Asparagus cochinchinensis Extract Alleviates Metal Ion-Induced Gut Injury in Drosophila: An In Silico Analysis of Potential Active Constituents

Abstract

Metal ions and sulfate are components of atmospheric pollutants that have diverse ways of entering the human body. We used Drosophila as a model to investigate the effect of Asparagus cochinchinensis (A. cochinchinensis) extracts on the gut and characterized gut homeostasis following the ingestion of metal ions (copper, zinc, and aluminum). In this study, we found that the aqueous A. cochinchinensis extract increased the survival rate, decreased epithelial cell death, and attenuated metal ion-induced gut morphological changes in flies following chronic exposure to metal ions. In addition, we screened out, by network pharmacology, six natural products (NPs) that could serve as putative active components of A. cochinchinensis that prevented gut injury. Altogether, the results of our study provide evidence that A. cochinchinensis might be an effective phytomedicine for the treatment of metal ion-induced gut injury.

Figure 1

A. cochinchinensis improves the survival rate of flies that have ingested metal ions. (a) The survival rates of AC extract and control flies fed with 7 mM CuSO₄. (b) The survival rates of AC extract and control flies fed with 7 mM ZnSO₄. (c) The survival rates of AC extract and control flies fed with 20 mM KAl(SO₄)₂·12H₂O. Three replicates were used for the determination of survival rates. P value was calculated by the log-rank test. ^∗∗∗ P < 0.001 was considered statistically significant.

Figure 2

A. cochinchinensis protects gut epithelial cells from metal ion-induced cell death. (a) The effect of AC extract on Cu-induced epithelial cell death. (b) The effect of AC extract on Zn-induced epithelial cell death. (c) The effect of AC extract on Al-induced epithelial cell death. ((d)–(f)) The comparison of gut necrotic cell quantity between control and AC group. The number of necrotic cells was quantified using ImageJ software (10–15 guts were examined to quantify necrotic cells for each group). P value was calculated by Student's t-test. ^∗∗∗ P < 0.001; ^∗∗ P < 0.01. Con: control, flies fed with metal ions without AC for 96 h; AC: flies fed with metal ions with A. cochinchinensis (AC) for 96 h. 7-AAD: 7-amino-actinomycin D, dead cells (red signal); DAPI: 4′,6-diamidino-2-phenylindole, nucleus (blue signal); 7-AAD/DAPI (red and blue signal). Scale bar: 100 μm.

Figure 3

A. cochinchinensis prevents metal ion-induced gut atrophy. Con: control, flies fed with metal ion without AC for 96 h; AC: A. cochinchinensis, flies fed with metal ion with AC for 96 h. All experiments were independently performed three times. ^∗∗∗ P < 0.001; ns: no significant difference.

Figure 4

The NP-target network. The target protein was expressed by a node, and edges represented the relationship between NPs and the target protein. 11-DH: corticosteroid 11-beta-dehydrogenase isozyme 1; GR: glucocorticoid receptor; PPAR-gamma: peroxisome proliferator-activated receptor gamma. UNPD ID: Universal Natural Products Database identification.

Figure 5

Distribution of the six molecular descriptors of AC-NPs and FDA-approved drugs. H Acceptors: number of hydrogen bond receptors; H Donors: number of hydrogen bond donors; Rotatable Bonds: number of rotatable bonds; Rings: number of rings. Approved drugs: FDA-approved drugs; AC-NPs: A. cochinchinensis natural products.