Effect of Dichloromethane-Methanol Extract and Tomentin Obtained from Sphaeralcea angustifolia Cell Suspensions in a Model of Kaolin/Carrageenan-Induced Arthritis

Abstract

Cell suspension cultures from Sphaeralcea angustifolia produce compounds with anti-inflammatory activity, including scopoletin, tomentin, and sphaeralcic acid. Antiarthritic activity of scopoletin isolated from S. angustifolia and Erycibe obtusifolia wild plants was demonstrated in a complete Freud’s adjuvant-induced arthritis model. In this project, the dichloromethane-methanol extract and tomentin isolated from S. angustifolia cell suspension were evaluated in mice using a kaolin/carrageenan-induced arthritis model. After 9 days of treatment, the anti-inflammatory effect of S. angustifolia dichloromethane-methanol extract was 72% at a dose of 100 mg/kg, a recovery of 55% was observed with methotrexate (5.0 mg/kg), and the anti-inflammatory effect for tomentin was dose-dependent with a median effective dose of 10.32 mg/kg. The concentration of pro-inflammatory cytokines interleukin-1β and tumor necrosis factor alpha in the left joint of groups treated with S. angustifolia dichloromethane-methanol extract and tomentin at different doses was significantly lower than that detected in the group damaged with kaolin/carrageenan, and the concentration of interleukin-10 and interleukin-4 (anti-inflammatory cytokines) was significantly higher. The drop in mean body weight of mice treated with methotrexate (−4.34±0.47 g) was higher in comparison with the kaolin/carrageenan group (−2.40±0.29 g). On the other hand, the mean body weight of mice treated with S. angustifolia dichloromethane-methanol extract (−1.19±0.17 g) was similar, and mice treated with tomentin (0.19±0.11 g) was lower. The dichloromethane-methanol extract and tomentin isolated from S. angustifolia diminished joint edema induced by kaolin/carrageenan, possibly by acting as an immunomodulatory of the inflammatory response.

Production of anti‐inflammatory compounds in Sphaeralcea angustifolia cell suspension cultivated in stirred tank bioreactor

“…Maximal biomass of cell suspension in culture from S. angustifolia obtained in the bioreactor was 1.42-fold superior to that generated in the cultures developed in Erlenmeyer flasks [11]; nevertheless, the max was lowest. This result is consistent with that described for other species, such as B. vulgaris [13], A. indica [15], and Burcera linanoe [22]; maximal biomasses were obtained in the stirred tank-type bioreactor rather than in Erlenmeyer flasks.…”

“…Dichloromethane:methanol extract from biomass of cells in suspension of S. angustifolia, as well as tomentin, sphaeralcic acid, and scopoletin, were active as anti-inflammatories and reduced the mean body weight lost in Freund adjuvant-and kaolin/carrageenan-induced arthritis, respectively. In addition, all of them modulate the response of proinflammatory IL-1 , IL-6, and TNF-and anti-inflammatory IL-4 and IL-10, and scopoletin possess an antiangiogenic activity [10,11].…”

“…C during a light:dark (16 h:8 h) photoperiod under 50 M/m/s warm white fluorescent light intensity. The successful S. angustifolia cell suspension was transferred into a new medium under sterile conditions every 16days, utilizing the same inoculum of 4% w/v for a period of 2 years[9,11].…”

Pharmacokinetic Study of Anti-osteoarthritic Compounds of a Standardized Fraction from Sphaeralcea Angustifolia

“…Aliquots of 20 mL were obtained, tomentin was isolated from pooled sub-fraction of 80:20-H 2 O:CH 3 CN and sphaeralcic acid from 50:50-H 2 O:CH 3 CN. The compounds were analyzed by HPLC, and their purity was confirmed by comparing their retention times and absorption spectra [ 14 , 15 , 17 ]. This procedure was performed several times until the necessary amount of tomentin and sphaeralcic acid was obtained.…”

“…S. angustifolia has normative restriction [ 12 ] for its collection in its natural habitat; therefore, to obtain the phytopharmaceuticals responsible of the anti-osteoarthritis activity, the sustainable production of scopoletin, tomentin, and sphaeralcic acid ( Figure 1 ) in the cell suspension cultivated in a stirred tank bioreactor was implemented [ 13 ]. It should be highlighted that the active compounds significantly increased anti-inflammatory interleukin responses in rats with experimental arthritis [ 11 , 14 , 15 ]. Derived from the advances mentioned above, in the development of anti-osteoarthritic phytomedicine formulated with standardized Spheralceae angustifolia extract, and which may be of medical interest to drug developers, the aim of this research was to determine the plasma concentrations of scopoletin, tomentin, and sphaeralcic acid present in the standardized fraction (SaTSS) from an S. angustifolia cell after a single oral or intravenous dose in mice.…”