogen peroxide: the role of iron chelation. Free Radic Biol Med. 2005;39(12):1591-600. [DOI] [PubMed]
  • 55. Wettasinghe M, Shahidi F, Amarowicz R, Abou-Zaid MM. Phenolic acids in defatted seeds of borage (Borago officinalis L.). Food Chem. 2001;75(1):49-56. [DOI]
  • 56. Venskutonis PR, Dauksas E, Sivik B. Use of immobilised lipase from Candida antarctica in supercritical fluid extraction of borage (Borago officinalis L.) seed oil. Food Technol Biotechnol . 2008;46(2):157-63.
  • 57. Soto C, Concha J, Zuniga ME. Antioxidant content of oil and defatted meal obtained from borage seeds by an enzymatic-aided cold pressing process. Process Biochemistry. 2008;43(6):696-9. [DOI]
  • 58. Lin TJ, Chen SW. Enrichment of n-3 polyunsaturated fatty acids into acylglycerols of borage oil via lipase-catalyzed reactions under supercritical conditions. Chemical Engineering Journal. 2008;141(1-3):318-26. [DOI]
  • 59. Segovia F, Lupo B, Peiró S, Gordon M, Almajano M. Extraction of Antioxidants from Borage (Borago officinalis L.) Leaves—Optimization by Response Surface Method and Application in Oil-in-Water Emulsions. Antioxidants. 2014;3(2):339-57. [DOI]

Figure 1.

The Effects of Borage Extract (50, 100, 200 mg/kg) on the Time Spent in the Open and Closed Arms of the EPM Within the Five-Minute Test Period. Rats Were 10 in the Treated Groups
Comparisons were made using a one-way anova followed by post hoc tukey test. *P < 0.05 and ** P < 0.01 vs. control.

Figure 2.

The Effects of Borage Extract (50, 100, 200 mg/kg) on the Percentage of Entries Into the Open and Closed Arms of the EPM Within the Five-Minute Test Period
Rats were 10 in the treated groups. Comparisons were made using a one-way ANOVA followed by post hoc Tukey test. * P < 0.05 and ** P < 0.01 vs. control.

Figure 3.

The Effects of Borage Extract (50, 100, 200 mg/kg) on the Number of Entries Into the Closed Arms of the EPM During the Five-Minute Test Session
Data represent Means ± SEM. Animals were 10 in the treated groups.