Abstract
Background: Nepeta binaludensis Jamzad is a perennial medicinal plant with various pharmacological effects. However, the neuroprotective effects of Nepeta binaludensis extracts have not yet been investigated. This study aimed to examine the effects of N. binaludensis hydro-alcoholic extract (NBE) on oxidative stress markers and apoptosis-related proteins in PC12 cells exposed to oxygen-glucose deprivation/ reperfusion (OGD/R).
Methods: PC12 cells were pretreated with NBE (at a concentration range of 10-200 μg/ml) before exposure to the OGD condition for 6 h, followed by a 24 h reoxygenation. Cell viability, the production of the reactive oxygen species (ROS), lipid peroxidation (LPO), and the levels of apoptosis-related proteins were evaluated using MTT, fluorimetry, and western blot analysis, respectively.
Results: Survival of the cells preincubated for 6 h with NBE increased to 90.20 ± 15.62% compared with those subjected to OGD/R alone (51.26 ± 7.77 %, p < 0.001). ROS formation was also decreased following incubation with 200 μg/ml of NBE to 125.3 ± 18.38% compared to OGD/R group (356.9 ± 70.48%, p < 0.001). LPO was also suppressed after incubation with NBE to 155.5 ± 21.21% compared to the OGD/R group (260.5 ± 9.727%, p<0.001). NBE restored Bax/Bcl-2 ratio (1.3-fold of control), and cleaved caspase-3 (1.58-fold of control, p < 0.001).
Conclusion: These results suggest that NBE may offer neuroprotection properties against OGD/Rinduced toxicity through modulation of oxidative stress and apoptotic responses.
Keywords: Nepeta binaludensis, oxygen-glucose deprivation/reperfusion (OGD/R), apoptosis, antioxidant, neuroprotection, fluorimetry.
Graphical Abstract
[1]
Majid, A. Neuroprotection in stroke: Past, present, and future. ISRN Neurol., 2014, 2014, 515716.
[http://dx.doi.org/10.1155/2014/515716] [PMID: 24579051]
[http://dx.doi.org/10.1155/2014/515716] [PMID: 24579051]
[2]
Thornton, C.; Leaw, B.; Mallard, C.; Nair, S.; Jinnai, M.; Hagberg, H. Cell death in the developing brain after hypoxia-ischemia. Front. Cell. Neurosci., 2017, 11, 248.
[http://dx.doi.org/10.3389/fncel.2017.00248] [PMID: 28878624]
[http://dx.doi.org/10.3389/fncel.2017.00248] [PMID: 28878624]
[3]
Sims, N.R.; Muyderman, H. Mitochondria, oxidative metabolism and cell death in stroke. Biochim. Biophys. Acta, 2010, 1802(1), 80-91.
[http://dx.doi.org/10.1016/j.bbadis.2009.09.003] [PMID: 19751827]
[http://dx.doi.org/10.1016/j.bbadis.2009.09.003] [PMID: 19751827]
[4]
Liu, Y.; Wu, X.; An, J.; Lv, W.; Geng, Y.; Lou, T.; Zhang, Y. Glaucocalyxin B protects against oxygen-glucose-deprivation/reperfusion-induced neuronal injury in PC-12 cells. J. Cell. Biochem., 2019, 120(4), 6137-6144.
[http://dx.doi.org/10.1002/jcb.27901] [PMID: 30304556]
[http://dx.doi.org/10.1002/jcb.27901] [PMID: 30304556]
[5]
Rajabian, A.; Sadeghnia, H.R.; Hosseini, A.; Mousavi, S.H.; Boroushaki, M.T. 3-Acetyl-11-keto-β-boswellic acid attenuated oxidative glutamate toxicity in neuron-like cell lines by apoptosis inhibition. J. Cell. Biochem., 2020, 121(2), 1778-1789.
[http://dx.doi.org/10.1002/jcb.29413] [PMID: 31642100]
[http://dx.doi.org/10.1002/jcb.29413] [PMID: 31642100]
[6]
Liu, B.; Deng, Q.; Zhang, L.; Zhu, W. Nobiletin alleviates ischemia/reperfusion injury in the kidney by activating PI3K/AKT pathway. Mol. Med. Rep., 2020, 22(6), 4655-4662.
[http://dx.doi.org/10.3892/mmr.2020.11554] [PMID: 33173956]
[http://dx.doi.org/10.3892/mmr.2020.11554] [PMID: 33173956]
[7]
Tayarani-Najaran, Z.; Akaberi, M.; Vatani, M.; Emami, S.A. Evaluation of antioxidant and anti-melanogenic activities of different extracts from aerial parts of Nepeta binaludensis Jamzad in murine melanoma B16F10 cells. Iran. J. Basic Med. Sci., 2016, 19(6), 662-669.
[PMID: 27482348]
[PMID: 27482348]
[8]
Delage, C.; Breard-Mellin, L.; Theresine, C.; Simioneck, S.; Lefranc, B.; Leprince, J.; Bénard, M.; Vaudry, D. The heterogeneity of response of PC12 cell lines from different laboratories to NGF and PACAP questions the reproducibility of studies carried out with tumor cell lines. Neuroendocrinology, 2021.
[http://dx.doi.org/10.1159/000518337]
[http://dx.doi.org/10.1159/000518337]
[9]
Perera, PGT; Bazaka, O; Bazaka, K Pheochromocytoma (PC 12) as a model cell line for membrane permeabilization studies in the presence of electromagnetic fields (EMFs): Recent advances. J. Neurol. Neuromed., 2019, 4(1), 35-40.
[10]
Cao, W.; Feng, S-J.; Kan, M-C. Naringin targets NFKB1 to alleviate oxygen-glucose deprivation/reoxygenation–induced injury in PC12 cells via modulating HIF-1α/AKT/mTOR-signaling pathway. J. Mol. Neurosci., 2021, 71(1), 101-111.
[http://dx.doi.org/10.1007/s12031-020-01630-8] [PMID: 32557145]
[http://dx.doi.org/10.1007/s12031-020-01630-8] [PMID: 32557145]
[11]
Feng, L.; Gao, J.; Liu, Y.; Shi, J.; Gong, Q. Icariside II alleviates oxygen-glucose deprivation and reoxygenation-induced PC12 cell oxidative injury by activating Nrf2/SIRT3 signaling pathway. Biomed. Pharmacother., 2018, 103, 9-17.
[http://dx.doi.org/10.1016/j.biopha.2018.04.005] [PMID: 29635133]
[http://dx.doi.org/10.1016/j.biopha.2018.04.005] [PMID: 29635133]
[12]
Miceli, N.; Taviano, M.F.; Giuffrida, D.; Trovato, A.; Tzakou, O.; Galati, E.M. Anti-inflammatory activity of extract and fractions from Nepeta sibthorpii Bentham. J. Ethnopharmacol., 2005, 97(2), 261-266.
[http://dx.doi.org/10.1016/j.jep.2004.11.024] [PMID: 15707763]
[http://dx.doi.org/10.1016/j.jep.2004.11.024] [PMID: 15707763]
[13]
Jamzad, Z. Nepeta menthoides Boiss. & Buhse and species allied to it in Iran. Iran. J. Bot., 1991, 5(1), 17-27.
[14]
Nadjafi, F.; Koocheki, A.; Rezvani Moghaddam, P.; Rastgoo, M. Ethnopharmacology of Nepeta binaludensis Jamzad a highly threatened medicinal plant of Iran. J. Med. Plants Res., 2009, 8(30), 29-35.
[15]
Amin, M.; Nikoopour, H.; Fazeli, M.R. A survey of antibacterial effects of Oliveria decumbens and Nepeta binaludensis essential oils on Staphylococcus aureus and Escherichia coli in doogh. J. Med. Plants Res., 2019, 18(69), 134-142.
[16]
Mahalleh, A.A.; Sharayei, P.; Azarpazhooh, E. Investigating the characteristics of the Nepeta binaludensis encapsulated extract and its release kinetics in laboratory conditions. Food Bioprocess Technol., 2021, 14(1), 164-176.
[http://dx.doi.org/10.1007/s11947-020-02560-4]
[http://dx.doi.org/10.1007/s11947-020-02560-4]
[17]
Süntar, I.; Nabavi, S.M.; Barreca, D.; Fischer, N.; Efferth, T. Pharmacological and chemical features of Nepeta L. genus: Its importance as a therapeutic agent. Phytother. Res., 2018, 32(2), 185-198.
[http://dx.doi.org/10.1002/ptr.5946] [PMID: 29044858]
[http://dx.doi.org/10.1002/ptr.5946] [PMID: 29044858]
[18]
Khaladgi, M.; Jamzad, M.; Mirahmadpour, P. Total phenolic and flavonoid contents, antioxidant and antimicrobial activity of Nepeta binaludensis Jamzad extracts. Jundishapur J. Nat. Pharm. Prod., 2018, 13(2)
[http://dx.doi.org/10.5812/jjnpp.58500]
[http://dx.doi.org/10.5812/jjnpp.58500]
[19]
Tundis, R.; Nadjafi, F.; Menichini, F. Angiotensin-converting enzyme inhibitory activity and antioxidant properties of Nepeta crassifolia Boiss & Buhse and Nepeta binaludensis Jamzad. Phytother. Res., 2013, 27(4), 572-580.
[http://dx.doi.org/10.1002/ptr.4757] [PMID: 22693035]
[http://dx.doi.org/10.1002/ptr.4757] [PMID: 22693035]
[20]
Rajabian, A.; Sadeghnia, H-R.; Moradzadeh, M.; Hosseini, A. Rheum turkestanicum reduces glutamate toxicity in PC12 and N2a cell lines. Folia Neuropathol., 2018, 56(4), 354-361.
[http://dx.doi.org/10.5114/fn.2018.80869] [PMID: 30786673]
[http://dx.doi.org/10.5114/fn.2018.80869] [PMID: 30786673]
[21]
Inauen, W.; Granger, D.N.; Meininger, C.J.; Schelling, M.E.; Granger, H.J.; Kvietys, P.R. An in vitro model of ischemia/reperfusion-induced microvascular injury. Am. J. Physiol., 1990, 259(1 Pt 1), G134-G139.
[PMID: 2372057]
[PMID: 2372057]
[22]
Li, Y.; Zhang, W.; Chen, C.; Zhang, C.; Duan, J.; Yao, H.; Wei, Q.; Meng, A.; Shi, J. Inotodiol protects PC12 cells against injury induced by oxygen and glucose deprivation/restoration through inhibiting oxidative stress and apoptosis. J. Appl. Biomed., 2018, 16(2), 126-132.
[http://dx.doi.org/10.1016/j.jab.2017.11.004]
[http://dx.doi.org/10.1016/j.jab.2017.11.004]
[23]
Wu, D; Yotnda, P Production and detection of reactive oxygen species (ROS) in cancers. J. Vis. Exp., 2011, 57.
[24]
Buege, J.A.; Aust, S.D. Microsomal lipid peroxidation. Methods Enzymol., 1978, 52, 302-310.
[http://dx.doi.org/10.1016/S0076-6879(78)52032-6] [PMID: 672633]
[http://dx.doi.org/10.1016/S0076-6879(78)52032-6] [PMID: 672633]
[25]
Emami, S.A.; Al-Najjar, H.; Asili, J.; Habibi, M.S.; Sahebkar, A.; Tayarani-Najaran, Z. A comparative study of the cytotoxic and proapoptotic properties of Nepeta binaloudensis. Lett. Drug Des. Discov., 2019, 16(5), 563-569.
[http://dx.doi.org/10.2174/1570180815666180918162805]
[http://dx.doi.org/10.2174/1570180815666180918162805]
[26]
Hou, T.; Zhang, X.; Xu, J.; Jian, C.; Huang, Z.; Ye, T.; Hu, K.; Zheng, M.; Gao, F.; Wang, X.; Cheng, H. Synergistic triggering of superoxide flashes by mitochondrial Ca2+ uniport and basal reactive oxygen species elevation. J. Biol. Chem., 2013, 288(7), 4602-4612.
[http://dx.doi.org/10.1074/jbc.M112.398297] [PMID: 23283965]
[http://dx.doi.org/10.1074/jbc.M112.398297] [PMID: 23283965]
[27]
Singh, A.; Kukreti, R.; Saso, L.; Kukreti, S. Oxidative stress: A key modulator in neurodegenerative diseases. Molecules, 2019, 24(8), 1583.
[http://dx.doi.org/10.3390/molecules24081583] [PMID: 31013638]
[http://dx.doi.org/10.3390/molecules24081583] [PMID: 31013638]
[28]
Liu, L.; Zhao, Z.; Yin, Q.; Zhang, X. TTB protects astrocytes against oxygen-glucose deprivation/reoxygenation-induced injury via activation of Nrf2/HO-1 signaling pathway. Front. Pharmacol., 2019, 10, 792.
[http://dx.doi.org/10.3389/fphar.2019.00792] [PMID: 31379570]
[http://dx.doi.org/10.3389/fphar.2019.00792] [PMID: 31379570]
[29]
Emami, S.A.; Yazdian-Robati, R.; Sadeghi, M.; Baharara, J.; Amini, E.; Salek, F.; Tayarani-Najaran, Z. Inhibitory effects of different fractions of Nepeta satureioides on melanin synthesis through reducing oxidative stress. Res. Pharm. Sci., 2017, 12(2), 160-167.
[http://dx.doi.org/10.4103/1735-5362.202455] [PMID: 28515769]
[http://dx.doi.org/10.4103/1735-5362.202455] [PMID: 28515769]
[30]
Khoshnam, S.E.; Winlow, W.; Farzaneh, M.; Farbood, Y.; Moghaddam, H.F. Pathogenic mechanisms following ischemic stroke. Neurol. Sci., 2017, 38(7), 1167-1186.
[http://dx.doi.org/10.1007/s10072-017-2938-1] [PMID: 28417216]
[http://dx.doi.org/10.1007/s10072-017-2938-1] [PMID: 28417216]
[31]
Ren, Z.; Zhang, R.; Li, Y.; Li, Y.; Yang, Z.; Yang, H. Ferulic acid exerts neuroprotective effects against cerebral ischemia/reperfusion-induced injury via antioxidant and anti-apoptotic mechanisms in vitro and in vivo. Int. J. Mol. Med., 2017, 40(5), 1444-1456.
[http://dx.doi.org/10.3892/ijmm.2017.3127] [PMID: 28901374]
[http://dx.doi.org/10.3892/ijmm.2017.3127] [PMID: 28901374]
[32]
Nadjafi, F; Koocheki, A; Moghaddam, PR; Honermeier, B First experiments on cultivation of Nepeta binaludensis Jamzad, an example of domestication of a highly endangered medicinal plant of Iran. Z ARZNEI- GEWURZPFLA., 2012, 17 64-71. 2012, 17, 64-71.
[33]
Kudo, T.A.; Tominami, K.; Izumi, S.; Hayashi, Y.; Noguchi, T.; Matsuzawa, A.; Hong, G.; Nakai, J. Characterization of PC12 cell subclones with different sensitivities to programmed thermal stimulation. Int. J. Mol. Sci., 2020, 21(21), 8356.
[http://dx.doi.org/10.3390/ijms21218356] [PMID: 33171774]
[http://dx.doi.org/10.3390/ijms21218356] [PMID: 33171774]
[34]
Goldstein, D.S.; Jinsmaa, Y.; Sullivan, P.; Holmes, C.; Kopin, I.J.; Sharabi, Y. 3, 4-Dihydroxyphenylethanol (hydroxytyrosol) mitigates the increase in spontaneous oxidation of dopamine during monoamine oxidase inhibition in PC12 cells. Neurochem. Res., 2016, 41(9), 2173-2178.
[http://dx.doi.org/10.1007/s11064-016-1959-0] [PMID: 27220335]
[http://dx.doi.org/10.1007/s11064-016-1959-0] [PMID: 27220335]
[35]
Li, Z.R.; Yang, L.; Zhen, J.; Zhao, Y.; Lu, Z.N. Nobiletin protects PC12 cells from ERS-induced apoptosis in OGD/R injury via activation of the PI3K/AKT pathway. Exp. Ther. Med., 2018, 16(2), 1470-1476.
[http://dx.doi.org/10.3892/etm.2018.6330] [PMID: 30116396]
[http://dx.doi.org/10.3892/etm.2018.6330] [PMID: 30116396]
[36]
Wiatrak, B.; Kubis-Kubiak, A.; Piwowar, A.; Barg, E. PC12 cell line: Cell types, coating of culture vessels, differentiation and other culture conditions. Cells, 2020, 9(4), 958.
[http://dx.doi.org/10.3390/cells9040958] [PMID: 32295099]
[http://dx.doi.org/10.3390/cells9040958] [PMID: 32295099]
Article Metrics
1