Thrombolysis, clotting, and genotoxic activities modulated by essential oils extracted from <i>Lippia alba</i>

Espósito, Mariana Araújo; Trento, Marcus Vinicius Cardoso; Braga, Mariana Aparecida; Marcussi, Silvana

doi:10.1590/s2175-97902022000X2e20575

Mariana Araújo Espósito

Biochemistry Laboratory, Department of Chemistry, Federal University of Lavras, Lavras, Minas Gerais, Brazil

Marcus Vinicius Cardoso Trento

Biochemistry Laboratory, Department of Chemistry, Federal University of Lavras, Lavras, Minas Gerais, Brazil

Mariana Aparecida Braga ^**Correspondence: M. A. Braga. Laboratório de Bioquímica. Departmento de Química. Universidade Federal de Lavras. Campus Universitário, CP: 3037. Lavras 37200-900, Brasil. Phone: +55 (35)3829-1893. E-mail: marybraga07@yahoo.com.br.

Biochemistry Laboratory, Department of Chemistry, Federal University of Lavras, Lavras, Minas Gerais, Brazil

https://orcid.org/0000-0003-0758-3993

Silvana Marcussi

Biochemistry Laboratory, Department of Chemistry, Federal University of Lavras, Lavras, Minas Gerais, Brazil

https://orcid.org/0000-0002-4674-6911

*Correspondence: M. A. Braga. Laboratório de Bioquímica. Departmento de Química. Universidade Federal de Lavras. Campus Universitário, CP: 3037. Lavras 37200-900, Brasil. Phone: +55 (35)3829-1893. E-mail: marybraga07@yahoo.com.br.

CONFLICT OF INTEREST

The authors declare that they have no conflict of interest.

Figures (3)
Tables (3)

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FIGURE 1
Activity on human blood thrombi. (A) Thrombolytic activity of essential oils from L. alba flower and leaf and activity induced by B. moojeni venom. (B) Effect of essential oils on the thrombolytic activity induced by L. muta muta venom. (+)C: Positive Control: 40 µg mL^-1 of B. moojeni or L. muta muta venom; (-)C: Negative Control: PBS; L. alba flower and leaf essential oil (F and L; 1.2 µL mL^-1). Incubated: Venoms + essential oils at different ratios. Each letter statistically differs (p < 0, 05) using Skott-Knott test.

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FIGURE 2
Effect of L. alba flower and Leaf EO on DNA from human blood leukocytes. Percentage of nucleoids in the different comet classes corresponding to the varying degradation levels of the DNA molecules. Data represent average of values obtained from triplicate assays where were evaluated 300 nucleoids/treatment/assay. Comets were classified according to Collins et al. (1997Collins A, Dušinská M, Franklin M, Somorovská M, Petrovská H, Duthie S, et al. Comet assay in human biomonitoring studies: Reliability, validation, and applications. Environ Mol Mutagen. 1997;30(2):139-146.). NC: negative control (PBS); +CD: positive control (Doxorrubicin, 100 µg mL^-1); +CL: positive control (L. muta muta venom, 50 µg mL^-1). Oil + Venom (L. muta muta venom). F: flower oil. L: leave oil. Volume of oil in μL.

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FIGURE 3
Comet assay (single cell gel electrophoresis) in human blood leukocytes. Demonstrative images of scores observed in different Damage Classes (form 0 to 4). (+)CL: Positive Control with L. muta muta, 50 µg mL^-1; (+) CD: Positive Control with Doxorubicin, 100 µg mL^-1.

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TABLE I
Chemical constituents of the essential oils of flowers and leaves of L. alba

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TABLE II
Effect of essential oils from L. alba on clotting time of human citrated plasma induced by B. moojeni and L. muta muta venoms

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TABLE III
Effect of L. alba EO on DNA from human blood leukocytes

Essential oil %	Compound	RI
Flower
9.1	ρ-Cimene	1026
0.06	Limonene	1029
5.16	γ-Terpinene	1058
32.75	Neral	1239
49.83	Geranial	1269
2.07	D-Germacrene	1481
1.03	α-Elemol	1550
Leaf
0.13	α-Thujeno	924
1	Sabinene	972
*	ρ-Cimene	1.025
18.03	Limonene	1.028
5.02	γ-Terpinene	1.057
31.02	Neral	1.238
38.06	Geranial	1.268
3.17	D- Germacrene	1.480
3.52	α -Elemol	1.549

*Samples L. alba* oil (µL mL** ^-1 )		Clotting time (s)
*Samples L. alba* oil (µL mL** ^-1 )		B. moojeni venom	L. muta muta venom
Control (without oil)		66.66 ± 2.1	61.33 ± 2.1
Leaf	0.05	78.6 ± 1.5^a	74.6 ± 2.3^a
	0.1	47.6 ± 1.2^b	90.3 ± 2.1^a
	0.3	44.0 ± 2.0^b	92.6 ± 0.6^a
	0.6	45.6 ± 1.5^b	110.0 ± 2.0^a
	1.2	44.3 ± 0.6^b	113.0 ± 1.0^a
Flower	0.05	57.0 ± 1.2 *	80.0 ± 0.6 ^a
	0.1	65.6 ± 1.2 *	65.9 ±1.5*
	0.3	57.0 ± 1.7 *	65.9 ±0.6*
	0.6	52.3 ± 2.5^b	73.6 ±2.9^a
	1.2	46.3 ± 2.5^b	83.6 ±2.3^a

Amount (µL mL^-1)	Samples	Damage frequency (%)	Arbitrary units; A ± SD
Amount (µL mL^-1)	(-) C	7.1	28.3 ± 8.02^a
100 µg mL^-1	(+) CD	80.3	321.3 ± 45.4^b
50 µg mL^-1	(+) CL	70.7	282.7 ± 33.8^b
1.5	Flower EO	14.4	57.8 ± 6.15^c
3.0		13.3	53.3 ± 2.31^c
6.0		15.4	61.7 ± 3.51^c
1.5	Leaf EO	7.7	30.7 ± 4.16^a
3.0		7.6	30.0 ± 3.60^a
6.0		8.4	33.0 ± 5.6^a
1.5	L. muta muta + Flower EO	58.8	235.0 ± 11.31^b
3.0	L. muta muta + Flower EO	26.8	107.0 ± 4.24^d
1.5	L. muta muta + Leaf EO	14.5	58.0 ± 1.41^c
3.0	L. muta muta + Leaf EO	11.6	46.5 ± 6.36^c

[1] *Correspondence: M. A. Braga. Laboratório de Bioquímica. Departmento de Química. Universidade Federal de Lavras. Campus Universitário, CP: 3037. Lavras 37200-900, Brasil. Phone: +55 (35)3829-1893. E-mail: marybraga07@yahoo.com.br.

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Thrombolysis, clotting, and genotoxic activities modulated by essential oils extracted from Lippia alba

Abstract