Evaluation of Ajuga bracteosa for antioxidant, anti-inflammatory, analgesic, antidepressant and anticoagulant activities.
BACKGROUND
Ajuga bracteosa has been extensively used traditionally for the treatment of a variety of diseases. The aim of the study was to scientifically validate the wide-scale exploitation of A. bracteosa in folk medicine various in vitro and in vivo assays. Moreover, these activities were related to the intrinsic biologically active phytoecdysteroids of A. bracteosa.


METHODS
Aerial and root parts of A. bracteosa were first extracted separately with chloroform (AbCA and AbCR) and the residue was again extracted with methanol (AbMA and AbMR). Total flavonoid and phenolic contents were assayed as quercetin (QE) and gallic acid equivalents (GAE), respectively. These extracts were analyzed for in vitro antioxidant assessment including DPPH and H2O2(% inhibition of free radicals), and reducing power and phosphomolybdenum methods (ascorbic acid equivalents AAE mg/g DW). Further, these extracts were assayed in vivo in separate groups of Sprague-Dawley rats for carrageenan induced rat paw edema inhibition, hotplate antinociception, forced swim antidepression and anticoagulation. Dose of each crude extract and standard drug given to rats was 200 mg/Kg- and 10 mg/10 mL/Kg body weight respectively. Plant extracts and standard drugs were administered orally, 60 min prior to the conduction of assays. Moreover, biologically active phytoecdysteroids were screened in A. bracteosa with the help of RP-HPLC.


RESULTS
AbMA represented highest values of flavonoids (QE 1.98 % DW) and phenolic contents (GAE 5.94 % DW), significantly scavenged DPPH radicles (IC5036.9) and reduced ferric ions with 718.4 mg ascorbic acid equivalent/g (AAE). Highest total antioxidant capacity was expressed by AbMR (927 mg AAE) with an IC50value 19.1 μg/mL. The extracts which were found potent anti-oxidants, were also good at in vivo activities. AbMA significantly reduced edema in all the three hours of treatment (67.9, 70.3 and 74.3 %). AbMA also showed maximum nociceptor suppression in analgesic assay by delaying the time to start licking of paws in rats (57.7 ± 4.9 s). In addition, maximum anti-coagulation was also exhibited by AbMA (89.3 s), while all extracts were found strong antidepressants (≤15.66 s immobility time). Screening of biologically active phytoecdysteroids revealed the presence of 20-hydroxyecdysone (20-HE), makisterone (MKA), cyasterone (CYP) and ajujalactone (AJL). Total phytoecdysteroid content found in A. bracteosa was 1232.5 μg/g DW and 20-HE was most abundant (1232.5 μg/g DW) as compared to other phytoecdysteroids.


CONCLUSIONS
Based on the tested in vitro and in vivo activities, AbMA was found to be a promising bioactive extract. These activities can be attributed to the intrinsic polyphenols and phytoecdysteroids contents of A. bracteosa.
Department of Botany, The University of Poonch, Rawalakot, Azad Jammu and Kashmir, 12350, Pakistan.
Waqas Khan
WK
Kayani
Department of Bioinformatics and Biosciences, Capital University of Science and Technology, Islamabad, Pakistan.
Erum
E
Dilshad
Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, 45320, Pakistan.
Tanveer
T
Ahmed
Department of Biochemistry and Molecular Biology, University of Gujrat, 50700, Gujrat, Pakistan.
Hammad
H
Ismail
Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, 45320, Pakistan. bushramirza@qau.edu.pk.
Bushra
B
Mirza
Ajuga bracteosa; Anti-inflammatory; Antidepressant; Antioxidants; Phytoecdysteroids; RP-HPLC
Curr Neuropharmacol. 2014 Mar;12(2):140-7
24669208
Ukr Biokhim Zh (1999). 2001 May-Jun;73(3):21-9
12035548
Eur J Pharmacol. 2009 Aug 15;616(1-3):128-33
19549518
Bull Exp Biol Med. 2008 Sep;146(3):328-33
19240852
Comp Biochem Physiol A Mol Integr Physiol. 2003 Sep;136(1):113-26
14527634
J Ethnobiol Ethnomed. 2011 May 30;7:15
21624132
Saudi Pharm J. 2013 Apr;21(2):143-52
24936134
Ukr Biokhim Zh (1978). 1992 Jan-Feb;64(1):114-7
1519340
Asian Pac J Trop Biomed. 2012 Mar;2(3):185-8
23569895
Am Fam Physician. 2005 Feb 1;71(3):483-90
15712623
CNS Drugs. 2012 Jun 1;26(6):477-90
22668245
J Ethnopharmacol. 2015 Jul 1;169:335-46
25952168
Indian J Med Res. 2013;138:99-103
24056562
J Insect Sci. 2003;3:7
15844229
J Ethnopharmacol. 2011 Apr 26;135(1):116-25
21382463
Chem Biodivers. 2007 Jan;4(1):72-83
17256736
J Ethnopharmacol. 2011 Jan 27;133(2):928-30
21073945
Ukr Biokhim Zh (1999). 1999 Nov-Dec;71(6):37-42
10820845
Food Chem Toxicol. 1999 Sep-Oct;37(9-10):949-62
10541450
Ukr Biokhim Zh (1999). 1999 May-Jun;71(3):35-8
10609320
Molecules. 2008 Nov 12;13(11):2804-13
19015620
Molecules. 2010 Jun 14;15(6):4324-33
20657444
Psychoneuroendocrinology. 2015 Jan;51:164-75
25462890
J Ethnopharmacol. 2003 Mar;85(1):19-23
12576198
Proc Soc Exp Biol Med. 1962 Dec;111:544-7
14001233
Osong Public Health Res Perspect. 2013 Oct;4(5):246-54
24298440
Eksp Klin Farmakol. 1997 Sep-Oct;60(5):58-60
9483409
Anal Biochem. 1999 May 1;269(2):337-41
10222007
J Dent. 2009 Jun;37(6):413-23
19303186
J Immunol. 2003 Sep 15;171(6):3010-8
12960326
Bioresour Technol. 2008 Jun;99(9):3365-71
17904358
Daru. 2014 Jul 14;22:56
25022284
BMC Complement Altern Med. 2015 Jun 27;15:199
26115918
Nature. 1977 Apr 21;266(5604):730-2
559941
Semin Hematol. 2002 Jul;39(3):145-57
12124677
J Cell Mol Med. 2010 Apr;14(4):840-60
19754673
Mutat Res. 2002 Aug 26;519(1-2):103-19
12160896
Oxid Med Cell Longev. 2009 Apr-Jun;2(2):63-7
20357926
Antioxidants (Basel). 2014 Oct 20;3(4):671-83
26785233
Fitoterapia. 2002 Aug;73(5):375-80
12165331
Fitoterapia. 2004 Mar;75(2):162-7
15030920
Free Radic Res. 1997 Nov;27(5):511-32
9518068
J Ethnopharmacol. 2004 Feb;90(2-3):205-15
15013182
Carcinogenesis. 1989 Jun;10(6):1003-8
2470525
Int J Immunopathol Pharmacol. 2013 Oct-Dec;26(4):847-54
24355219
Physiol Rev. 2002 Jan;82(1):47-95
11773609
J Food Sci. 2014 Feb;79(2):R129-37
24446991
J Pharmacol Exp Ther. 1997 Aug;282(2):967-76
9262365
Curr Med Chem. 2005;12(2):153-72
15638733
J Vis Exp. 2012 Jan 29;(59):e3638
22314943
Br Med Bull. 1993 Jul;49(3):506-22
8221019
J Agric Food Chem. 2005 Feb 23;53(4):823-37
15712986
Mini Rev Med Chem. 2002 Jun;2(3):285-93
12370070
Eksp Klin Farmakol. 1997 May-Jun;60(3):41-4
9324397
Biochemistry (Mosc). 1997 Jun;62(6):609-12
9284541