Anti-Hair Loss Potential of Perilla Seed Extracts: In Vitro Molecular Insights from Supercritical Fluid Extraction

Abstract

Perilla seed has long been recognized in traditional diets for its health-promoting properties, but its potential role in hair loss prevention remains underexplored. This study compared three extraction methods-maceration (MAC), screw pressing (SC), and supercritical fluid extraction (SFE)-to determine their efficiency in recovering bioactive compounds and their effects on androgenetic alopecia (AGA)-related pathways. The SFE extract contained the highest levels of polyunsaturated fatty acids and tocopherols, while MAC uniquely recovered a broader range of polyphenols. Among all extracts, SFE-derived perilla seed extract showed the most consistent biological effects, promoting proliferation of human hair follicle dermal papilla cells (HFDPCs) by 139.4 ± 1.1% at 72 h (p < 0.05). It also reduced TBARS and nitrite levels in HFDPCs to 66.75 ± 0.62% of control and 0.87 ± 0.01 μM, respectively, indicating strong antioxidant and anti-inflammatory effects. Importantly, the SFE extract significantly downregulated SRD5A1-3 and TGF-β1 expression-key genes involved in androgen-mediated hair follicle regression-outperforming finasteride, dutasteride, and minoxidil in vitro by approximately 1.10-fold, 1.25-fold, and 1.50-fold, respectively (p < 0.05). These findings suggest that perilla seed extract obtained via supercritical fluid extraction may offer potential as a natural candidate to prevent hair loss through multiple biological mechanisms. These in vitro results support its further investigation for potential application in functional food or nutraceutical development targeting scalp and hair health.

Keywords: 5α-reductase inhibition; Perilla frutescens (L.) Britt.; androgenetic alopecia; maceration; screw extraction; transforming growth factor-beta.

Figure 1

Proliferation of HFDPCs at 0, 12, 24, 48, 72 h, and 1 week upon exposure to perilla seed extracts or the standard treatment (minoxidil), at 0.031 mg/mL, relative to the untreated control. Data are reported as mean ± SD. Abbreviations: MAC-PS, macerated perilla seed extract; SFE-PS, supercritical fluid-extracted perilla seed extract; SC-PS, screw-pressed perilla seed extract; ND, not detected.

Figure 2

The inhibitory effect of perilla seed extracts on nitrite production is evaluated in lipopolysaccharide (LPS)-induced RAW 264.7 (A) cells and HFDPCs (B). Results are compared against a negative control (LPS-induced cells without pre-treatment), standard treatment (DF: diclofenac sodium), and untreated control at 0.125 mg/mL. Data are reported as mean ± SD for each sample. Statistical comparisons are applied through a one-way ANOVA followed by Tukey’s post hoc test. Distinct lowercase letters (a–f) represent statistically significant differences among groups in each assay (p < 0.05). Abbreviations: MAC-PS, macerated perilla seed extract; SFE-PS, supercritical fluid-extracted perilla seed extract; SC-PS, screw-pressed perilla seed extract.

Figure 3

The effect of perilla seed extracts on TBARS levels in hydrogen peroxide (H₂O₂)-induced HFDPCs evaluated in comparison with the negative control (H₂O₂-induced without pre-treatment), standard treatment (L-ascorbic acid), and untreated control, at 0.125 mg/mL. Data are reported as mean ± SD for each sample. Statistical comparisons are applied through a one-way ANOVA followed by Tukey’s post hoc test. Distinct lowercase letters (a–f) represent statistically significant differences among groups in each assay (p < 0.05). Abbreviations: MAC-PS, macerated perilla seed extract; SFE-PS, supercritical fluid-extracted perilla seed extract; SC-PS, screw-pressed perilla seed extract.

Figure 4

Gene expression related to the androgen pathway is analyzed following treatment with perilla seed extracts: (A) SRD5A1, (B) SRD5A2, and (C) SRD5A3 in DU-145 cells, and (D) SRD5A1, (E) SRD5A2, and (F) SRD5A3 in HFDPCs. These effects are compared to those of conventional therapeutic treatments (dutasteride, finasteride, and minoxidil) at 0.125 mg/mL. Data are reported as mean ± SD for each sample. Statistical comparisons are applied through a one-way ANOVA followed by Tukey’s post hoc test. Distinct lowercase letters (a–g) represent statistically significant differences among groups in each assay (p < 0.05). Abbreviations: MAC-PS, macerated perilla seed extract; SFE-PS, supercritical fluid-extracted perilla seed extract; SC-PS, screw-pressed perilla seed extract.

Figure 5

The expression of transforming growth factor-beta 1 (TGF-β1) gene is analyzed following treatment with perilla seed extracts in HFDPCs. These effects are compared with those of conventional therapeutic treatments (dutasteride, finasteride, and minoxidil) at a concentration of 0.125 mg/mL. Data are reported as mean ± SD for each sample. Statistical comparisons are applied through a one-way ANOVA followed by Tukey’s post hoc test. Distinct lowercase letters (a–e) represent statistically significant differences among groups in each assay (p < 0.05). Abbreviations: MAC-PS, macerated perilla seed extract; SFE-PS, supercritical fluid-extracted perilla seed extract; SC-PS, screw-pressed perilla seed extract.