Evaluation of antioxidant and cytoprotective activities of Artemisia ciniformis extracts on PC12 cells

Document Type : Original Article


1 Pharmaceutical Sciences Research Center, School of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, Iran

2 Students Research Committee, Kermanshah University of Medical Sciences, Kermanshah, Iran


Objective(s): In the current study antioxidant capacities of five different extracts of Artemisia ciniformis aerial parts were evaluated by cell-free methods. Then seven fractions of the potent extract were selected and their antioxidant capacity was assayed by cell free and cell based methods.
Materials andMethods: Antioxidant ability was measured using the: 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging test, β-carotene bleaching (BCB) method and ferrous ion chelating (FIC) assay. Total phenolic contents (TPC) of all the samples also were determined. The cytoprotective effect of fractions was evaluated by measuring the viability of cells after exposure to doxorubicin (DOX). The mechanism of action was studied by investigating caspase-3, mitochondrial membrane potential (MMP), the level of super-oxide dismutase (SOD) and intracellular reactive oxygen species (ROS).
Results: Hydroethanolic extract exhibited a notably higher antioxidant activity and phenolic content. Among the fractions (A to G) of hydroethanolic extract, the highest antioxidant capacity was observed in the Fraction E. Moreover, 24 hr pretreatment of PC12 cells with fractions B, C and D decreased DOX-induced cytotoxicity. In addition, pre-treatment of cells with fraction B resulted in significant decrease in generation of the reactive oxygen species (ROS) and increase in the activity of SOD. We were able to demonstrate remarkable reduction in the activity of caspase-3 and increase in MMP in PC12 cells following pretreatment with fraction B.
Conclusion: Our observations indicated that the fraction B of A. ciniformis hydroetanolic extract possessed protective effect on oxidative stress and apoptosis induced by DOX in PC12 cells.


1.Halliwell B. Free radicals and antioxidants:a  personal view. Nut Rev 1994; 52:253-265.
2.Sies H. Oxidative stress: oxidants and antioxidants. Exp Physiol 1997; 82:291-295.
3.Clark SF. The biochemistry of antioxidants revisited. Nut Clin Pract 2002; 17:5-17.
4. Farzaeia MH, Rahimia R, Attarb F, Siavoshic M, Sanieec P, Hajimahmoodid M, et al. Chemical composition, antioxidant and antimicrobial activity of essential oil and extracts of tragopogon graminifolius, a medicinal herb from Iran. Nat Prod Commun 2014; 9:121-124.
5. Ferguson LR. Role of plant polyphenols in genomic stability. Mutat Res 2001; 475:89-111.
6. Ghaffari H, Ghassam BJ, Chandra  Nayaka S, Ramachandra Kini K, Prakash HS. Antioxidant and neuroprotective activities of Hyptissuaveolens., L. Poit. against oxidative stress-induced neurotoxicity. Cell Mul Neurobiol 2014; 34:323-331.
7. Collins AR. Antioxidant intervention as a route to cancer prevention. Eur J Can 2005; 41:1923-1930.
8. Kundu JK, Chun KS. The Promise of dried fruits in cancer chemoprevention. Asian Pac J Cancer Prev 2014; 15:3343-3352.
9. Ignat I, Volf I, Popa VI. A critical review of methods for characterization of polyphenolic compounds in fruits and vegetables. Food Chem 2011; 126:1821-1835.
10. Rice-Evans C, Miller N, Paganga G. Antioxidant properties of phenolic compounds. Trends Plant Sci 1997; 2:152-159.
11. Velioglu Y, Mazza G, Gao L, Dave OB. Antioxidant activity and total phenolics in selected fruits, vegetables, and grain products. J Agric food Chem 1998; 46:4113-4117.
12. Kähkönen MP, Hopia AI, Vuorela HJ. Antioxidant activity of plant extracts containing phenolic compounds. J Agric Food Chem 1999; 47: 3954-3962.
13. Drolet G, Dumbroff E, Legge R, Thompson JE. Radical scavenging properties of polyamines. Phytochemistry 1986; 25:367-371.
14. Hussain SS, Ali M, Ahmad M, Siddique GH. Polyamines: natural and engineered abiotic and biotic stress tolerance in plants. Biotechnol Adv 2011; 29:300-311.
15. Topçu G, Ertas A, Kolak U. Antioxidant activity tests on novel triterpenoids from Salvia macrochlamys. ARKIVOC 2007; 7:195-208.
16. Xia Q, Zhang H, Sun X, Xia Q Zhao H, Wu L, et al. A comprehensive review of the structure elucidation and biological activity of triterpenoids from Ganoderma spp. Molecules 2014; 19:17478-17535.
17. Wang J, Zhang Q, Zhang Z, Li Z. Antioxidant activity of sulfated polysaccharide fractions extracted from Laminaria japonica. Int J Biol Macromol 2008; 42:127-132.
18. Chang SC, Hsu BY, Chen BH. Structural characterization of polysaccharides from Zizyphusjujuba and evaluation of antioxidant activity. Int J Biol Macromol 2010; 47:445-453.
19. Liang Z, Guo YT, Yi YJ. Ganodermalucidum polysaccharides target a Fas/caspase dependent pathway to induce apoptosis in human colon cancer cells. Asian Pac J Cancer Prev 2014; 15:3981-3986.
20. Bora KS, Sharma A. The Genus artemisia: a comprehensive review. Pharm Biol 2011; 49:101-109.
21. Mozaffarian V. A dictionary of Iranian plant names: Latin, English, Persian. Farhang Mo'aser; 1996.
22. Ghahreman A, Attar F. Biodiversity of plant species in Iran, Central Herbarium of Tehran University, Faculty of Science.1999.
23. Rustaiyan A, Masoudi S, Kazemi M. Volatile oils constituents from different parts of Artemisia ciniformis Krasch. et M. Pop. exPoljak and Artemisia incana .,L. Druce. from Iran. J Essen Oil Res 2007; 19: 548-551.
24. Firouzni A, Vahedi H, Sabbaghi F. Composition of the essential oil of Artemisia ciniformis, A. kopetdaghensis, and A. khorasanica in Iran. Chem Nat Comp 2008; 44:804-806.
25. Emami A, Zamani Tahgizadeh Rabe SH, Ahi A, Mahmoudi M. Study on toxic effects of Artemisisa spp. fractions from Iran on human cancer cell lines. J Zan Univ Med Sci 2010; 18:58-67.
26. Taghizadeh Rabe SZ, Mahmoudi M, Ahi A. Antiproliferative effects of extracts from Iranian Artemisia species on cancer cell lines. Pharm Biol 2011; 49:962-969.
27. Tayarani-Najaran Z, Hajian Z, Mojarrab M Emami A. Cytotoxic and apoptotic effects of extracts of Artemisia ciniformis Krasch.& Popov ex Poljakov on K562 and HL-60 cell lines. Asian Pac J Cancer Prev 2014; 15:7055-7059.
28. Emami SA, Zamanai Taghizadeh Rabe S, Ahi A, Mahmoudi M. Inhibitory Activity of Eleven Artemisia Species from Iran against Leishmania Major Parasites. Iran J Bas Med Sci 2012; 15:807-811.
29. Mojarrab M, Naderi R Heshmati Afshar F. Screening of different etracts from artemisia species for their potential antimalarial activity. Iran J Pharm Res 2015; 14:605-608.
30. Hosseinzadeh L, Behravan J, Mosaffa F, Bahrami G, Bahrami AR, Karimi GH. Curcumin potentiates doxorubicin-induced apoptosis in H9c2 cardiac muscle cells through generation of reactive oxygen species. Food Chem Toxicol 2011; 49:1102-1109.
31. Lopes AM, Meisel M, Dirnagl D, Carvalho FD, Bastos MDL. “Doxorubicin induces biphasic neurotoxicity to rat cortical neurons. Neuro Toxico 2008; 29: 286–293.
32. Park ES, Kim SD, Lee MH,  LeeH S, Lee IS,  Sung JK, et al. Protective effects of N-acetylcysteine and selenium against doxorubicin toxicity in rats. J Vet Sci 2003; 4:129–136.
33. Shokoohinia Y, Hosseinzadeh L, Moieni-Arya M, Mostafaie A, Mohammadi-Motlagh HR. Osthole attenuates doxorubicin-induced apoptosis in PC12 cells through inhibition of mitochondrial dysfunction and ROS production. Biomed Res Int 2014; 2014:156848.
34. Mojarrab M, Jamshidi M, Ahmadi F, Hosseinzadeh L. Extracts of Artemisia ciniformis Protect cytotoxicity induced by hydrogen peroxide in H9c2 cardiac muscle Cells through the Inhibition of Reactive Oxygen Species. Adv Pharmacol Sci 2013; 2013:141683.
35. Mendonc LM¸ Venancio VP,  Bianchi ML, Antunes LM.  Machado CS. Co enzyme Q10 protects PC12 cells from cisplatin-induced DNA damage and neurotoxicity. Neurotoxicology 2013; 36:10–16.
36. Hatano T, Edamatsu R, Hiramatsu M, Moti A, Fujita Y, Yasushara T. Effects of the interaction of tannins with co-existing substances. VI: Effects of tannins and related polyphenols on superoxide anion radical, and on 1, 1-diphenyl-2-picrylhydrazyl radical. Chem Pharm Bull 1989; 37:2016-2021.
37. Hatami T, Emami SA, Miraghaee SS, Mojarrab M. Total phenolic contents and antioxidant activities of different extracts and fractions from the aerial parts of Artemisia biennis Willd. Iran J Pharm Res 2014; 13:551-558.
38. Shokoohinia Y, Rashidi M, Hosseinzadeh L, Jelodarian J. Quercetin-3-O- b-D-glucopyranoside, a dietary flavonoid, protects PC12 cells from H2O2-induced cytotoxicity through inhibition of reactive oxygen species. Food Chem 2015; 167:162–167.
39. Malstorm B, Andreasson l, Reinhammer B. The enzymes. Boyer P. editor XIIB. New York: Academic Press; 1975.p. 533.
40. Sankari SL, Masthan K, Babu NA, Kasturiv M. Apoptosis in cancer–an update. Asian Pac J Cancer Prev 2015; 13:4873-4878.
41. Liu X, Wang J, Lu C, Liu Y. The role of lysosomes in BDE 47-mediated activation of mitochondrial apoptotic pathway in HepG2 cells. Chemosphere 2015; 124:10-21.
42. Hosseinzadeh L, Khorand A, Aliabadi A.  Discovery of 2- Phenyl –N-., 5-(trifluoromethyl)– 1,3,4-thiadiazol-2-yl) acetamid derivatives as apoptosis inducer via caspase pathway whit potential anticancer activity. Arch Pharm Chem Life Sci 2013; 346:812–818.
43.Taveira M, Sousa C, Valentão P, Fereress F, Teixeria JP, Andreid PB. Neuroprotective effect of steroidal alkaloids on glutamate-induced toxicity by preserving mitochondrial membrane potential and reducing oxidative stress. J Asian Nat Prod Res 2013; 16:192-199.
44. Wu WM, Zhu YY, Li HR, Yu HZ, Zhang P, Pi HF. Two new alkaloids from the bulbs of Lycorissprengeri. Steroid Biochem Mol Biol 2014; 140:106-115.
45. Kanhere R, Anjana A, Anbu J. Neuroprotective and antioxidant potential of terpenoid fraction from Hygrophila auriculata against transient global cerebral ischemia in rats. Pharm Biol 2013; 51:181-189.
46. Zhang ZC, Su G, Li J, Xcie XD.  Two new neuroprotective phenolic compounds from Gastro dia- elata. J  Asian Nat Prod Res 2013; 15:619-623.
47. Kelsey NA, Wilkins HM, Linseman DA. Nutraceutical antioxidants as novel neuroprotective agents. Molecules 2010; 15:7792-7814.
48. Kim YC. Neuroprotective phenolics in medicinal plants. Arch Pharmacol Res 2010; 33:1611-1632.
49. Bors W, Heller W, Michel C. Flavonoids and polyphenols: Chem Biol. Antioxidants Health Dis Series 1996; 409-468.
50. Zhang W, Wang J, Jin W, Zhang Q. The antioxidant activities and neuroprotective effect of polysaccharides from the starfish Asteriasrollestoni. Carbohyd Polym 2013; 5:95:9-15.
51. Shi J, Li CJ, Yang JZ, Yuan YH, Chen MH, Zhang DM. Coumarin glycosides and iridoidGlucosides with neuroprotective effects from hydrangea paniculata. Planta Med 2012; 78:1844-1850.
52. Sheng GQ, Zhang JR, Pu XP. Protective effect of verbascoside on 1-methyl-4-phenylpyridinium ion-induced neurotoxicity in PC12 cells. Eur J Pharmacol 2002; 451:119-124.
53. Sarvestani NN, Khodagholi F, Ansari N, Farimini MN. Involvement of p-CREB and phase II detoxifying enzyme system in neuroprotection mediated by the flavonoid calycopterin isolated from Dracocephalumkotschyi. Phytomedicine 2013; 20:939-946.
54. Jin W, Zhang W, Wang J, Yao J, Xie E, Liu D, et al.  A study of neuroprotective and antioxidant activities of heteropolysaccharides from six Sargassum species. Int J Biol Macromol 2014; 67:336-342.
55. Hu W, Wang G, Li P, Wang Y, Si CL, He J, et al. Antioxidant properties and neuroprotective effects of isocampneoside II on hydrogen peroxide-induced oxidative injury in PC12 cells. Food Chem Toxicol 2013; 59:145-152.
56. Jin W, Zhang W, Wang J, Zhang Q. The neuroprotective activities and antioxidant activities of the polysaccharides fromSaccharina japonica. Int J Biol Macromol 2013; 58:240-244.
57. Ji X, Rivers L, Zielinski, MacDugal E, Jancy S, Zhang S. Quantitative analysis of phenolic components and glycoalkaloids from 20 potato clones and in vitro evaluation of antioxidant, cholesterol uptake, and neuroprotective activities. Food Chem 2011; 133:1177-1188.
58. Naik GH, Priyadarsini KI, Satav JG, Banavalikar MM, Sohoni DP, Biyani MK, et al. Comparative antioxidant activity of individual herbal components used in Ayurvedic medicine. Phytochemistry 2003; 63:97–104.
59. Djidel S, khennouf S. Radical scavenging, reducing power, lipid peroxidation inhibition and chelating properties of extracts froma artemisia campestris L. Aerial Parts. Ann Res Rev Biol 2014; 4:1691-1702.
60. Zheng W, Wang SY. Antioxidant activity and phenolic compounds in selected herbs. J Agric Food Chem 2001; 49:5165-5170.