A New Strategy for the Rapid Identification and Validation of the Direct Targets of Aconitine-Induced Cardiotoxicity

Figures & data

Figure 1 Venn diagram of aconitine and cardiotoxicity-related targets (A); PPI network diagram (B); GO enrichment analysis: BP, Biological process; CC, cellular component; MF, molecular function. (C); Top 20 KEGG signalling pathways bubble chart (D); “Aconitine–Targets–Pathways” network diagram (E).

Figure 2 Effects of different concentrations of aconitine on H9c2 cell viability (A). All data are shown as mean ± SD (n = 6). **p <0.01 versus the normal group; SDS-PAGE results of protein digested with 1:100 pronase: protein ratio after incubation with aconitine (B–D); (B–D) represent three repetitions of the experiments (n = 3).

Table 1 The Information of Direct Target Proteins Fished by DARTS Technology

Accession	Description	Gene	MW [kDa]
D3ZU40	Transmembrane protein 104	GN=Tmem104	55.6
D3ZNJ9	Tripartite motif-containing 7	GN=Trim7	56.9
A0A140UHX9	CCZ1 homolog B	GN=Ccz1b	55.6
D4AAD2	Serine protease 53	GN=Prss53	57.5
Q5XI07	Lipoma-preferred partner homolog	GN=Lpp	68.2
D3ZWA5	Kinesin-associated protein 3	GN=Kifap3	84.2
P50393	Cytosolic phospholipase A2	GN=cPla2	85.7
B4F7A2	Armc8 protein (Fragment)	GN=Armc8	75.3
Q5M7U2	Origin recognition complex, subunit 5	GN=Orc5	50.1
A0A0G2JYF7	Catenin alpha 1	GN=Ctnna1	89.3
P0C6P7	Protein fem-1 homolog B	GN=Fem1b	70.2
A1L1K3	Anaphase-promoting complex subunit 5	GN=Anapc5	81.7
Q6EHI2	Butyrophilin (Fragment)	GN=Btn1a1	57.0
D3ZAB1	Lactotransferrin	GN=Ltf	79.8
A2RRU1	Glycogen [starch] synthase, muscle	GN=Gys1	84.0

Figure 3 PPI network diagram of 152 core targets from network pharmacology and 15 direct targets from DARTS.

Figure 4 Molecular docking models of aconitine with 4 direct target proteins in 3D and 2D diagram. Aconitine with (A) cPLA2 (PDB: 1CJY); (B) GYS1 (PDB: 3CX4); (C) CTNNA1 (PDB: 4EHP); (D) ORC5 (PDB: 5UI7).

Figure 5 Potential toxicity mechanism of aconitine by acting on direct target cPLA2.

Figure 6 AFM analysis. AFM images (2D and 3D) of (A) cPLA2 and (B) aconitine acting on the protein.