# Network Pharmacology of the Phytochemical Content of Sunflower Seed (Helianthus annuus L.) Extract from LC-MS on Wound-Healing Activity and the In Vitro Wound Scratch Assay

## TL;DR

Sunflower seed extract promotes wound healing by enhancing cell migration and has antioxidant properties, with chlorogenic acid being a key compound.

## Contribution

This study identifies bioactive compounds in sunflower seed extract and their mechanisms in wound healing using network pharmacology and molecular docking.

## Key findings

- Sunflower seed extract at 10 µg/mL achieved complete wound closure in 24 hours in HaCaT cells.
- Chlorogenic acid showed the highest binding affinity to wound-healing targets like NF-κB1, EGFR, and MMP9.
- The extract exhibited moderate-to-strong antioxidant activity and favorable pharmacokinetic properties.

## Abstract

Sunflower seeds have been reported to be a healthy natural source of polyphenols. This study aimed to explore the mechanisms of potential compounds in sunflower seed extract involved in wound healing; major compounds were investigated through network pharmacology and molecular docking. In an in vitro wound-healing assay applied using an immortalised human keratinocyte (HaCaT) cell model, 10 µg/mL of the sunflower seed extract promoted cell migration in HaCaT cells and led to complete wound closure after 24 h; at a 1 µg/mL concentration, it led to complete wound closure after 72 h. The sunflower seed extract presented moderate-to-strong antioxidant activity. Liquid chromatography–mass spectrometry and high-performance liquid chromatography were used to identify the major compounds present in the sunflower seed extract. Forty-seven compounds were identified, among which chlorogenic acid was the most abundant phenolic compound. Network pharmacology was used to identify wound-healing-related targets. In total, 252 proteins were linked to the 47 compounds. Cyto-Hubba analysis identified 10 hub proteins with a strong correlation with wound healing. Molecular docking was used to assess the ability of the major compounds in the sunflower seed extract to combat NF-κB1, EGFR, and MMP9. Chlorogenic acid showed higher binding affinity to all targets. Moreover, its pharmacokinetic properties were well distributed in the plasma (VDss = 0.377 log L/kg), and they were not a carcinogen and did not cause skin sensitisation. In conclusion, the findings suggest that the sunflower seed extract is a potential source of bioactive compounds that can enhance wound healing and can be developed to create a transdermal application.