Lactobacillus paracasei HY7015 and Lycopus lucidus Turcz. Extract Promotes Human Dermal Papilla Cell Cytoprotective Effect and Hair Regrowth Rate in C57BL/6 Mice

Abstract

Hair loss is a disease that requires accurate diagnosis and type-specific medical treatment. Many hair loss treatments have some side effects, such as hormone-related effects, so there is a need for safe and effective hair loss treatment. In this study, we investigated the effects of Lactobacillus paracasei HY7015 (HY7015) and Lycopus lucidus Turcz. (LT) extract on hair regrowth and protection. In vitro experiments were conducted to assess the effects of HY7015 and/or LT extract on human follicle dermal papilla cells (HFDPC) of cytoprotective functions such as proliferations, antioxidants, anti-inflammatory, and growth factor expressions. In animal experiments, we investigated hair regrowth rate, hair follicle formation and secretion of growth factors in telogenic C57BL/6 mice. We confirmed the cytoprotective effects of HY7015 and LT through regulations of proliferation, SOD and IL-1β in HFDPC. In mouse experiments, oral administration of HY7015 and LT promoted hair regrowth as well as hair follicle maturation in the dermal skin of C57BL/6 mice, and upregulated VEGF and IGF-1 growth factor levels in mouse serum. In summary, our data demonstrate that ingestions of HY7015 and LT can promote hair regrowth by enhancing cytoprotective effects and expressions of growth factors.

Keywords: Lactobacillus paracasei; Lycopus lucidus Turcz.; cytoprotective effects; hair regrowth.

Figure 1

The plant studied, Lycopus lucidus Turcz.

Figure 2

Cytoprotective effects of Lactobacillus paracasei HY7015 (HY7015) and/or Lycopus lucidus Turcz. (LT) extract in HFDPC. Effect of Lactobacillus paracasei HY7015 (HY7015) and/or Lycopus lucidus Turcz. (LT) extract on HFDPC proliferation (A). HFDPC were untreated (Control), or treated with medicinal yeast extract (MYE, 50 µg/mL), or with HY7015 (1 × 10⁶ CFU/mL) and/or LT (50 µg/mL) for 24 h. SOD1 activity (B) and IL-1β (C) were measured by ELISA kit. Data are presented as mean ± SE. Significant differences are indicated by * p < 0.05 and *** p < 0.001, relative to the control group. # p < 0.05 compared with H₂O₂ group. SE, standard error; MYE, medicinal yeast extract; HY7015, Lactobacillus paracasei HY7015; LT, Lycopus lucidus Turcz.; HY7015+LT, Lactobacillus paracasei HY7015 with Lycopus lucidus Turcz.

Figure 3

Effect of Lactobacillus paracasei HY7015 (HY7015) and/or Lycopus lucidus Turcz. (LT) extract on VEGF proteins (A) and mRNA (B) of HFPDC, VEGF proteins (C), and Vegf mRNA (D) of NIH3T3 fibroblasts. Protein levels of VEGF were measured by ELISA kit. Relative mRNA levels of VEGF were monitored by qPCR and normalized against GAPDH. Data are presented as mean ± SE. Significant differences are indicated by * p < 0.05 and *** p < 0.001, relative to the control group. SE, standard error; VEGF, vascular endothelial growth factor; MYE, medicinal yeast extract; HY7015, Lactobacillus paracasei HY7015; LT, Lycopus lucidus Turcz.; HY7015+LT, Lactobacillus paracasei HY7015 with Lycopus lucidus Turcz.

Figure 4

Effect of Lactobacillus paracasei HY7015 (HY7015) and/or Lycopus lucidus Turcz. (LT) extract on promoting hair regrowth in a telogenic mouse model. Shaved 7-week-old mice were fed a normal control diet (Control), or medicinal yeast extract (MYE, 200 mg/kg), or HY7015 (1 × 10⁸ CFU/day) and/or LT extract (100 mg/kg) daily for 5 weeks. (A) Photographs of mouse dorsal skin by groups at week 5. (B) Hair regrowth rates during the 5 weeks period. (C) Weekly assessment of hair regrowth rates in each group during the 5 weeks period. Data are presented as mean ± SE. Significant differences are indicated by *** p < 0.001, relative to the control group. SE, standard error; MYE, medicinal yeast extract; HY7015, Lactobacillus paracasei HY7015; LT, Lycopus lucidus Turcz.; HY7015+LT, Lactobacillus paracasei HY7015 with Lycopus lucidus Turcz.

Figure 5

Oral administration of Lactobacillus paracasei HY7015 (HY7015) and/or Lycopus lucidus Turcz. (LT) extract promotes formation of hair follicles in telogenic mice. H&E staining histology analysis of (A) transverse sections. Slides were prepared by cutting the skin tissue perpendicular to the tail to observe mouse hair follicles. (B) Longitudinal sections for dermal layer thickness measurements of mouse dorsal skin tissues. Arrows indicate the presence of hair follicles in skin. (C) Numbers of hair follicles in dorsal skin layer samples (×10⁴/µm²). (D) Dermal skin thickness (μm). Data are presented as mean ± SE. Significant differences are indicated by *** p < 0.001, relative to the control group. SE, standard error; MYE, medicinal yeast extract; HY7015, Lactobacillus paracasei HY7015; LT, Lycopus lucidus Turcz.; HY7015+LT, Lactobacillus paracasei HY7015 with Lycopus lucidus Turcz.

Figure 6

Effect of Lactobacillus paracasei HY7015 (HY7015) and/or Lycopus lucidus Turcz. (LT) extract on growth factor secretion in mouse serum. (A) VEGF levels in mouse serum. (B) IGF-1 levels in mouse serum. VEGF and IGF-1 secretions were measured by ELISA. Data are presented as mean ± SE. Significant differences are indicated by * p < 0.05, ** p < 0.01, and *** p < 0.001, relative to the control group. SE, standard error; MYE, medicinal yeast extract; HY7015, Lactobacillus paracasei HY7015; LT, Lycopus lucidus Turcz.; HY7015+LT, Lactobacillus paracasei HY7015 with Lycopus lucidus Turcz.; ELISA, enzyme-linked immunosorbent assay.

Figure 7

Effect of rosmarinic acid on VEGF mRNA expressions in HFDPC. Relative mRNA levels of VEGF were monitored by qPCR and normalized against GAPDH. Data are presented as mean ± SE. Significant differences are indicated by * p < 0.05 and *** p < 0.001, relative to the control group. SE, standard error; VEGF, vascular endothelial growth factor; MYE, medicinal yeast extract; RA, rosmarinic acid.

Figure 8

The flowchart of in vitro and in vivo.