RESUMO
ABSTRACT Juglans regia L., Juglandaceae, is broadly used due to its immunomodulatory effects, potentials in protecting against many sever-disorders, and high safety-profile. The aim of this work is to make a phytochemical analysis of J. regia oil and kernel exploring their antinociceptive and anti-inflammatory potentials utilizing combined bio-guided gas chromatography with mass spectrometer (GC-MS), gas chromatography with flame ionization detection (GC-FID) and reversed-phase high-performance liquid chromatography (RP-HPLC) analyses. Combined bio-guided GC-MS, GC-FID and RP-HPLC analyses is an innovative-combined-technique aiming at efficiently analyzing various-extracts phytochemical and biological characters. The J. regia oil and kernel ethyl-acetate extract were monitored during exploring their possible acute-antinflammatory, antidiabetic and antidiabetic-neuropathy. Glycated-hemoglobin, serum-insulin, serum-catalase and lipid-peroxidation levels have been also monitored. Combined bio-guided GC-FID, GC-MS and HPLC analyses have shown to be an efficient analyzing-method through identifying the most active compound, linoleic acid. Linoleic acid has shown the highest improvement of the acute inflammatory-pain, chronic blood-glucose level reduction, serum-insulin elevation, and normalization of glycated-hemoglobin levels. J. regia oil has shown more lipid-peroxidation reduction, while kernel ethyl-acetate extract has shown more acute-blood-glucose level reduction and serum-catalase levels elevation. Compared to tramadol, the highest-doses of J. regia oil, kernel ethyl-acetate extract, and linoleic acid have shown higher antinociceptive-potentials in amelioration of thermal-hyperalgesic and anti-allodynic neuropathic-pain. Thus, the antinflammatory, the reduction of oxidative-stress, and the insulin-secretagogue potentials might be among the possible mechanisms of improvement of neuropathic-pain. In correlation to conventional-techniques, the combined bio-guided analyses have shown to be an efficient innovative-combined technique. After further clinical studies, J. regia might be utilized as a possible-remedy for various painful-syndromes.
RESUMO
Background: Bioactive compounds from the fruit of S. elaeagnifolium were isolated since could be highly potential source to develop functional foods or pharmaceutical products. Objectives: In this study a bioguided fractionation of the methanolic extract from S. elaeagnifolium fruit was carried out to evaluate their cytotoxicity and antitumoral potential on several cell lines and breast tumor explants, respectively. Methods: Microdilution method with A. salina was used to isolate bioactive compounds. Fractionation was performed by vacuum liquid chromatography, and the monitoring from fractions was done by thin layer chromatography. The effect of the fractions on viability in Vero, HeLa, and MCF-7 cells was assessed using WST-1 assay, whereas in breast tumor explants was evaluated by Alamar blue assay. A qualitative phytochemical analysis was performed to partially identify the compounds contained in the fractions and a spectroscopic characterization by RP-HPLC-MS was done to identify the group of compounds responsible for the effect on the cell lines and the mammary explants. Results: Several fractions were isolated from the fruit of S. elaeagnifolim. Notwithstanding, the FVLC7 showed a higher activity in A. salina assay. This fraction reduced the viability at 39 ± 1.67, 15.05 ± 0.09 and 66.10 ± 4 % in Vero, HeLa and MCF-7 cells, at 100 mg/mL, respectively. On the other hand, showed an effect in breast tumor explants obtained from a patient in remission. Qualitative phytochemical analysis showed that FVLC7 contains alkaloids, coumarins, and sesquiterpene lactones. Characterization by RP-HPLC-MS detected quinic acid, chlorogenic acid, dicaffeoylquinic acid as well as presence of an alkaloid. Conclusion: On this basis, our results suggest that cytotoxic effect of FVLC7 isolated from the fruit of S. elaeagnifolium could be mediated by quinic, chlorogenic, and dicaffeoylquinic acids
Antecedentes: Los compuestos bioactivos del fruto de S. elaeagnifolium fueron aislados ya que representan compuestos con un alto potencial para el desarrollo de alimentos funcionales o productos farmacéuticos. Objetivos: En este estudio se realizó un fraccionamiento biodirigido de un extracto metanólico de frutos de S. elaeagnifolium para evaluar la citotoxicidad y el potencial antitumoral en los explantes de tumor de mama. Métodos: Se utilizó el método de microdilución con A. salina para aislar los compuestos bioactivos. El fraccionamiento se realizó mediante cromatografía de líquido a vacío, y la monitorización de las fracciones se realizó por cromatografía en capa fina. La viabilidad de las fracciones en las líneas de células Vero, HeLa y MCF-7 se evaluó usando el ensayo WST-1, mientras que en los explantes tumorales de mama se evaluaron mediante el ensayo azul de Alamar. Así mismo, se realizó un análisis fitoquímico cualitativo para identificar parcialmente los compuestos que contenían las fracciones y una caracterización espectroscópica por RPHPLC-MS de los compuestos responsables del efecto sobre las líneas celulares y los explantes mamarios. Resultados: De todas las fracciones aisladas de S. elaeagnifolium, la fracción FVLC7 (100 µg/mL) tuvo la actividad más alta en el ensayo de A. salina. Por otra parte, redujo la viabilidad un 39 ± 1,67, 15,05 ± 0,09 y 66,10 ± 4,44% en las células Vero, HeLa y MCF-7, respectivamente. Esta fracción mostró un efecto en los explantes de tumor de mama obtenidos de un paciente en remisión. El análisis fitoquímico cualitativo reveló que la FVLC7 contiene alcaloides, cumarinas y lactonas sesquiterpénicas. La caracterización por RP-HPLC-MS detectó ácido quínico, ácido clorogénico, ácido dicafeoilquínico así como presencia de un alcaloide. Conclusión: Nuestros resultados sugieren que el efecto tóxico de la fracción FVLC7 aislada del fruto de S. elaeagnifolium podría deberse a los ácidos quínico, clorogénico y dicafeoilquínico.