A Mixture of Lactobacillus fermentum HFY06 and Arabinoxylan Ameliorates Dextran Sulfate Sodium-Induced Acute Ulcerative Colitis in Mice

Figures & data

Figure 1 Animal experiment design.

Table 1 Evaluation of the Disease Activity Index (DAI)

Stool Consistency	Occult/Gross Bleeding	Weight Loss (%)	Score
Normal	Normal (-)	0	0
Soft but shaped	Occult blood positive (+)	1–5	1
Semi loose stool	Occult blood (++)	>5–10	2
Loose stool	Gross bleeding (+++)	>10–15	3
Heavier loose stools	Gross bleeding(>+++)	>15	4

Note: DAI = (weight loss score+fecal consistency score+occult/gross bleeding)/3.

Table 2 Primer Sequence List

Gene Name	Sequence
NF-κBp65	Forward: 5ʹ-ATGGCAGACGATGATCCCTAC-3’
	Reverse: 5ʹ-CGGAATCGAAATCCCCTCTGTT-3’
IκB-α	Forward: 5′-TGAAGGACGAGGAGTACGAGC-3′
	Reverse: 5′-TGCAGGAACGAGTCTCCGT-3′
iNOS	Forward: 5′-GTTCTCAGCCCAACAATACAAGA-3′
	Reverse: 5ʹ-GTGGACGGGTCGATGTCAC-3′-3’
COX-2	Forward: 5ʹ-GGTGCCTGGTCTGATGATG-3’
	Reverse: 5ʹ-TGCTGGTTTGGAATAGTTGCT-3’
TNF-α	Forward: 5ʹ-CAGGCGGTGCCTATGTCTC-3’
	Reverse: 5ʹ-GCTGCAACAGGGGGTAACAT-3’
β-actin	Forward: 5ʹ-CATGTACGTTGCTATCCAGGC-3’
	Reverse: 5ʹ-CTCCTTAATGTCACGCACGAT-3’

Table 3 Comparison of Anti-Inflammatory Effects Between Control Strain and L. fermentum HFY06 Strain

Group	IL-6 (ng/L)	IL-10 (ng/L)	TNF-α (ng/L)	IFN-γ (ng/L)
LB	54.20±3.17	230.16±9.36	424.71±12.82	90.42±6.99
HFY06	50.91±2.36	253.41±8.79***	334.21±55.45**	73.58±8.13*

Notes: The data are expressed as the mean ± SD. LB: 2.5% DSS with Lactobacillus delbrueckii subsp. Bulgaricus. HFY06: 2.5% DSS with L. fermentum HFY06 (1.0 × 10¹⁰ CFU). *P < 0.05, **P < 0.01, ***P < 0.001 compared with the LB group.

Figure 2 Effect of AX combine L. fermentum HFY06 supplementation on (A) body weight and (B) DAI of mice.

Table 4 Serum Levels of SOD, NO and MDA in Mice

Group	SOD (U/mL)	NO (µmol/L)	MDA (nmol/mL)
Normal	55.69 ± 2.66^c	6.18 ± 0.62^a	1.51 ± 0.17^a
Model	34.72 ± 2.60^a	10.5 ± 0.32^d	5.65 ± 0.48^d
HFY06	36.61 ± 4.75^a	8.08 ± 0.41^c	2.57 ± 0.25^b
AX	39.24 ± 4.41^ab	8.45 ± 0.77^c	3.30 ± 0.24^c
AX+HFY06	44.52 ± 6.46^b	7.19 ± 0.18^b	1.47 ± 0.11^a

Notes: The data are expressed as the mean ± SD. ^a–d Values in the same column with different letter superscripts indicate significant difference (p < 0.05). HFY06: 2.5% DSS with L. fermentum HFY06 (1.0 × 10¹⁰ CFU), AX: 2.5% DSS with AX (200 mg/kg), AX+HFY06: 2.5% DSS with AX (200 mg/kg) and L. fermentum HFY06 (1.0 × 10¹⁰ CFU).

Table 5 Serum Levels of IL-1β, IL-6, IL-12, TNF-α, IFN-γ, and IL-10 in Mice

Group	IL-1β (ng/L)	IL-6 (ng/L)	IL-12 (ng/L)	TNF-α (ng/L)	IFN-γ (ng/L)	IL-10 (ng/L)
Normal	43.92±3.84^a	43.68±6.82^a	60.57±6.18^a	237.57±34.37^a	56.45±14.52^a	374.07±40.38^c
Model	66.98±8.30^c	64.10±5.96^c	72.32±5.06^b	536.47±34.92^c	91.92±6.62^c	294.78±24.36^a
HFY06	51.49±7.61^ab	50.91±2.36^ab	58.20±3.56^a	334.21±55.45^b	73.58±8.13^b	253.41±8.79^b
AX	55.27±5.81^b	55.11±9.00^b	58.01±8.67^a	541.09±16.26^c	69.72±14.18^ab	297.08±34.41^b
AX+HFY06	48.64±5.67^ab	45.39±5.57^a	55.58±4.05^a	307.22±29.85^b	63.51±7.19^ab	264.41±18.43^ab

Notes: The data are expressed as the mean ± SD. ^a–cValues in the same column with different letter superscripts indicate significant difference (p < 0.05). HFY06: 2.5% DSS with L. fermentum HFY06 (1.0 × 10¹⁰ CFU), AX: 2.5% DSS with AX (200 mg/kg), AX+HFY06: 2.5% DSS with AX (200 mg/kg) and L. fermentum HFY06 (1.0 × 10¹⁰ CFU).

Figure 3 Effect of AX combine L. fermentum HFY06 supplementation on myeloperoxidase (MPO) enzyme activity in colon tissue. HFY06: 2.5% DSS with L. fermentum HFY06 (1.0 × 10¹⁰ CFU), AX: 2.5%DSS with AX (200 mg/kg), AX+HFY06: 2.5% DSS with AX (200 mg/kg) and L. fermentum HFY06 (1.0 × 10¹⁰ CFU). Different letters above the histogram indicate significant differences (P<0.05).

Figure 4 Effect of AX combine L. fermentum HFY06 supplementation on H&E pathological of colon tissue in mice ((A) Normal, (B) Model, (C) L. fermentum HFY06, (D) AX, (E) L. fermentum HFY06 combined with AX). HFY06: 2.5% DSS with L. fermentum HFY06 (1.0 × 10¹⁰ CFU), AX: 2.5%DSS with AX (200 mg/kg), AX+HFY06: 2.5% DSS with AX (200 mg/kg) and L. fermentum HFY06 (1.0 × 10¹⁰ CFU). Arrow 1 indicates inflammatory cell infiltration, arrow 2 indicates a reduction in goblet cells, and arrow 3 indicates colonic mucosal erosion.

Figure 5 Effect of AX combine L. fermentum HFY06 supplementation on mRNA expression levels in colon tissues ((A) NF-κBp65, (B) IκB-α, (C) iNOS (D) COX-2, (E) TNF-α). HFY06: 2.5% DSS with L. fermentum HFY06 (1.0 × 10¹⁰ CFU), AX: 2.5%DSS with AX (200 mg/kg), AX+HFY06: 2.5% DSS with AX (200 mg/kg) and L. fermentum HFY06 (1.0 × 10¹⁰ CFU). Different letters above the histogram indicate significant differences (P<0.05).