Antithrombotic effect and plasma pharmacochemistry of Justicia Procumbens L

Abstract

Importance: Justicia procumbens L. contains lignans with remarkable antiplatelet aggregation effects. Its ethyl acetate extract has demonstrated the efficacy of inhibiting platelet aggregation in preliminary tests. Identifying key components, elucidating the antithrombotic mechanism, and evaluating its potential for treating thrombotic diseases are of great significance.

Methods: Chemical analysis was conducted on the ethyl acetate extract of J. procumbens. Animal models of carotid artery and venous thrombosis, combined with proteomics analysis, were employed to study the antithrombotic mechanism.

Results: Chemical analysis identified justicidin B (JB) and chinensinaphthol methyl ether (CME) as the major active antithrombotic constituents. The extract inhibited platelet aggregation by regulating relevant signaling pathways, showing potential in preventing both venous and arterial thrombosis.

Conclusions: This study confirmed that the ethyl acetate extract of J. procumbens, especially the lignan components JB and CME, can effectively inhibit platelet aggregation, showing promising prospects in treating thrombotic diseases. This provides ideas for the development of health products based on J. procumbens and lays a foundation for further clinical exploration.

Fig 1. (A) Disease and compound Venn diagram. (B) PPI network of intersection targets. (C) GO functional enrichment analysis of intersection targets. (D) KEGG enrichment analysis of intersection targets. (E) D-C-T-P network diagram of intersection targets. The abbreviations that appear in the pictures are presented in S3, S4, and S5 of the Supporting Information.

Fig 2. UPLC-MS of the chemical composition and plasma components of ethyl acetate extract from J. procumbens.

(A) Methanol. (B) Ethyl acetate extract. (C) Blank plasma. (D) Ethyl acetate extract components in plasma.

Fig 3. (A) Effective extract chromatogram. (a) JB, (b) CME, and (c) NJB. (B, C, and D) Key compound structures in J. procumbens effective extract. (B) Justicidin B., (C) chinensinaphthol methyl ether, and (D) neojusticin B.

Fig 4. Clot retraction.

Note: the black line represents Aspirin, which inhibits clot retraction; the brown line stands for The ethyl acetate extract, showing a weak promotion of clot retraction; the red line indicates The effective extract, strongly promoting clot retraction; the blue line denotes Justicidin B, promoting clot retraction in the early stage but slowing down later; and the green line represents Chinensinaphtholmethyl ether (CME), significantly inhibiting clot retraction.

Fig 5. (A) Vascular status during surgery. (B) Weight of thrombi in each group. (C) Thrombosis images from animal experiments in different groups with deep vein thrombosis. (D) Weight of thrombi in each group.

Fig 6. H&E staining results.

Fig 7. (A) GO analysis of the down-regulated effective extract proteins in proteomics. (B) GO analysis of the up-regulated effective extract proteins in proteomics. (C) KEGG analysis of the down-regulated effective extract proteins in proteomics. (D) KEGG analysis of the up-regulated effective extract proteins in proteomics. (E) PPI diagram of the down-regulated proteins in proteomics. Note: (C) and (D): The abscissa represents the pathway name, and the ordinate represents the enrichment ratio [which is the ratio of the number of proteins enriched in this pathway (protein number) to the number of proteins annotated to the pathway (background number). The larger the ratio, the greater the degree of enrichment]. The color gradient of the columns represents the significance of enrichment. By default, the darker the color, the more significantly enriched the KEGG term. Among them, those with a P or FDR value < 0.001 are marked as ***, those with P or FDR values < 0.01 are marked as **, and those with P or FDR values < 0.05 are marked as *.

Fig 8. Molecular docking experiment results.

(A) CME and F9. (B) JB and F9. (C) CME and F12. (D) JB and F12. (E) CME and F2rl2. (F) JB and F2rl2. (G) CME and Prkce. (H) JB and Prkce. (I) CME and C3. (J) JB and C3. (K) CME and C1q. (L) JB and C1q. (M) CME and C8β. (N) JB and C8β. (O) CME and C7. (P) JB and C7.The red color in the small molecule represents oxygen atoms, and the other color parts such as rose, blue, and yellow represent carbon atoms in general.