Protective Effect of Artocarpus heterophyllus Lam. (Jackfruit) Polysaccharides on Liver Injury Induced by Cyclophosphamide in Mice

Abstract

In recent years, Artocarpus heterophyllus Lam. (jackfruit) polysaccharides (namely JFP-Ps) have attracted much attention due to their multiple biological activities. This study aimed to explore the protective effects and the underlying mechanisms of JFP-Ps on cyclophosphamide (Cp)-induced liver damage. The protective effect of JFP-Ps was evaluated using HE staining, antioxidant testing, enzyme-linked immunosorbent assay (ELISA), real-time quantitative polymerase chain reaction (RT-qPCR), Western blot and ultra-performance liquid chromatography equipped with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) metabolomics analysis. The results showed that Cp caused pathological liver damage, activated oxidative stress and downregulated cytokine expression, while JFP-Ps treatment was found to exert antioxidant effects and play immune regulatory roles through mitogen-activated protein kinase/nuclear factor-κB (MAPK/NF-κB) related inflammation and cell apoptosis pathways to protect the Cp-induced liver injury. Metabolomic results showed that the liver-protective effects of JFP-Ps were mainly related to aminoacyl transfer ribonucleic acid (tRNA) biosynthesis, sphingolipid metabolism, purine metabolism and the citrate cycle. These results indicate that JFP-Ps have great potential application in alleviating liver injury.

Keywords: jackfruit polysaccharides; liver injury; metabolomics; protective effects.

Figure 1

Effects of JFP-Ps on liver histopathological characteristics (original magnification 40×, bar 20 μm). NC: normal control group; MC: model control group; LG: JFP-Ps low-dose group at 50 mg JFP-Ps/kg body weight; MG: JFP-Ps medium-dose group at 100 mg JFP-Ps/kg body weight; HG: JFP-Ps high-dose group at 200 mg JFP-Ps/kg body weight.

Figure 2

Effect of JFP-Ps on the mRNA expression of inflammation-related genes in the liver. Compared with NC group: * p < 0.05, ** p < 0.01; Compared with MC group: ^# p < 0.05, ^## p < 0.01. NC: normal control group; MC: model control group; LG: JFP-Ps low-dose group at 50 mg JFP-Ps/kg body weight; MG: JFP-Ps medium-dose group at 100 mg JFP-Ps/kg body weight; HG: JFP-Ps high-dose group at 200 mg JFP-Ps/kg body weight.

Figure 2

Effect of JFP-Ps on the mRNA expression of inflammation-related genes in the liver. Compared with NC group: * p < 0.05, ** p < 0.01; Compared with MC group: ^# p < 0.05, ^## p < 0.01. NC: normal control group; MC: model control group; LG: JFP-Ps low-dose group at 50 mg JFP-Ps/kg body weight; MG: JFP-Ps medium-dose group at 100 mg JFP-Ps/kg body weight; HG: JFP-Ps high-dose group at 200 mg JFP-Ps/kg body weight.

Figure 3

Effect of JFP-Ps on protien expression of the proteins involved in inflammation-related pathways. Compared with NC group: * p < 0.05, ** p < 0.01; Compared with MC group: ^# p < 0.05, ^## p < 0.01. NC: normal control group; MC: model control group; LG: JFP-Ps low-dose group at 50 mg JFP-Ps/kg body weight; MG: JFP-Ps medium-dose group at 100 mg JFP-Ps/kg body weight; HG: JFP-Ps high-dose group at 200 mg JFP-Ps/kg body weight.

Figure 3

Effect of JFP-Ps on protien expression of the proteins involved in inflammation-related pathways. Compared with NC group: * p < 0.05, ** p < 0.01; Compared with MC group: ^# p < 0.05, ^## p < 0.01. NC: normal control group; MC: model control group; LG: JFP-Ps low-dose group at 50 mg JFP-Ps/kg body weight; MG: JFP-Ps medium-dose group at 100 mg JFP-Ps/kg body weight; HG: JFP-Ps high-dose group at 200 mg JFP-Ps/kg body weight.

Figure 4

Total Ion Chromatogram (TIC) map and metabolites heatmap in positive (A,B) and negative (C,D) electrospray ionization (ESI) modes. NC: normal control group; MC: model control group; LG: JFP-Ps low-dose group at 50 mg JFP-Ps/kg body weight; MG: JFP-Ps medium-dose group at 100 mg JFP-Ps/kg body weight; HG: JFP-Ps high-dose group at 200 mg JFP-Ps/kg body weight.

Figure 4

Total Ion Chromatogram (TIC) map and metabolites heatmap in positive (A,B) and negative (C,D) electrospray ionization (ESI) modes. NC: normal control group; MC: model control group; LG: JFP-Ps low-dose group at 50 mg JFP-Ps/kg body weight; MG: JFP-Ps medium-dose group at 100 mg JFP-Ps/kg body weight; HG: JFP-Ps high-dose group at 200 mg JFP-Ps/kg body weight.

Figure 5

Multivariate statistical analysis ((A,B) PCA score map; (C,D) OPLS-DA score map; (E,F) 200 permutation test).

Figure 6

Volcano map ((A): NC vs. MC in ESI+, (B): NC vs. MC in ESI−, (C): MC vs. LG in ESI+, (D): MC vs. LG in ESI−, (E): MC vs. MG in ESI+, (F): MC vs. MG in ESI−, (G): MC vs. HG in ESI+, (H): MC vs. HG in ESI−) and Veen map ((I) ESI+; (J) ESI−). Red: Metabolites which pass both log2 Fold Change > 1.0 and p < 0.05 cut-offs and are up-regulated. Blue: Metabolites which pass both log2 Fold Change < −1.0 and p < 0.05 cut-offs and are down-regulated. Green: Metabolites which pass the p < 0.05 cut-off and fail to pass the |log2 Fold Change| > 1.0 cut-off. Grey: Metabolites which neither pass the p < 0.05 cut-off nor the |log2 Fold Change| > 1.0 cut-off.

Figure 6

Volcano map ((A): NC vs. MC in ESI+, (B): NC vs. MC in ESI−, (C): MC vs. LG in ESI+, (D): MC vs. LG in ESI−, (E): MC vs. MG in ESI+, (F): MC vs. MG in ESI−, (G): MC vs. HG in ESI+, (H): MC vs. HG in ESI−) and Veen map ((I) ESI+; (J) ESI−). Red: Metabolites which pass both log2 Fold Change > 1.0 and p < 0.05 cut-offs and are up-regulated. Blue: Metabolites which pass both log2 Fold Change < −1.0 and p < 0.05 cut-offs and are down-regulated. Green: Metabolites which pass the p < 0.05 cut-off and fail to pass the |log2 Fold Change| > 1.0 cut-off. Grey: Metabolites which neither pass the p < 0.05 cut-off nor the |log2 Fold Change| > 1.0 cut-off.

Figure 7

Pathway analysis. (A) metabolic pathway diagram; (B) Aminoacyl-tRNA biosynthesis; (C) Sphingolipid metabolism; (D) Purine metabolism; (E) Glutathione metabolism. The colors from yellow to red represent metabolites with different level of significance, red being more significant and yellow less significant respectively. The compounds highlighted in red font represent the metabolic markers in the metabolic pathways.