Evaluation of the antigenotoxic and antioxidant activity induced by Croton antisyphiliticus

SILVA, REGILDO M.G. DA; FIGUEIREDO, CÉLIA C.M.; GOMES, AMANDA C.; FERREIRA, PAULO C.; GRANERO, FILIPE O.; SANTIAGO, PATRÍCIA S.; SILVA, LUCIANA P.

doi:10.1590/0001-3765202020190047

REGILDO M.G. DA SILVA

Universidade Estadual Paulista/UNESP, Faculdade de Ciências e Letras de Assis, Departamento de Biotecnologia, Laboratório de Plantas Medicinais e Produtos Naturais, Av. Dom Antônio, 2100, Parque Universitário, 19806-900 Assis, SP, Brazil

https://orcid.org/0000-0002-0300-0247

CÉLIA C.M. FIGUEIREDO

Universidade Estadual Paulista/UNESP, Faculdade de Ciências e Letras de Assis, Departamento de Biotecnologia, Laboratório de Plantas Medicinais e Produtos Naturais, Av. Dom Antônio, 2100, Parque Universitário, 19806-900 Assis, SP, Brazil

https://orcid.org/0000-0003-0449-9200

AMANDA C. GOMES

Universidade Estadual Paulista/UNESP, Faculdade de Ciências e Letras de Assis, Departamento de Biotecnologia, Laboratório de Plantas Medicinais e Produtos Naturais, Av. Dom Antônio, 2100, Parque Universitário, 19806-900 Assis, SP, Brazil

https://orcid.org/0000-0002-6576-5536

PAULO C. FERREIRA

Universidade Estadual Paulista/UNESP, Faculdade de Ciências e Letras de Assis, Departamento de Biotecnologia, Laboratório de Plantas Medicinais e Produtos Naturais, Av. Dom Antônio, 2100, Parque Universitário, 19806-900 Assis, SP, Brazil

https://orcid.org/0000-0001-8133-6451

FILIPE O. GRANERO

Universidade Estadual Paulista/UNESP, Faculdade de Ciências e Letras de Assis, Departamento de Biotecnologia, Laboratório de Plantas Medicinais e Produtos Naturais, Av. Dom Antônio, 2100, Parque Universitário, 19806-900 Assis, SP, Brazil

https://orcid.org/0000-0002-2803-4267

PATRÍCIA S. SANTIAGO

Universidade Estadual Paulista/UNESP, Campus de Registro, Av. Nelson Brihi Badur, 430, Vila Tupy, 11900-000 Registro, SP, Brazil

https://orcid.org/0000-0002-6205-9441

LUCIANA P. SILVA

Fundação Educacional do Município de Assis/FEMA, Av. Getulio Vargas, 1200, Vila Nova Santana, 19807-130 Assis, SP, Brazil

https://orcid.org/0000-0002-0914-2094

Correspondence to: Regildo Márcio Gonçalves da Silva E-mail: regildos@yahoo.com.br

AUTHOR CONTRIBUTIONS

R.M.G. Silva performed all the phytochemical analysis, fractionation and isolation of the compounds active. C.C.M. Figueiredo, A.C. Gomes and P.S. Santiago performed antioxidants tests. F.O. Granero performed collecting the plant material and help in writing the manuscript. P.C. Ferreira performed the antigenotoxic assays. This work was done under the supervision of L.P. Silva and R.M.G. Silva. The authors declare that they have no conflict of interest.

Figures (2)
Tables (5)

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Figure 1
HPLC fingerprints obtained by methanolic extract (a), ethyl acetate fractions (b) and standard compounds (c): 1-kaempferol, 2-orientin, 3-rutin, 4-vitexin, 5-quercitin and 6-myricetin.

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Figure 2
Chemical structure the vitexin of Croton antisyphiliticus Mart.

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Table I
Mitotic Index and Micronucleated polychromatic erythrocyte (MNPCE) frequency in Swiss albino mice (n=5) treated with different concentrations of methanolic extract (ME) (1, 3 and 6 mg 100g^-1bw) and fractions (N-Hexane=Hex, Dichloromethane = CH₂Cl₂, Ethyl Acetate = AcOEt, Methanol = MeOH) (6.0 mg g^-1bw), vitexin isolated from C. antisyphiliticus (Vitexin_ca) (1.0 mg g^-1bw), standard sample vitexin (Vitexin_ss) (1.0 mg g^-1bw), control group (CG) and group (CP) treated with Cyclophosphamide® (2mg 100g^-1bw). 2000 Polychromatic Erythrocytes (PCE) per mouse.

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Table II
Mitotic Index and Micronucleated polychromatic erythrocyte (MNPCE) frequency in Swiss albino mice (n=5) treated with different concentrations of methanolic extract (ME) (1, 3 and 6 mg 100g^-1bw) and fraction (Ethyl Acetate = AcOEt) (6.0 mg g^-1bw), vitexin isolated from C. antisyphiliticus (Vitexin_ca) (1.0 mg g^-1bw), standard sample vitexin (Vitexin_ss) (1.0 mg g^-1bw) in association with Cyclophosphamide® (2mg 100g^-1bw), control group (CG) and group (CP) treated with Cyclophosphamide® (2mg 100g^-1bw). 2000 Polychromatic Erythrocytes (PCE) per mouse.

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Table III
DPPH radical scavenging activity, iron ion chelating power, inhibition of lipid peroxidation (TBARS), NO radical scavenging and determination of total phenols and flavonoids at different concentration of methanolic extract (ME) of C. antisyphiliticus.

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Table IV
DPPH radical scavenging activity and determination of total content of phenols and flavonoids in different fractions (N-Hexane=Hex, Dichloromethane = CH₂Cl₂, Ethyl Acetate = AcOEt, Methanol = MeOH) of methanolic extract of C. antisyphiliticus at concentration of 500 µg mL^-1).

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Table V
DPPH radical scavenging activity of vitexin isolated from C. antisyphiliticus (Vitexin_ca), standard sample vitexin (Vitexin_ss) and positive control (Rutin and Caffeic acid).

Groups	mg 100g^-1bw	%MNPCE±DP	Mitotic Index
CG	0.0	0.38 ± 0.05a	10.38±0.52a
Extract ME
	1,0	0.27 ± 0.06b	9.72±0.23a
	3.0	0.23 ± 0.05b	9.01±0.39a
	6.0	0.18 ± 0.02b	9.23±0.87a
Fractions
Hex	6.0	0.35±0.05a	10.33±1.09a
CH₂Cl₂	6.0	0.27±0.09b	10.21±1.17a
AcOEt	6.0	0.04±0.02c	9.98±0.98a
AcOEt:MeOH (70:30)	6.0	0.18±0.04b	10.78±1.09a
MeOH	6.0	0.41±0.04a	10.17±1.12a
Vitexin_ca	1.0	0.02±0.001c	10.08±0.18a
Vitexin_ss	1.0	0.31±0.09b	9.78±0.79a
CP	2.0	2.85 ± 0.38d	4.82±0.81b

Groups	mg 100g^-1bw		%MNPCE±DP	Mitotic Index
Groups	Treatments	Cyclophosphamide	%MNPCE±DP	Mitotic Index
CG	0.0	0.0	0.34 ± 0.09a	11.17±0.61a
ME+CP	1.0	2.0	1.78 ± 0.23b	7.24±0.19b
	3.0	2.0	1.03 ± 0.13b	8.01±0.11b
	6.0	2.0	0.98 ± 0.17b	8.73±0.27b
AcOEt+CP	6.0	2.0	0.31 ± 0.08a	10.18±0.44a
Vitexin_ca+CP	1.0	2.0	0.16 ± 0.01c	10.89±0.31a
Vitexin_ss+CP	1.0	2.0	1.77 ± 0.19b	9.17±0.43b
CP	0.0	2.0	3.17 ± 0.56d	5.42±0.32c

ME concentrations (µg mL^-1)	% DPPH radical scavenging^a	% Iron ion chelate	TBARS- Inhibitory activity (%)	% NO radical scavenging	Total phenol contents^b	Flavonoid contents^c
100	22.1±1.22a	31.40±1.01a	38.65±7.05a	51.81±3.54a	0.98±0.01	1.71±0.02
250	26.2±1.03a	40.30±2.33a	39.02±9.90a	43.67±2.18a	2.25±0.17	1.89±0.03
500	71.8±2.09b	53.00±2.89b	39.88±7.33a	53.78±0.36a	4.05±0.09	2.33±0.11
1000	73.0±2.78b	64.23±1.98b	37.76±10.6a	70.91±0.99b	7.06±0.98	3.24±0.05
1500	79.4±3.09bc	72.15±4.09bc	45.77±2.65a	74.20±0.60b	10.32±1.11	3.89±0.97
2000	81.0±3.23c	77.80±3.93c	32.49±7.17a	80.97±0.81b	14.54±1.27	4.01±0.83

Fractions (500 µg mL^-1)	% DPPH radical scavenging^a	Total phenol contents^b	Flavonoid contents^c
Hex	-	-	-
CH₂Cl₂	17.14±0.21a	9.09±0.99a	5.44±0.18a
AcOEt	65.46±1.22b	66.40±1.98b	59.42±1.49b
AcOEt:MeOH (70:30)	49.17±2.09c	43.34±1.79c	40.26±1.17c
MeOH	34.25±1.32d	23.28±1.33d	19.10±0.98d

Compound	DPPH - IC₅₀ (µg/mL)
Vitexin_ca	10.49±0.11a
Vitexin_ss	16.12±0.14b
Rutin	11.23±0.22a
Caffeic acid	15.27±0.21b

[1] Correspondence to: Regildo Márcio Gonçalves da Silva E-mail: regildos@yahoo.com.br

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Evaluation of the antigenotoxic and antioxidant activity induced by Croton antisyphiliticus

Abstract