An Ethanolic Extract of Boehmeria Caudata Aerial Parts Displays Anti-Inflammatory and Anti-Tumor Activities
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Published online: 2020-03-18 Original Papers Thieme An Ethanolic Extract of Boehmeria caudata Aerial Parts Displays Anti-inflammatory and Anti-tumor Activities Authors Paula Pereira de Paiva1, 2, 3 , Fabiana Regina Nonato2, Ana Lúcia Tasca Gois Ruiz3 , Ilza Maria de Oliveira Sousa2, 3, Rafael Rosolen Teixeira Zafred1, 2, Diogo Noin de Oliveira3, Rodrigo Ramos Catharino3, Mary Ann Foglio3, João Ernesto de Carvalho3 Affiliations 1 Institute of Biology, University of Campinas, Campinas, Supporting Information for this article is available online Brazil at http://www.thieme-connect.de/products. 2 Chemical, Biological and Agricultural Research Center, (CPQBA), University of Campinas, Paulínia, Brazil ABstract 3 Faculty of Pharmaceutical Sciences, University of The tumor microenvironment presents several therapeutic Campinas, Campinas, Brazil targets, with inflammation being one of them. In search of new drugs, plants have shown to be an effective source of potent Key words anti-inflammatory and anticancer agents. This study aimed to Boehmeria caudate, Urticaceae, inflammation, antineoplas- evaluate the antitumoral and inflammatory activities ofBoe - tic agents, Ehrlich tumor carcinoma, drug screening assays hmeria caudata aerial parts extract. Bioguided in vitro antipro- liferative screening showed that phenanthroquinolizidine ob- received 20.11.2019 tained from the aerial B. caudata ethanolic extract had a straight 18.01.2020 revised relationship with activity. Moreover, the orally administered accepted 22.01.2020 ethanolic extract reduced Ehrlich solid tumor growth and dis- played an anti-inflammatory effect in both evaluated experi- Bibliography mental models (carrageenan-induced paw edema and croton DOI https://doi.org/10.1055/a-1111-9907 oil-induced ear edema). These results suggest that the antitu- Planta Med Int Open 2020; 7: e17–e25 mor activity of the ethanolic extract could be explained by © Georg Thieme Verlag KG Stuttgart · New York antiproliferative effects associated with anti-inflammatory ac- ISSN 2509-9264 tion. Correspondence: Dra. Paula Pereira de Paiva Faculty of Pharmaceutical Sciences, University of Campinas, Cândido Portinari, 200 13083-871, Cidade Universitária Zeferino Vaz Campinas SP Brazil Tel.: + 55 (19) 35217715 [email protected] de Paiva PP et al. An Ethanolic Extract of … Planta Med Int Open 2020; 7: e17–e25 e17 Original Papers Thieme were classified as active when TGI ≤ 50 μg mL − 1 against at least two tumor cell lines in a panel of three tumor cell lines [15]. DE weakly ABBreViations inhibited breast (MCF-7, TGI: 35.26 μg mL − 1) and lung (NCI-H460, AE: aqueous extract TGI: 38.19 μg mL − 1) tumor cell lines while AE showed weak and AEF: alkaloid enriched fraction moderate activity on glioma (U251, TGI: 27.73 μg mL − 1) and lung CG: carrageenan (NCI-H460, TGI: 6.67 μg mL − 1) tumor cell lines in the order given. COX: cyclooxygenase The most active samples were EE and AEF, which potently inhibit- DE: dichloromethane extract ed (TGI < 0.25 μg mL − 1) the three tumor cell lines. Further, all ex- EE: ethanolic extract tracts inhibited the non-tumor cell line HaCat (▶Table 1). HaCat: immortal keratinocyte non-tumor human line Using TLC with Dragendorff reagent (▶Fig. 1a–m), the expect- i.p.: intraperitoneal able presence of alkaloids was confirmed in theB. caudata aerial LOX: lipoxygenase parts ethanolic extract by the regard of fluorescent spots under UV, MCF-7: breast human tumor cell line which were brownish due to the Dragendorff reagent. Acid-based NCI-H460: non-small cell lung human tumor line extraction of EE afforded the AEF (▶Fig. 1j–m). Thus, the presence o.r.: oral of alkaloids in both EE and AEF could explain the alikeness in the RTW: relative tumor weight antiproliferative profile of these samples. TPA: 12-O-tetracanoylphorbol lipoxygenase-13-ac- High-resolution electrospray ionization mass spectrometry (HRE- etate SI-MS) analysis of AEF allowed for the putative identification and rel- TGI: total growth inhibition ative quantification of five phenanthroquinolizidine alkaloids (cryp- t.t.: topical treatment topleurine, boehmeriasin A, boehmeriasin B, julandine, hydroxycryp- U251: glioma human tumor cell line topleurine), one quinolizidine alkaloid [3-(4-hydroxyphenyl)-4- (3-methoxy-4-hydroxyphenyl)-3,4-dehydroquinolizidine], and two acetophenone alkaloids (3,4-dimethoxy-ω-(2'-piperidy1) aceto- phenone and 2’,4’dimethoxyacetofenone) (▶Table 2). Except for cryptopleurine, seven alkaloids were identified for the first time in Introduction B. caudata aerial parts extract. Considered one of the hallmarks of cancer, inflammation substan- Due to the antiproliferative effect of EE, the anti-inflammatory tially contributes to the development and progression of malignan- [18] and antitumor [8, 11, 18] activities of the identifiable alkaloids cies [1]. Many plant-derived natural products have been described (▶Table 2), and the availability of test material, further assess- as potent anti-inflammatory and anticancer agents, in which they ments were conducted on in vivo antitumor and anti-inflammato- provide alternative strategies for the development of new drugs ry models to establish the in vivo correlation between the two ac- [2–4]. tivities. Boehmeria caudata Sw (Urticaceae) aerial parts extract had showed Following the acute toxicity protocol [19], the higher adminis- a promising antiproliferative effect (total growth inhibition lower trated dose of EE that promotes no clinical adverse effects was de- than 0.25 μg mL − 1) in a preliminary screening done by our research termined using two different routes of administration. Neverthe- team [5]. This species is widely spread across the neotropical re- gion, that is, throughout Central and South America [6]. Found in ▶Table 1 B. caudata extracts in vitro antiproliferative activity (TGI, μg the Southeast and Southern regions, and in the State of Mato Gros- mL − 1). so do Sul in Brazil, B. caudata is well known as “lixa-da-folha-larga” and “assa-peixe” [7]. Cell linesa Despite the traditional usage, there is only one study aiming at Sam- U251 MCF-7 NCI-H460 HaCat the investigation of the pharmacological potential of B. caudata ples that describes the cytotoxic activity of B. caudata stem wood etha- DE 90.50 I 35.26 W 38.19 W 35.57 W nolic extract upon human epidermoid carcinoma of the nasophar- EE < 0.25 P < 0.25 P 0.66 P < 0.25 P ynx [8]. Considering the Boehmeria genus, there is evidence of an- AE 27.73 W 63.57 I 6.67 M 7.94 M ti-inflammatory and antitumor effects in addition to the presence AEF < 0.25 P < 0.25 P 0.66 P < 0.25 P of phenanthroquinolizidine alkaloids, such as cryptopleurine, in DOXO 2.66 P 3.69 P > 25 W 19.41 W some extracts [9–14]. Therefore, the present study prompted the TGI: total growth inhibition after 48 h exposition. TGI values were in vitro and in vivo evaluation of anti-inflammatory and antitumor calculated by nonlinear regression analysis using ORIGIN 8.0 effects of the ethanolic extract ofB. caudata aerial parts. (OriginLab Corporation). aHuman tumor cell lines: U251 (glioma), MCF-7 (breast), NCI-H460 (lung, non-small cells); human non-tumor line: HaCat (immortal keratinocyte). DE: dichloromethane extract, Results and Discussion EE: ethanolic extract, AE: aqueous extract; AEF: alkaloid enriched fraction; DOXO: doxorubicin (positive control). CSIR’s criteria: DE, EE, and AE of B. caudata aerial parts and AEF were evaluated on inactive (I, TGI > 50 μg mL − 1), weak activity (W, 15 μg four human cell lines (one non-tumor and three tumor cell lines) mL − 1 < TGI < 50 μg mL − 1), moderate activity (M, 6.25 μg − 1 − 1 − 1 using doxorubicin as a positive control. Expressed as the sample mL < TGI < 15 μg mL ), and potent activity (P, TGI < 6.25 μg mL ) [15]. concentration required for TGI (μg mL − 1), the extracts and fraction e18 de Paiva PP et al. An Ethanolic Extract of … Planta Med Int Open 2020; 7: e17–e25 AB CDEF GH IJLM ▶Fig. 1 Thin-layer chromatography plates (silica gel 60 F254) of dichloromethane (DE, a–c), ethanolic (EE , d–e), aqueous (AE, g–i) extracts of Boehmeria caudata aerial parts and alkaloid rich fraction (AEF, j–m). Stationary phase: Mobile phase: dichloromethane/methanol (97:3, for DE) and butanol/acetic acid/distilled water (4:1:5, for EE, AE and AEF). Detection: under UV light (254 nm: D, G and L; 366 nm: A, E, H and J); Anisaldehyde (B) and Dragendorff (C, F, I and M) solutions. less, when administrated via the i.p. route, EE (500 mg kg − 1) pro- ed a reduction of 33.88 % (3 mg kg − 1, RTW = 0.00199 ± 0.0003, moted depression, piloerection, tachypnea, and palpebral ptosis p < 0.05. There was no statistically significant difference among the up to 4 h after administration, which suggested central nervous treated groups (▶Fig. 2). Furthermore, EE doses did not produce system effects. After 24 h, the EE-treated mice showed normal be- any clinical signs of toxicity during treatment. havior, which persisted throughout the experiment. In addition, at It is well known that Ehrlich cell growth generates a local a higher dose (1000 mg kg − 1, i.p.), EE promoted signs and symp- inflammatory response characterized by increased vascular perme- toms of central nervous system depression, which resulted in the ability, edema formation, cell migration, and recruitment of the death of mice within the first 24 h. Based on these results, we chose immune response [21]. As we take the tumor microenvironment 150 mg kg − 1 (30 % of the lower toxic dose on acute evaluation) of into consideration as a therapeutic target, EE was evaluated by two EE as the highest dose on the pharmacological experiments using different inflammation models in mice. Since i.p. administration the i.p. route. can induce the local inflammatory process, the o.r. route was cho- In the same manner, after oral treatment, EE at 1000 mg kg − 1 sen for these assays.