Masson Pine Extract

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Androgens can cause oxidative stress damage in dermal papilla cells and promote the expression of transforming growth factor (TGF)-β1, a negative regulator of hair growth, while antioxidants can significantly inhibit this process. The nuclear factor E2-related factor 2-antioxidant response element (Nrf2-ARE) pathway is a key signaling pathway for cellular resistance to oxidative stress. Studies have confirmed that the Nrf2-ARE pathway can protect hair follicles against oxidative stress damage. Masson pine needle extract (PMNE) is extracted from the pine leaves of Masson pine. It is a natural substance mainly composed of flavonoids and has various biological activities such as antioxidant, anti-inflammatory, and antibacterial.

HDPC were used as the research object. HDPC were treated with 0 (control group), 0.1, 0.2, 0.4, 0.8, and 1.0 mmol/L H2O2 to establish the optimal conditions of HDPC oxidative stress in vitro. HDPC were transfected with nuclear factor E2-related factor 2 (Nrf2) interference fragment siRNA1, siRNA2, siRNA3 or overexpression plasmid pCMV6-XL5-Nrf2, and the expressions of Nrf2 mRNA and protein were detected by real-time fluorescence quantitative PCR and Western blotting, respectively. The cell activity and apoptosis rate of each group after transfection were detected under H2O2 conditions. HDPCs were routinely cultured and divided into groups for treatment: control group: normal culture without other treatments; dihydrotestosterone group: added with 0.03 μg/ml dihydrotestosterone; proanthocyanidin group: added with 0.03 μg/ml dihydrotestosterone and 6.00 μg/ml proanthocyanidin B2; different concentrations of PMNE groups: added with 0.03 μg/ml dihydrotestosterone culture medium, and treated with 1, 5, 25, and 100 μg/ml PMNE respectively; cell viability and apoptosis rate, relative fluorescence intensity of intracellular reactive oxygen species (ROS) and malondialdehyde (MDA) content, expression levels of Nrf2, quinone oxidoreductase 1 (NQO1), heme oxygenase 1 (HO-1), Kelch-like ECH-associated protein 1 (Keap1), transforming growth factor (TGF)-β1, Sma and Mad-related protein 2/3 (Smad2/3), and p-Smad2/3 were detected in each group. One-way analysis of variance was used for comparison among multiple groups, and LSD-t test was used for comparison among two groups.

Figure 1 qPCR detection of Nrf2 mRNA expression after transfection of human dermal papilla cells with Nrf2-siRNA or overexpression plasmid 1A: 24 and 48 h after transfection, the expression of Nrf2 mRNA in Nrf2-siRNA1, Nrf2-siRNA2, and Nrf2-siRNA3 groups were all significant. significantly lower than the blank group and control group; 1B: 24 and 48 h after transfection, the expression of Nrf2 mRNA in the Nrf2 overexpression group was significantly higher than that of the blank group and control group.

When the HDPC cell viability was 75% ~ 85%, 0.4 mmol/L H2O2 was selected as the optimal treatment concentration for HDPC oxidative stress in vitro. The Nrf2 mRNA and protein expressions in the Nrf2-siRNA1, Nrf2-siRNA2, and Nrf2-siRNA3 groups were significantly lower than those in the blank group and the control group (all P < 0.05), and the cell apoptosis rates (12.50% ± 0.05%, 26.07% ± 0.05%, and 58.44% ± 1.03%) were significantly higher than those in the blank group (10.38% ± 0.64%) and the control group (13.05% ± 0.12%), all P < 0.05. The Nrf2 protein expression in the Nrf2-siRNA2 group was the lowest, and Nrf2-siRNA2 was selected as the best interference fragment for subsequent experiments. The expression of Nrf2 mRNA and protein in the Nrf2 overexpression group was significantly higher than that in the blank group and the control group (all P < 0.05), and the apoptosis rate was significantly lower than that in the blank group and the control group (all P < 0.05). Under the condition of 0.4 mmol/L H2O2 treatment, the apoptosis rate of the Nrf2 overexpression group was significantly lower than that in the overexpression empty group (t = 3.66, P < 0.001), and the cell activity was higher than that in the overexpression empty group (t = 40.40, P < 0.001); the apoptosis rate of the Nrf2-siRNA2 group was significantly higher than that in the control group (t = 13.13, P < 0.001), and the cell activity was significantly lower than that in the control group (t = 67.37, P < 0.001). In the PMNE treatment experiment, the cell activity of the proanthocyanidin group and different concentrations of PMNE groups were significantly higher than that of the dihydrotestosterone group (all P < 0.01), while the cell apoptosis rate was significantly lower than that of the dihydrotestosterone group (all P < 0.01); proanthocyanidin and different concentrations of PMNE could significantly inhibit the overexpression of ROS and MDA in HDPC cells induced by dihydrotestosterone (all P < 0.01); the protein expressions of Nrf2, NQO1, and HO-1 in the proanthocyanidin group and 5, 25, and 100 μg/ml PMNE groups were significantly higher than those in the dihydrotestosterone group (all P < 0.05), and the protein expressions of Keap1 and TGF-β1 and the phosphorylation levels of Smad2/3 in the proanthocyanidin group and 25 and 100 μg/ml PMNE groups were significantly lower than those in the dihydrotestosterone group (all P < 0.05).

Nrf2 plays an important role in resisting oxidative stress damage in HDPC. PMNE may have a significant protective effect on HDPC by activating the Nrf2 antioxidant response element pathway.

YTBIO mainly supplies Masson Pine Extract etc. The company was established in 2014 and has many years of R&D and production experience. We are committed to the research and development, production and sales of raw materials for the 21st century health industry. The certificates obtained so far include ISO9001, ISO22000, HALAL, KOSHER, HACCP, FDA,etc. We always maintain the highest requirements for product quality.

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