Abstract
Sphaeralcea angustifolia (Cav) G. Don is used in traditional Mexican medicine to treat inflammations and gastric disease. Its anti-inflammatory and immunomodulatory activities in mice and rats acute and chronic models have been attributed mainly to scopoletin. Scopoletin reduced joint inflammation, the number of new vessels, production of endogenous angiogenic inducers, and reversed the histopathological alterations in rat adjuvant-induced arthritis. Tomentin and sphaeralcic acid from S. angustifolia cells in suspension proved anti-inflammatory and immunomodulatory activities in mice kaolin/λ-carrageenan-induced arthritis. Transformed roots of S. angustifolia have been proposed as active compounds producers. A high transformation frequency mediated by Agrobacterium rhizogenes ATCC15834/pTDT was obtained from nodal segments (59.5 ± 10.5%, 145 hairy root lines) and leaves (40.0 ± 25, 52 hairy root lines) of 2-month-old plantlets. Among seven lines selected according to their phenotypic characteristics and growth index, the SaTR N7.2 line presented the highest sphaeralcic acid production (17.6 ± 1.72 mg/g DW); this production was 440-fold superior to that reported in S. angustifolia wild plants, and in comparison to cells in suspension of S. angustifolia in MS medium with nitrate restriction this was 263-fold higher when cultured in flasks and 5-fold higher in a stirred-tank type bioreactor. The SaTR N7.2, SaTR N5.1, SaTR N7.1, and SaTR N15.1 lines excreted sphaeralcic acid into the culture medium at similar levels. Genetic transformation of hairy roots was confirmed by amplifying a 490 bp fragment of the rolC gene. After 2 years in culture, S. angustifolia hairy roots are producers of scopoletin and sphaeralcic acid and they can be stressed by nitrate reduction and/or copper increased to stimulate scopoletin and sphaeralcic acid production.
Highlights
IntroductionDon (Malvaceae) is known in Mexico as “Vara de San José” or “Hierba del negro” (Aguilar et al, 1994)
A high transformation frequency mediated by Agrobacterium rhizogenes ATCC15834/pTDT was obtained from nodal segments (59.5 ± 10.5%, 145 hairy root lines) and leaves (40.0 ± 25, 52 hairy root lines) of 2-month-old plantlets
Among seven lines selected according to their phenotypic characteristics and growth index, the SaTR N7.2 line presented the highest sphaeralcic acid production (17.6 ± 1.72 mg/g dry weight (DW)); this production was 440-fold superior to that reported in S. angustifolia wild plants, and in comparison to cells in the suspension of S. angustifolia in Murashige and Skoog (MS) medium with nitrate restriction this was 263-fold higher when cultured in flasks and 5-fold higher in a stirred-tank type bioreactor
Summary
Don (Malvaceae) is known in Mexico as “Vara de San José” or “Hierba del negro” (Aguilar et al, 1994). This plant is used in Mexican traditional medicine to treat blows, fractures, inflammation, and gastric problems (Aguilar et al, 1994; Argueta et al, 1994). Extracts from aerial parts of this plant had demonstrated anti-inflammatory activity, with scopoletin identified as the main active compound (García-Rodríguez et al, 2012; Juárez-Ciriaco et al, 2008; Meckes et al, 2004). Topical administration of a gel formulation made with 1% of S. angustifolia dichloromethane extract standardized in the scopoletin content showed therapeutic effectiveness and tolerability in patients with osteoarthritis of hands, capable of reducing the associated symptoms: pain, inflammation, and joint stiffness (Romero-Cerecero et al, 2013). Scopoletin has been isolated from many plants (Jain et al, 2002), and several biological activities had been reported, such as anti-inflammatory (Ding et al, 2008; Moon et al, 2007), antioxidant (Gwak et al, 2011; Shaw et al, 2003), anti-proliferative (Thani et al, 2010), anti-angiogenic (Pan et al, 2009), antithyroid, anti-hypertensive (Aldi et al, 2015), anti-hyperuricemic (Zeng et al, 2020), and anti-diabetic (Jang et al, 2020), among others
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