Network Pharmacology and In Vivo Evaluation of Lycium barbarum Polysaccharide in Preventing Perfluorooctanoic Acid-Induced Damage in Broilers

Abstract

To examine the guarding effects of Lycium barbarum polysaccharides (LBP) on liver and gut injury induced by perfluorooctanoic acid (PFOA) in broilers. Initially, a dose selection experiment for PFOA modeling was conducted. A total of 160 one-day-old broilers were randomly assigned to four groups. The drinking water was supplemented with 0 mg/L PFOA (CON group), 0.5 mg/L PFOA (L-PFOA group), 1 mg/L PFOA (M-PFOA group), and 1.5 mg/L PFOA (H-PFOA group). The results indicated that compared to the CON group, the H-PFOA group showed significant increases liver function indicators, while antioxidant enzyme levels were significantly downregulated. Additionally, the H-PFOA group exhibited significant damage to the liver and intestinal tissue morphology. Subsequently, network pharmacology methods were employed to identify target protein information for LBP in preventing PFOA-induced damage. Protein-protein interaction analysis, GO functional annotation, and KEGG pathway enrichment analysis were performed. The network pharmacology results revealed a total of 100 key protein targets, with core targets mainly enriched in the AGE-RAGE and FOXO signaling pathways. Finally, animal experiments were conducted to verify. A total of 240 one-day-old broilers were randomly divided into six groups: CON group as blank control, PFOA group supplemented with 1.5 mg/L PFOA, l-LBP, M-LBP, and H-LBP groups further supplemented with 0.4 %, 0.6 %, and 0.8 % LBP respectively on a base of 1.5 mg/L PFOA, with LBP group as a control supplemented only with 0.8 % LBP in water. The results demonstrated that versus the CON group, the H-LBP group dropped considerably in daily weight gain, liver index, and pro-inflammatory cytokine levels, while antioxidant enzyme levels and intestinal tight junction protein mRNA expression were significantly upregulated. Moreover, the H-LBP group showed significant improvement in liver and intestinal tissue morphology. qPCR results revealed that PFOA exposure activated the AKT/FOXO1 signaling pathway, which was inhibited by LBP. In conclusion, LBP prevent PFOA-induced damage via a multi-component, multi-target, and multi-pathway mechanism.

Keywords: AKT/FOXO1 Pathway; Broiler; Lycium barbarum polysaccharide; Network pharmacology; Perfluorooctanoic acid.

Graphical abstract

Fig 1

Analysis of PFOA on liver and intestinal related indicators in broiler chickens. (a) liver antioxidant indicators; (b)intestinal antioxidant indicators. CON is control group; l-PFOA is 0.5 mg/L PFOA; M-PFOA is1mg/L PFOA; H-PFOA is 1.5 mg/L PFOA. Results are shown as mean ± SD (n = 8). Statistical evaluation was done using one-way ANOVA. Versus the CON group, *P < 0.05, **P < 0.01.

Fig 2

Screening of potential targets for LBP against PFOA. (a) Venn plot; (b) LBP-monosaccharide-target-disease network diagram.

Fig 3

PPI network diagram of LBP and PFOA intersection targets. (a) PPI network diagram; (b) PPI network diagram sorted by degree value; (c) Top 10 core targets; (d-f) Three gene clusters of MCODE module. The color depth, circle size, and target degree value correspond to each other.

Fig 4

GO functional annotation and KEGG signaling pathway enrichment. (a) GO functional annotation; (b) KEGG signaling pathway enrichment; (c) Module 1 KEGG signaling pathway enrichment Top 10 core targets.

Fig 5

LBP′s effect on pathological tissue slices and oxidative stress induced by PFOA. (a) Liver histopathology, → means vacuolar denaturation, ▲ means nuclear shrinkage; (b) Intestinal histopathology, → means villus breakage, ▲ means the villi have fallen of; (c) liver antioxidant indicators; (d)intestinal antioxidant indicators.CON is control group; PFOA is 1.5 mg/L PFOA model group; l-LBP is 1.5 mg/L PFOA+0.4 % LBP; M-LBP is 1.5 mg/L PFOA+0.6 % LBP; H-LBP is 1.5 mg/L PFOA+0.8 % LBP; LBP is 0.8 % LBP control group. . Results are shown as mean ± SD (n = 8). Statistical evaluation was done using one-way ANOVA. Versus the CON group, *P < 0.05, **P < 0.01; Versus the PFOA group, #P < 0.05, ##P < 0.01.

Fig 6

Effects of LBP on liver and intestinal related indicators. (a) liver pro-inflammatory factor levels; (b) Intestinal pro-inflammatory factor levels; (c) effect of LBP on mRNA expression levels of intestinal tight junction protein; (d) correlation analysis of the impact of LBP intervention on liver and intestinal indicators. CON is control group; PFOA is 1.5 mg/L PFOA model group; l-LBP is 1.5 mg/L PFOA+0.4 % LBP; M-LBP is 1.5 mg/L PFOA+0.6 % LBP; H-LBP is 1.5 mg/L PFOA+0.8 % LBP; LBP is 0.8 % LBP control group. Results are shown as mean ± SD (n = 8). Statistical evaluation was done using one-way ANOVA. Versus the CON group, *P < 0.05, **P < 0.01; Versus the PFOA group, #P < 0.05, ##P < 0.01.

Fig 7

The effect of LBP intervention on liver and intestinal AKT/FoxO1 signaling pathway signaling pathways. (a) liver; (b) intestinal; (c) intervention mechanism of LBP on PFOA model. CON is control group; PFOA is 1.5 mg/L PFOA model group; l-LBP is 1.5 mg/L PFOA+0.4 % LBP; M-LBP is 1.5 mg/L PFOA+0.6 % LBP; H-LBP is 1.5 mg/L PFOA+0.8 % LBP; LBP is 0.8 % LBP control group. Results are shown as mean ± SD (n = 8). Statistical evaluation was done using one-way ANOVA. Versus the CON group, *P < 0.05, **P < 0.01; Versus the PFOA group, #P < 0.05, ##P < 0.01.