Cell Stress and Chaperones

, Volume 23, Issue 4, pp 763–772 | Cite as

HO-1 dependent antioxidant effects of ethyl acetate fraction from Physalis alkekengi fruit ameliorates scopolamine-induced cognitive impairments

  • Md. Moniruzzaman
  • Young-Won Chin
  • Jungsook Cho
Original Paper


Physalis alkekengi var. francheti is an indigenous herb well known for its anti-inflammatory, sedative, antipyretic, and expectorant properties. However, the information regarding the impacts of P. alkekengi fruits (PAF) in modulation of oxidative stress and learning memory are still unknown. This study therefore evaluated the antioxidant properties of ethyl acetate (EA) fraction of PAF and its impacts on learning and memory. The antioxidant activities of PAF were evaluated in LPS-induced BV2 microglial cells. The potent EA fraction then investigated and confirmed for its involvement of HO-1 pathway using hemin (HO-1 inducer) and ZnPP (HO-1 inhibitor) through Western blotting, DCFH-DA, and/or Griess assay. The involvements of PI3K/Akt, MEK, and p38 MAPK also investigated. Furthermore, we applied EA fraction to the animals at 100 and 200 mg/kg doses to check if the extract could improve scopolamine-induced memory deficits in passive avoidance and elevated plus maze tests. Our results demonstrated that the fractions from PAF significantly inhibited the generation of intracellular reactive oxygen species (ROS) induced by LPS in concentration-dependent manners. In comparison to other fractions, the EA fraction exhibited potent effect in suppressing intracellular ROS generation. Besides, EA fraction also induced the expression of HO-1 in time- and concentration-dependent manners. ZnPP significantly reversed the suppressive effect of EA fraction on LPS-induced ROS generation and NO production, which confirm the involvement of HO-1 signaling in EA-fraction-mediated antioxidant activities. Consistently, blocking of PI3K/Akt, MEK, and p38 MAPK pathways by PAF-EA suppressed the production of intracellular ROS, indicating their potential participation. In addition, one of the major constituents of EA fraction, luteolin-7-O-β-D-glucoside, also demonstrated HO-1-dependent antioxidant effects in BV2 cells. Further, the EA fraction significantly (p < 0.05) improves scopolamine-induced memory deficits in mice. Taken together, our findings highlight the antioxidant effects of EA fraction of PAF which may be beneficial in treatment of different neurodegenerative diseases associated with free radicals.


Physalis alkekengi Solanaceae Luteolin-7-O-β-D-glucoside Antioxidant HO-1 (Hsp-32) Scopolamine Memory deficit 



Heme oxygenase 1


Alzheimer’s disease


Central nervous system




Reactive oxygen species


Hydrogen per oxide


Physalis alkekengi fruit


Mitogen-activated protein kinases


2′,7′-dichloro dihydro fluorescein diacetate


3-(4,5-dimethyldiazol-2-yl)-2,5-diphenyl tetrazolium bromide


Dulbecco’s modified eagle’s medium


Fetal bovine serum


Dimethyl sulfoxide


Nitric oxide


Elevated plus maze


Zinc protoporphyrine



Md. Moniruzzaman was supported by SRD-II scholarship from Dongguk University. Jungsook Cho was funded by the GRRC Program of Gyeonggi province [GRRC DONGGUK2016-B01], Republic of Korea.

Compliance with ethical standards

All experimental protocols used in this study were performed following “Guide for the Care and Use of Laboratory Animals, Institute of Laboratory Animal Resources, Commission on Life Sciences, National Research Council, USA” (National Academy Press: Washington D.C., 1996) and approved by the Institutional Animal Ethical Committee of Dongguk University (Approval Number: IACUC-2013-0005).

Competing interests

Author does not have any conflict of interests associated with this publication.

Supplementary material

12192_2018_887_MOESM1_ESM.docx (152 kb)
ESM 1 (DOCX 152 kb)


  1. Asilbekova DT, Ul’chenko NT, Glushenkova AI (2016) Lipids from Physalis alkekengi. Chem Nat Comp 52(1):96–97. CrossRefGoogle Scholar
  2. Balmus IM, Ciobica A, Antioch I, Dobrin R, Timofte D (2016) Oxidative stress implications in the affective disorders: main biomarkers, animal models relevance, genetic perspectives, and antioxidant approaches. Oxidative Med Cell Longev 2016:1–25. CrossRefGoogle Scholar
  3. Barron KD (1995) The microglial cell. A historical review. J Neurol Sci 134(Suppl):57–68CrossRefPubMedGoogle Scholar
  4. Block ML, Hong JS (2005) Microglia and inflammation-mediated neurodegeneration: multiple triggers with a common mechanism. Prog Neurobiol 76(2):77–98. CrossRefPubMedGoogle Scholar
  5. Chien CC, Shen SC, Yang LY, Chen YC (2012) Prostaglandins as negative regulators against lipopolysaccharide, lipoteichoic acid, and peptidoglycan-induced inducible nitric oxide synthase/nitric oxide production through reactive oxygen species-dependent heme oxygenase 1 expression in macrophages. Shock (Augusta, Ga) 38(5):549–558. CrossRefGoogle Scholar
  6. Deb D, Bairy KL, Nayak V, Rao M (2015) Comparative effect of Lisinopril and Fosinopril in mitigating learning and memory deficit in scopolamine-induced amnesic rats. Adv Pharmacol Sci 2015:521718–521711. PubMedPubMedCentralCrossRefGoogle Scholar
  7. Fan Y, Hu J, Li J, Yang Z, Xin X, Wang J, Ding J, Geng M (2005) Effect of acidic oligosaccharide sugar chain on scopolamine-induced memory impairment in rats and its related mechanisms. Neurosci Lett 374(3):222–226. CrossRefPubMedGoogle Scholar
  8. Gamal-Eldeen AM, Kawashty SA, Ibrahim LF, Shabana MM, El-Negoumy SI (2004) Evaluation of antioxidant, anti-inflammatory, and antinociceptive properties of aerial parts of Vicia Sativa and its flavonoids. J Nat Rem 4(1):81–96Google Scholar
  9. Hanrott K, Gudmunsen L, O'Neill MJ, Wonnacott S (2006) 6-hydroxydopamine-induced apoptosis is mediated via extracellular auto-oxidation and caspase 3-dependent activation of protein kinase Cdelta. J Biol Chem 281(9):5373–5382. CrossRefPubMedGoogle Scholar
  10. Herrera-Morales W, Mar I, Serrano B, Bermudez-Rattoni F (2007) Activation of hippocampal postsynaptic muscarinic receptors is involved in long-term spatial memory formation. Eur J Neurosci 25(5):1581–1588. CrossRefPubMedGoogle Scholar
  11. Ishii T, Itoh K, Takahashi S, Sato H, Yanagawa T, Katoh Y, Bannai S, Yamamoto M (2000) Transcription factor Nrf2 coordinately regulates a group of oxidative stress-inducible genes in macrophages. J Biol Chem 275(21):16023–16029CrossRefPubMedGoogle Scholar
  12. Jung WY, Kim H, Park HJ, Jeon SJ, Park HJ, Choi HJ, Kim NJ, Jang DS, Kim DH, Ryu JH (2016) The ethanolic extract of the Eclipta Prostrata L. ameliorates the cognitive impairment in mice induced by scopolamine. J Ethnopharmacol 190:165–173. CrossRefPubMedGoogle Scholar
  13. Kim S, Lee Y, Cho J (2014) Korean red ginseng extract exhibits neuroprotective effects through inhibition of apoptotic cell death. Biol Pharm Bull 37(6):938–946CrossRefPubMedGoogle Scholar
  14. Kim S, Chin YW, Cho J (2017) Protection of cultured cortical neurons by Luteolin against oxidative damage through inhibition of apoptosis and induction of Heme Oxygenase-1. Biol Pharm Bull 40(3):256–265. CrossRefPubMedGoogle Scholar
  15. Kranjc E, Albreht A, Vovk I, Makuc D, Plavec J (2016) Non-targeted chromatographic analyses of cuticular wax flavonoids from Physalis Alkekengi L. J Chromatogr A 1437:95–106. CrossRefPubMedGoogle Scholar
  16. Li A-L, Chen B-J, Li G-H, Zhou M-X, Li Y-R, Ren D-M, Lou H-X, Wang X-N, Shen T (2018) Physalis Alkekengi L. Var. Franchetii (mast.) Makino: an ethnomedical, phytochemical and pharmacological review. J Ethnopharmacol 210(Supplement C):260–274. CrossRefPubMedGoogle Scholar
  17. Lin W, Wu RT, Wu T, Khor TO, Wang H, Kong AN (2008) Sulforaphane suppressed LPS-induced inflammation in mouse peritoneal macrophages through Nrf2 dependent pathway. Biochem Pharmacol 76(8):967–973. CrossRefPubMedPubMedCentralGoogle Scholar
  18. Moniruzzaman M, Bose S, Kim YM, Chin YW, Cho J (2016) The ethyl acetate fraction from Physalis Alkekengi inhibits LPS-induced pro-inflammatory mediators in BV2 cells and inflammatory pain in mice. J Ethnopharmacol 181:26–36. CrossRefPubMedGoogle Scholar
  19. Morrison JH, Hof PR (1997) Life and death of neurons in the aging brain. Science (New York, NY) 278(5337):412–419CrossRefGoogle Scholar
  20. Nie K, Yu JC, Fu Y, Cheng HY, Chen FY, Qu Y, Han JX (2009) Age-related decrease in constructive activation of Akt/PKB in SAMP10 hippocampus. Biochem Biophys Res Commun 378(1):103–107. CrossRefPubMedGoogle Scholar
  21. Otterbein LE, Bach FH, Alam J, Soares M, Tao Lu H, Wysk M, Davis RJ, Flavell RA, Choi AM (2000) Carbon monoxide has anti-inflammatory effects involving the mitogen-activated protein kinase pathway. Nat Med 6(4):422–428. CrossRefPubMedGoogle Scholar
  22. Qiu L, Zhao F, Jiang ZH, Chen LX, Zhao Q, Liu HX, Yao XS, Qiu F (2008) Steroids and flavonoids from Physalis Alkekengi Var. Franchetii and their inhibitory effects on nitric oxide production. J Nat Prod 71(4):642–646. CrossRefPubMedGoogle Scholar
  23. Reiter RJ (1995) Oxidative processes and antioxidative defense mechanisms in the aging brain. FASEB J 9(7):526–533CrossRefPubMedGoogle Scholar
  24. Ryter SW, Otterbein LE, Morse D, Choi AMK (2002) Heme oxygenase/carbon monoxide signaling pathways: regulation and functional significance. Mol Cell Biochem 234(1):249–263. CrossRefPubMedGoogle Scholar
  25. Ryter SW, Alam J, Choi AM (2006) Heme oxygenase-1/carbon monoxide: from basic science to therapeutic applications. Physiol Rev 86(2):583–650. CrossRefPubMedGoogle Scholar
  26. Sharma AC, Kulkarni SK (1992) Evaluation of learning and memory mechanisms employing elevated plus-maze in rats and mice. Prog Neuro-Psychopharmacol Biol Psychiatry 16(1):117–125CrossRefGoogle Scholar
  27. Song YS, Park CM (2014) Luteolin and luteolin-7-O-glucoside strengthen antioxidative potential through the modulation of Nrf2/MAPK mediated HO-1 signaling cascade in RAW 264.7 cells. Food Chem Toxicol 65:70–75. CrossRefPubMedGoogle Scholar
  28. Syapin PJ (2008) Regulation of haeme oxygenase-1 for treatment of neuroinflammation and brain disorders. Br J Pharmacol 155(5):623–640. CrossRefPubMedPubMedCentralGoogle Scholar
  29. Takagi T, Naito Y, Uchiyama K, Yoshikawa T (2010) The role of heme oxygenase and carbon monoxide in inflammatory bowel disease. Redox Rep 15(5):193–201. CrossRefPubMedGoogle Scholar
  30. Terry AV Jr, Callahan PM, Hall B, Webster SJ (2011) Alzheimer's disease and age-related memory decline (preclinical). Pharmacol Biochem Behav 99(2):190–210. CrossRefPubMedPubMedCentralGoogle Scholar
  31. Wu ML, Ho YC, Lin CY, Yet SF (2011) Heme oxygenase-1 in inflammation and cardiovascular disease. Am J Cardiovasc Dis 1(2):150–158PubMedPubMedCentralGoogle Scholar
  32. Yang H, Han S, Zhao D, Wang G (2014) Adjuvant effect of polysaccharide from fruits of Physalis Alkekengi L. in DNA vaccine against systemic candidiasis. Carbohydr Polym 109:77–84. CrossRefPubMedGoogle Scholar

Copyright information

© Cell Stress Society International 2018

Authors and Affiliations

  1. 1.College of PharmacyDongguk University-SeoulGoyangRepublic of Korea
  2. 2.Mater Research Institute - The University of Queensland, Translational Research InstituteBrisbaneAustralia
  3. 3.College of Pharmacy and BK-Plus TeamDongguk University-SeoulGoyangRepublic of Korea

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