Chemical composition and neuroprotective activity of hemp seed aqueous extract and their chemotaxonomic significance

Abstract

Fructus Cannabis (hemp seed) is important in food and traditional Chinese medicinal applications. Several studies have shown it has antioxidant, antiaging, anti-inflammatory, and neuroprotective properties. Studies have reported its anti-Alzheimer's disease effects. However, its active substances have not been defined, and little is known about the chemical constituents of the aqueous extract. The chemical profile of the aqueous extract of Fructus Cannabis (EFC) was obtained via isolation, structural identification, and qualitative and quantitative analyses. Twenty-seven compounds were identified, including seven nucleosides (1-7), five phenylpropanamides (8-11, and 24), three alkaloids (15, 16, and 26), two cyclic dipeptides (17 and 25), and one pyrimidine (19). Compounds 1, 3-7, 12, 14-19, and 21-27 were not reported previously in the Cannabis genus. Therefore, their chemotaxonomic significance is discussed. Neuroprotective activity screening revealed that EFC and the isolated compounds, particularly 9, 11, and 17, showed significant neuroprotective effects in PC12 ​cells (rat pheochromocytoma cells). The novel object recognition experiment and Nissl staining showed that EFC improved cognitive impairment in APP/PS1 mice and that EFC intervention reduced the number of senile plaques. These findings will contribute to the utility of Fructus Cannabis.

Keywords

Hemp seed / Aqueous extract / Cyclic dipeptides / Neuroprotective effects / Chemotaxonomic significance / APP/PS1 AD model mice

Funding sources

This work was funded by the National Natural Science Foundation of China (grant no. 82073984).

Conflicts of interest

The authors declare no competing financial interests that influence the work reported herein.

Data availability

All data are available upon reasonable requests.

Ethics approval

Animal experiments were approved by the Laboratory Animal Ethical and Welfare Committee of Shandong University Cheeloo College of Medicine (Approval No. 2000-6).

CRediT authorship contribution statement

Siqing Bu: Writing - review & editing, Writing - original draft, Validation, Project administration, Methodology, Investigation. Jianbo Ji: Validation, Resources. Mei Yang: Methodology. Jieyue Sun: Methodology, Formal analysis. Jiaozhen Zhang: Software. Shunmeng Qian: Project administration. Hongxiang Lou: Writing - review & editing, Validation, Funding acquisition. Peihong Fan: Writing - review & editing, Writing - original draft, Project administration, Funding acquisition, Conceptualization.

Declaration of competing interest

The authors declare that they have no known competing financial interests to influence the work reported in this paper.

Acknowledgments

Not applicable.

Appendix A. Supplementary data

Supplementary data to this article can be found online at https://doi.org/10.1016/j.pscia.2024.100051. https://doi.org/10.1016/j.pscia.2024.100051

References

[1] J.G. Zhu, X. Yao, X. Yang, J. Fu, X. Liu, L. Bai, L. Liu, Z. Jiang, (±)-Sativamides A and B, two pairs of racemic nor-lignanamide enantiomers from the Fruits of Cannabis sativa (Hemp Seed), J. Org. Chem. 83 (4) (2018) 1-18. https://10.1021/acs.joc.7b02765.

[2] Q. Wang, Y. Xiong, Processing, nutrition, and functionality of hempseed protein: a Review, Compr. Rev. Food Sci. Food Saf. 18 (4) (2019) 936-952. https://10.1111/1541-4337.12450.

[3] G. Aiello, E. Fasoli, G. Boschin, C. Lammi, C. Zanoni, A. Citterio, A. Arnoldi, Proteomic characterization of hempseed (Cannabis sativa L.), J. Proteom. 147 (2016) 187-196. https://10.1016/j.jprot.2016.05.033.

[4] S. Wang, Q. Luo, P. Fan, Cannabisin F from Hemp (Cannabis sativa) Seed suppresses lipopolysaccharide-induced inflammatory responses in BV2 microglia as SIRT1 modulator, Int. J. Mol. Sci. 20 (3) (2019). https://10.3390/ijms20030507.

[5] X. Yan, J. Tang, C. dos Santos Passos, A. Nurisso, C.A. Simões-Pires, M. Ji, H. Lou, P. Fan, Characterization of lignanamides from Hemp (Cannabis sativa L.) Seed and their antioxidant and acetylcholinesterase inhibitory activities, J. Agric. Food Chem. 63 (49) (2015) 10611-10619. https://10.1021/acs.jafc.5b05282.

[6] N.M. Rodriguez-Martin, R. Toscano, A. Villanueva, J. Pedroche, F. Millan, S. Montserrat-de la Paz, M.C. Millan-Linares, Neuroprotective protein hydrolysates from hemp (Cannabis sativa L.) seeds, Food Funct. 10 (10) (2019) 6732-6739. https://10.1039/c9fo01904a.

[7] S. Moccia, F. Siano, G.L. Russo, M.G. Volpe, F. La Cara, S. Pacifico, S. Piccolella, G. Picariello, Antiproliferative and antioxidant effect of polar hemp extracts (Cannabis sativa L., Fedora cv.)in human colorectal cell lines, Int. J. Food Sci. Nutr. 71 (4) (2020) 410-423. https://10.1080/09637486.2019.1666804.

[8] Z. Lin, L. Chen, Z. Liang, X. Xia, The effect of different extractants from Hemp seed in mice with experimental Alzheimer's disease, Pharm Clin Chin Materia Medica. 32 (6) (2016) 130-134. https://10.13412/j.cnki.zyyl.2016.06.035.

[9] N. Chen, C. Liu, W. Lin, Y. Ding, Z. Bian, L. Huang, H. Huang, K. Yu, S. Chen, Y. Sun, L. Wei, J. Peng, S. Pan, Extract of Fructus Cannabis ameliorates learning and memory impairment induced by D-Galactose in an aging rats model, Evid. Based Compl. Alternat. Med. 2017 (2017), https://doi.org/10.1155/2017/4757520.

[10] Y. Zhou, S. Wang, J. Ji, H. Lou, P. Fan, Hemp (Cannabis sativa L.) seed phenylpropionamides composition and effects on, memory dysfunction and biomarkers of neuroinflammation induced by lipopolysaccharide in mice, ACS Omega 3 (11) (2018) 15988-15995. https://10.1021/acsomega.8b02250.

[11] Z. Rogóź, Effect of combined treatment with mirtazapine and risperidone on the MK-801-induced changes in the object recognition test in mice, Pharmacol. Rep. 65 (5) (2013) 1401-1406. https://10.1016/s1734-1140(13)71499-3.

[12] L. Ruan, H. Tian, R. Jiang, Lipophilic small molecular compounds from human placenta, J. Int. Pharm. Res. 40 (6) (2013) 778-784. https://10.13220/j.cnki.jipr.2013.06.013.

[13] D.L. Domondon, W. He, N. De Kimpe, M. Höfte, J. Poppe, Beta-adenosine, a bioactive compound in grass chaff stimulating mushroom production, Phytochem 65 (2) (2004) 181-187. https://10.1016/j.phytochem.2003.11.004.

[14] Y. Ji, T. Zheng, J. Yin, Chemical constituents from Chinese marine sponge Cinachyrella sp, Chin. J. Mar. Drugs. 37 (2) (2018) 7-12. https://10.13400/j.cnki.cjmd.2018.02.002.

[15] J. Zhou, Y. Jiang, Z. Bi, P. Li, Study on nucleosides from Fritillaria puqiensis, Chin. Pharmaceut. J. (12) (2008) 894-896. https://10.13400/j.cnki.cjmd.2018.02.002.

[16] G. Zhang, H. Ren, Y. Zhang, L. Ma, Y. Yang, S. Wang, Chemical constituents of the starfish Asterias rollestoni Bell, Biochem. Systemat. Ecol. 51 (2013) 203-206. https://10.1016/j.bse.2013.08.031.

[17] X. Chen, B. Zhang, K. Bi, Isolation and identification of chemical components from the buds of Panax notoginseng, J. Shenyang Pharm. Univ. 26 (10) (2009) 775-777. https://10.14066/j.cnki.cn21-1349/r.2009.10.001.

[18] P.M. Moreno, M. Wenska, K.E. Lundin, O. Wrange, R. Strömberg, C.I. Smith, A synthetic snRNA m3G-CAP enhances nuclear delivery of exogenous proteins and nucleic acids, Nucleic Acids Res. 37 (6) (2009) 1925-1935. https://10.1093/nar/gkp048.

[19] W. Zhou, J. Ouyang, G. Li, X. Wang, C. Ding, Y. Suo, H. Wang, Three new alkaloids from Hippophae rhamnoides Linn.subsp. sinensis Rousi, Helv. Chim. Acta 98 (2015) 1287-1291. https://10.1002/chin.201603187.

[20] H. Kuang, B. Yang, Y. Xia, W. Feng, Chemical constituents from the flower of Datura metel, Chin. J. Nat. Med. 8 (6) (2010) 429-431. https://10.3724/SP.J.1009.2010.00429.

[21] Y. Wu, G. Chang, F. Ko, C. Teng, Bioactive constitutents from the stems of Annona montana, Planta Med. 61 (2) (1995) 146-149. https://10.1055/s-2006-958035.

[22] Keith D. Green, Marina Y. Fosso, Sylvie Garneau-Tsodikova, Multifunctional donepezil analogues as cholinesterase and BACE1 inhibitors, Molecules 23 (12) (2018) 22. https://10.3390/molecules23123252.

[23] G. Ma, X. Chen, Z. Hung, T. Wu, X. Xu, X. Zhong, Identification of water-soluble phenolic acids from Clerodendranthus spicatus, Chin. Tradit. Herb. Drugs 48 (13) (2017) 2614-2618. https://10.7501/j.issn.0253-2670.2017.13.005.

[24] Y. Fei, Z. Chen, X. Li, Q. Xu, S, Chemical constituents from seeds of Helianthus annuus, Chin. Tradit. Herb. Drugs 45 (5) (2014) 631-634. https://10.7501/j.issn.0253-2670.2014.05.007.

[25] C. Li, L. Yang, L. Lin, Z. Wang, Z. Li, Chemical constituents from whole plants of Aconitum tanguticum (IV), Chin. J. Exp. Tradit. Med. Formulae 22 (4) (2016) 39-44. https://10.13422/j.cnki.syfjx.2016040039.

[26] G. Zhang, B. Li, H. Cui, L. Chen, Y. Tian, S. Liu, B. Li, M. Li, Z. Xia, X. Chen, Y. Hou, J. Dong, Orychophragines A-C, three biologically active alkaloids from orychophragmus violaceus, Org. Lett. 20 (3) (2018) 656-659. https://10.1021/acs.orglett.7b03801.

[27] Y. Jiang, H. Liang, X. Yuan, J. Yao, R. Qiu, M. Yang, G. Zhang, F. Li, Chemical constituents of jingfang granules based on GC-MS and UPLC-Q exactive MS, Chin. Tradit. Herb. Drugs 53 (6) (2022) 1697-1708. https://10.7501/j.issn.0253-2670.2022.06.012.

[28] J.B. Bremner, W. Sengpracha, I. Southwell, C. Bourke, B.W. Skelton, A.H. White, A revised structure for the alkaloid, tribulusterine, from Tribulus terrestris L, Aust. J. Chem. 57 (3) (2004) 273-276. https://10.1071/ch03230.

[29] Vanessa L. Campo, Maristela B. Martins, Carlos H.T.P. da Silva, Ivone Carvalho, Novel and facile solution-phase synthesis of 2,5-diketopiperazines and Oglycosylated analogs, Tetrahedron 65 (2009) 5343-5349. https://10.1016/j.tet.2009.04.069.

[30] EMaria Harizani, Eleni Katsini, Panagiota Georgantea, Vassilios Roussis, Efstathia Ioannou, New chlorinated 2,5-diketopiperazines from marine-derived bacteria isolated from sediments of the eastern mediterranean sea, Molecules 25 (7) (2020). https://10.3390/molecules25071509.

[31] J. Gao, J. Sheng, X. Yang, J. Liu, The constituents of Russula ochroleuca basidiomycetes, Acta Bot. Yunnanica (2001) 385-393. https://10.3969/j.issn.2095-0845.2001.03.019.

[32] K. Guo, X. Wang, J. Guo, J. Li, J. Lu, Chemical constituents from leaves of Elaeagnus mollis, Chin. Tradit. Herb. Drugs 48 (2) (2017) 236-240. https://10.7501/j.issn.0253-2670.2017.02.003.

[33] Z. Ding, L. Hu, J. Liu, The chemical constituents of basidiomycetes Boletopsis grisea, Acta Bot. Yunnanica 24 (5) (2002) 667-670. https://10.3969/j.issn.2095-0845.2002.05.015.

[34] K. Ioanna, Sideri Errika Voutyritsa Christoforos G. Kokotos, Green photoorganocatalytic synthesis of phenols from arylboronic acids, Synlett 29 (10) (2018) 1324-1328. https://10.1055/s-0036-1591837.

[35] F. Yang, R. Chen, L. Feng, H. Li, H. Zhang, J. Liang, Chemical constituents from the aerial part of Peganum nigellastrum, Chin. J. Nat. Med. 8 (3) (2010) 199-201. https://10.3724/sp.J.1009.2010.00199.

[36] L.H.N. de Sousa, R.D. de Araújo, D. Sousa-Fontoura, F.G. Menezes, R.M. Araújo, Metabolities from marine sponges of the genus Callyspongia: occurrence, biological activity, and NMR data, Mar. Drugs 19 (12) (2021). https://10.3390/md19120663.

[37] Y. Zhao, X. Wu, N. Yu, A novel analgesic pyrazine derivative from the leaves of Croton tiglium L, J. Asian Nat. Prod. Res. 9 (5) (2007) 437-441. https://10.1080/10286020500384781.

[38] F. Kong, P. Wang, J. Wei, Y. Wang, W. Wang, K. Hong, W. Zhu, Alkaloids from the mangrove-derived actinomycete Jishengella endophytica 161111, Mar. Drugs 12 (2014) 477-490. https://10.3390/md12010477.

[39] M.S. Alavi, S. Fanoudi, A. Hosseini, M. Jalili-Nik, A. Bagheri, H.R. Sadeghnia, Everolimus attenuates glutamate-induced PC12 cells death, Int. J. Neurosci. 133 (4) (2023) 457-466. https://10.1080/00207454.2021.1929210.

[40] L. Ding, D. Li, L. Yang, F. Qiu, Nitrogenous compounds from Sargassum pallidum, Chin. Tradit. Herb. Drugs 48 (9) (2017) 1735-1739. https://10.7501/j.issn.0253-2670.2017.09.007.

[41] P. Xu, W. Du, B. Liu, X. Xu, X. Lai, B. Li, Chemical constituents from shoots of Phyllostachys edulis (I), Chin. Tradit. Herb. Drugs 46 (3) (2015) 334-338. https://10.7501/j.issn.0253-2670.2015.03.005.

[42] Q. Li, H. Liang, B. Wang, Y. Zhao, Chemical constituents of Momordica charantia L, Acta Pharm. Sin. 44 (9) (2009) 1014-1018. https://10.16438/j.0513-4870.2009.09.006.

[43] W. Guo, S. Guo, Chemical composition study of endophytic fungus Epulorhiza sp. with potential anti HIV activity, Chin. Tradit. Herb. Drugs 41 (11) (2010) 1773-1775. https://CNKI:SUN:ZCYO.0.2010-11-006.

[44] H. Wang, Y. Hu, W. Ye, G. Liu, Aqueous constituents from Stellaria media (L.) cyr, J. China Pharm. Univ. 36 (6) (2005) 523-525, https://doi.org/10.3321/j.issn:1000-5048.2005.06.009.

[45] T. Xu, X. Zhou, K. Wen, X. Yang, S. Xu, Y. Liu, New N-acyl taurine from the sea urchin Glyptocidaris crenularis, Biotechnol. Biochem. 74 (5) (2010) 1089-1091. https://10.1271/bbb.90848.