Phenolics isolated from Parnassia palustris reduces melanogenesis in mouse B16F10 melanoma cells : 마우스 유래 B16F10 흑색종 세포에서 Parnassia palustris로부터 추출한 페놀성 물질의 멜라닌 생합성 억제 효능

다국어 초록 (Multilingual Abstract)

Melanin protects human skin from UV damage, but its excessive production causes hyperpigmentation and increases melanoma risk, making the regulation of melanogenesis a critical therapeutic target. Parnassia palustris is a traditional medicinal herb used for its antipyretic and anti-inflammatory properties. In the current study, we examines the in vitro antioxidant capacity and anti-melanogenic potential of P. palustris extracts obtained with distilled water (PPW) and 60% ethanol (PPE). DPPH and ABTS radical scavenging activities, antioxidant protection factor, and TBARS inhibition effects were conducted to analyze the antioxidant capacity, with PPE demonstrating effects comparable to or exceeding those of BHT. The potential to inhibit melanin production was analyzed through B16F10 melanoma cells, demonstrating that PPE markedly decreased intracellular melanin contents and suppressed cellular tyrosinase activity. Furthermore, treatment with PPE resulted in reduced protein levels of MITF, TYR, TRP- 1, and TRP-2, which are related to melanogenesis. Notably, downregulation of mRNA, including MC1R, Myo5a, and Rab27a was also observed, suggesting inhibition of both early melanogenic signaling and melanosome transport pathways. Collectively, these results indicate that PPE possesses properties that may help regulate skin hyperpigmentation. These results further suggest that PPE may serve as a valuable plant-derived resource for developing cosmeceutical products.

목차 (Table of Contents)

• Abstract 1
• 1. Introduction 2
• 2. Materials and methods 5
• 2.1. Reagents and instruments 5
• 2.2. Preparation of Parnassia palustris extracts 5
• 2.3. Phytochemical analysis 6
• 2.3.1. Quantification of the total phenolic content (TPC) 6
• 2.3.2. Quantification of the total flavonoid content (TFC) 7
• 2.4. Antioxidant assay 8
• 2.4.1. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay 8
• 2.4.2. 2,2′-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) cation radical decolorization assay 8
• 2.4.3. Antioxidant protection factor (PF) assay 9
• 2.4.4. Thiobarbituric acid reactive substances (TBARS) inhibition assay 10
• 2.5. Mushroom tyrosinase inhibition assay 11
• 2.6. Anti-melanogenesis properties of Parnassia palustris extracts in mouse B16F10 melanoma cells 12
• 2.6.1. Cell culture 12
• 2.6.2. Cell viability 13
• 2.6.3. Quantification of melanin content 14
• 2.6.4. Cellular tyrosinase inhibition assay 15
• 2.6.5. Western blot analysis 16
• 2.6.6. Real-time polymerase chain reaction (RT-qPCR) analysis 18
• 2.7. Statistical analysis 20
• 3. Results and Discussion 21
• 3.1. Total phenolic and flavonoid content in Parnassia palustris extracts 21
• 3.2. Antioxidant activity of Parnassia palustris extracts 23
• 3.2.1. DPPH radical scavenging activity 23
• 3.2.2. ABTS cation radical decolorization activity 24
• 3.2.3. Antioxidant protection factor (PF) activity 25
• 3.2.4. TBARS inhibition activity 26
• 3.3. Mushroom tyrosinase inhibition activity 28
• 3.4. Anti-melanogenesis effect of Parnassia palustris 60% EtOH extracts in mouse B16F10 melanoma cells 30
• 3.4.1. B16F10 cell viability with Parnassia palustris 60% EtOH extracts 30
• 3.4.2. Change of B16F10 cell morphology by Parnassia palustris 60% EtOH extracts treatment 32
• 3.4.3. Inhibition of melanin synthesis and tyrosinase activity by Parnassia palustris 60% EtOH extracts 34
• 3.4.4. Alterations in melanogenesis-associated protein levels, inducing MITF, TYR, TRP-1, and TRP-2 following treatment with Parnassia palustris 60% EtOH extracts 36
• 3.4.5. Transcriptional modulation of melanogenesis-related genes, inducing MITF, TRP-1, MC1R, Myo5a, and Rab27a by Parnassia palustris 60% EtOH extracts 39
• 4. Conclusion 43
• 5. References 45
• Abstract in Korean 52