Exopolysaccharides from <i>Klebsiella oxytoca</i>: anti-inflammatory activity

Bannwart, Thays Avelino; Bersani-Amado, Ciomar Aparecida; Ames, Franciele Queiroz; Siqueira, Vera Lúcia Dias; Oliveira, Arildo José Braz de; Gonçalves, Regina Aparecida Correia

doi:10.1590/s2175-97902022e190511

Thays Avelino Bannwart

Department of Pharmacy, Graduate Program in Pharmaceutical Sciences, State University of Maringá, 87.020-900 Maringá, PR, Brazil

Ciomar Aparecida Bersani-Amado

Department of Pharmacology and Therapeutic, State University of Maringá, 87.020-900 Maringá, PR, Brazil

Franciele Queiroz Ames

Department of Pharmacology and Therapeutic, State University of Maringá, 87.020-900 Maringá, PR, Brazil

Vera Lúcia Dias Siqueira

Department of Clinical Analysis and Biomedicine, State University of Maringá, Maringá, PR, Brazil

Arildo José Braz de Oliveira

Department of Pharmacy, Graduate Program in Pharmaceutical Sciences, State University of Maringá, 87.020-900 Maringá, PR, Brazil

Regina Aparecida Correia Gonçalves ^**Correspondence: R. A. C. Gonçalves. Departamento de Farmácia. Universidade Estadual de Maringá. Avenida Colombo 5790, Maringá, Paraná, Brasil. Fone: +55-44-30115967. Email: racgoncalves@uem.br

Department of Pharmacy, Graduate Program in Pharmaceutical Sciences, State University of Maringá, 87.020-900 Maringá, PR, Brazil

http://orcid.org/0000-0003-4070-1269

*Correspondence: R. A. C. Gonçalves. Departamento de Farmácia. Universidade Estadual de Maringá. Avenida Colombo 5790, Maringá, Paraná, Brasil. Fone: +55-44-30115967. Email: racgoncalves@uem.br

CONFLICT OF INTEREST

The authors declare that there are no conflicts of interest. All authors have contributed equally.

Figures (3)
Tables (4)

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FIGURE 1
NMR spectra of the KEPS from K. oxytoca in D₂O: (A) ¹H NMR spectrum (300.06 MHz); (B) Zoom into anomeric region of the ¹H NMR spectrum; (C) ¹³C NMR spectrum (75.45 MHz).

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FIGURE 2
KEPS and KEPSC effects on the volume of pleural inﬂammatory exudate. The pleurisy was induced by intrapleural carrageenan injection (Cg - 200 µg/cavity) in rats (n = 6-7), 1 h following oral administration of the EPS at 400 mg/kg or saline (Cg). Indomethacin (Indo, 5 mg/kg) was orally administered as an anti-inflammatory reference drug (positive control). N = normal animals that received no pleural injection of carrageenan. Each point represents the mean ± standard error of the mean of volume of exudate 4 h after injection of carrageenan. ^a p < 0.05 when compared with normal rats; ^b p < 0.05 when compared with group of rats injected with carrageenan and untreated (Cg) (ANOVA, Tukey’s test).

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FIGURE 3
KEPS effect on the volume of pleural inﬂammatory exudate. The pleurisy was induced by intrapleural carrageenan injection (Cg - 200 µg/cavity) in rats (n = 6-7), 1 h following subcutaneous administration of the EPS at 50, 100, 200, 400 mg/kg or saline (Cg). Indomethacin (Indo, 5 mg/kg) was subcutaneously administered as an anti-inflammatory reference drug (positive control). N = normal animals that received no pleural injection of carrageenan. Each point represents the mean ± standard error of the mean of the volume of exudate 4 h after injection of carrageenan. ^a p < 0.05 when compared with normal rats; ^b p < 0.05 when compared with group of rats injected with carrageenan and untreated (Cg) (ANOVA, Tukey’s test).

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TABLE I
Total carbohydrate, reducing sugars, uronic acid, pyruvic acid, and protein content of exopolysaccharides from K. oxytoca

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TABLE II
¹³C (75.45 MHz, D₂O) NMR chemical shifts of KEPS from K. oxytoca

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TABLE III
Effect of oral administration of KEPS and KEPSC on the number of total and differential leukocytes in pleural inflammatory exudate of rats

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TABLE IV
KEPS effect on the number of total and differential leukocytes in pleural inflammatory exudate of rats

Sample	Total carbohydrate (%, w/w)	Reducing sugars (%, w/w)	Uronic acid (%, w/w)	Pyruvic acid (%, w/w)	Protein (%, w/w)
KEPSC	65.57^a	6.68^a	7.90^a	3.01^a	16.83^b
KEPS	62.82^b	6.50^b,a	6.21^a	1.68^b	8.92^c

¹³ C Chemical shifts (ppm)
Position	C-1	C-2	C-3	C-4	C-5	C-6
2-α-Rha	101.19 (100.3)	78.16 (78.0)	69.98 (69.7)	72.36 (71.9)	69.18 (69.1)	16.69 (16.5)
A	101.19 (100.3)	78.16 (78.0)	69.98 (69.7)	72.36 (71.9)	69.18 (69.1)	16.69 (16.5)
2-α-Rha	98.19 (99.5)	79.32 (79.0)	69.80 (69.8)	-	-	-
B	98.19 (99.5)	79.32 (79.0)	69.80 (69.8)	-	-	-
3,4-α-Rha	101.57 (101.9)	70.25 (69.6)	80.15 (80.6)	78.82 (78.2)	-	16.83 (16.9)
D	101.57 (101.9)	70.25 (69.6)	80.15 (80.6)	78.82 (78.2)	-	16.83 (16.9)
3-α-Rha	101.43 (101.9)	69.43 (69.7)	-	71.21 (71.1)	69.18 (69.2)	-
C	101.43 (101.9)	69.43 (69.7)	-	71.21 (71.1)	69.18 (69.2)	-
t-β-GlcA	102.33 (102.4)	72.88 (73.4)	75.13 (75.7)	72.03 (72.0)	76.96 (76.5)	-
E	102.33 (102.4)	72.88 (73.4)	75.13 (75.7)	72.03 (72.0)	76.96 (76.5)	-
4-β-GlcA	103.20 (103.3)	73.78 (73.6)	74.28 (74.4)	-	-	-
F	103.20 (103.3)	73.78 (73.6)	74.28 (74.4)	-	-	-
3-β-Gal	104.49 (104.0)	71.36 (71.1)	-	68.83 (68.6)	-	61.18 (61.10)
G	104.49 (104.0)	71.36 (71.1)	-	68.83 (68.6)	-	61.18 (61.10)

Groups of animals	TL	Cells/mm³ MN	PMN
N	5200 ± 230	3640 ± 159	1560 ± 121
Cg	73470 ± 2530^a	10238 ± 990^a	63232 ± 2406^a
Indo _{5 mg/kg}	66727 ± 2938^a	11909 ± 340.2^a	54818 ± 3654^a
KEPS _{400 mg/kg}	62625± 3647^a	11305 ± 815.0^a	51320 ± 4241^a
KEPSC _{400 mg/kg}	65851 ± 4437^a	12880 ± 1734^a	52971 ± 4016^a

Groups of animals	TL	Cells/mm³ MN	PMN
N	6767 ± 129.5	4897 ± 493.1	1870 ± 103.9
Cg	73167 ± 4718^a	9347 ± 1158^a	63820 ± 584.5^a
Indo _{5 mg/kg}	71800 ± 5530^a	12078 ± 1606^a	59722 ± 6316^a
KEPS_{50 mg/kg}	23501 ± 1159^a,b	3361 ± 34.6^a,b	20140 ± 325.2^a,b
KEPS_{100 mg/kg}	11211 ± 896.5^a,b,c	2119 ± 502.8^a,b	9092 ± 37.91^a,b,c
KEPS_{200 mg/kg}	12083 ± 840.9^a,b,c	1235 ± 294.3^a,b	10848 ± 356.5^a,b,c
KEPS_{400 mg/kg}	7072 ± 897.6^a,b,c,d	2125 ± 446.7^a,b	4947 ± 328.9^a,b,c,d

[1] *Correspondence: R. A. C. Gonçalves. Departamento de Farmácia. Universidade Estadual de Maringá. Avenida Colombo 5790, Maringá, Paraná, Brasil. Fone: +55-44-30115967. Email: racgoncalves@uem.br

Brasil

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Exopolysaccharides from Klebsiella oxytoca: anti-inflammatory activity

Abstract