Active compounds and anti-inflammatory activity of the methanolic extracts of the leaves and callus from Tilia americana var. mexicana propagated plants

  • Karen Flores-Sánchez
  • Francisco Cruz-Sosa
  • Alejandro Zamilpa-Alvarez
  • Pilar Nicasio-TorresEmail author
Original Article


Tilia americana var. mexicana is used in Mexican traditional medicine to treat anxiety and inflammatory processes. Several glycosides derived from quercetin and kaempferol, including tiliroside, isoquercetin, and quercetin-3-β-d-glucoside, were reported as the main anxiolytic compounds in this species; to our knowledge, compounds with anti-inflammatory effects have not been previously described. In this study, whole plants were obtained from rooted cuttings with indole-3-butyric acid (IBA) under greenhouse conditions. Multiple shoots and callus cultures were established from apical and axillary buds from T. americana var. mexicana cuttings. The apical buds (75%) were the best explant for shoot induction (2–3 shoots per explant) on Murashige and Skoog (MS) medium supplemented with 2.0 mg L−1 of 6-benzyl aminopurine plus 0.25 mg L−1 α-naphthaleneacetic acid. Callogenesis occurred in both types of buds in the treatments constituted by thidiazuron with 0.1 mg L−1 IBA. High-performance liquid chromatography analysis of leaves and callus methanolic extracts allowed the identification of quercetin-3-β-d-glucoside and tiliroside anxiolytic compounds, and of the scopoletin anti-inflammatory compound. The methanolic leaf and callus extracts showed anti-inflammatory activities in a 12-O-tetradecanoylphorbol-13-acetate-induced ear edema model with median effective doses (ED50) of 0.38 and 1.73 mg per ear for the leaf and callus extracts, respectively.


Anti-inflammatories Anxiolytics Propagation Quercetin-3-β-d-glucoside Scopoletin Tiliroside 



6-Benzyl amino-9-(2-tetrahydropyranyl)-9H-purine


Indole-3-butyric acid


6-Benzyl aminopurine




Murashige and Skoog


α-Naphthaleneacetic acid








2,4-Dichlorophenoxyacetic acid



This work was supported by Basic Grant 593703 from the Consejo Nacional de Ciencia y Tecnología, México (CONACyT-México) for the Doctoral studies of Karen Flores-Sánchez at the Biotechnology Doctoral Program of UAM-Iztapalapa; and by Complementary Grant 99182548 from the IMSS.

Author contributions

As a Ph.D. student, KF-S participated in all of the experimental work, in the collection, analysis, and interpretation of data, and in the writing of the manuscript. FC-S supervised the establishment of the factorial design experiments, provided the scopoletin, quercetin-3-β-d-glucoside, and tiliroside standards, and was the Thesis Co-Director of KF-S. A-Z participated in the extraction and in establishment of analytical methods for the quantification of compounds. PN-T performed the supervision of the establishment of in-vitro cultures and the evaluation of anti-inflammatory activity, and also was the Thesis Co-director of KF-S, participating in the writing of the manuscript and approving the final version of the manuscript to be submitted.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Centro de Investigación Biomédica del Sur (CIBIS)Instituto Mexicano del Seguro Social (IMSS)XochitepecMexico
  2. 2.Departamento de BiotecnologíaUniversidad Autónoma Metropolitana-IztapalapaCiudad de MéxicoMexico

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