人参皂甙Rb1对人表皮黑素细胞黑素生成的影响

发布时间：2018-02-04 17:39

本文关键词： 人参皂甙Rb1 黑素细胞黑素生成　出处：《大连医科大学》2011年硕士论文　论文类型：学位论文

【摘要】：目的:皮肤中黑素含量是决定皮肤颜色的主要因素之一。黑素细胞产生黑素,并将其输送到周围的角质形成细胞,从而影响皮肤的颜色。黑素细胞生成黑素的不足可以导致皮肤色素减退或脱失,促进黑素细胞的黑素生成是治疗色素脱失性疾病的策略之一。酪氨酸酶是黑素细胞产生黑素的关键酶。近年研究发现,酪氨酸酶主要受小眼畸形相关转录因子(MITF)的调控,而MITF的表达又受到上游蛋白如cAMP应答元件结合蛋白(CREB)的影响。中药人参曾被用于白癜风等脱色素性疾病的治疗,而Rb1是中药人参的主要有效成分之一。近年来有报道人参皂甙Rb1具有抗健忘、抗衰老、抗氧化、抗凋亡,促进神经纤维形成并保持其功能、等多重作用。黑素细胞和神经细胞在胚胎发生中同源,但人参皂甙Rb1对黑素细胞黑素生成的作用尚未见报道。本文研究人参皂甙Rb1对人表皮黑素细胞黑素生成的影响及其作用机制,以探讨其用于治疗色素脱失性皮肤病的前景。方法:将黑素细胞进行原代培养、传代,选取第2-5代黑素细胞进行实验,采用MTT法测定细胞增殖情况,氧化多巴法测定酪氨酸酶活性,氢氧化钠裂解法测定黑素生成,免疫蛋白印记法测定酪氨酸酶和小眼畸形相关转录因子(MITF)的表达及cAMP应答元件结合蛋白(CREB)的激活。结果:加入25-100μM人参皂甙Rb1作用72h后,黑素细胞活力无明显影响,细胞酪氨酸酶活性和黑素含量呈浓度依赖性增强;100μM人参皂甙Rb1作用72h后,酪氨酸酶、MITF蛋白表达明显增加,CREB明显激活。10μM的蛋白激酶A(PKA)抑制剂H-89预处理细胞,可明显抑制人参皂甙Rb1引起的CREB激活、酪氨酸酶及MITF蛋白表达的增加,进而抑制人参皂甙Rb1引起的酪氨酸酶活性作用增强及黑素合成增加。结论:人参皂甙Rb1可促进正常人黑素细胞的黑素合成和酪氨酸酶活性。PKA/CREB/MITF/酪氨酸酶信号通路参与了人参皂甙Rb1的促黑素生成机制。
[Abstract]:Objective: the content of melanin in skin is one of the main factors that determine the skin color. Melanocytes produce melanin and transport it to the surrounding keratinocytes. Thus affecting the color of the skin. The lack of melanocyte production melanin can lead to hypopigmentation or loss of skin. Promoting melanogenesis in melanocytes is one of the strategies for the treatment of dystrocytic diseases. Tyrosinase is the key enzyme for melanocytes to produce melanin. Tyrosinase is mainly regulated by MITF, a transcription factor associated with microocular malformation. The expression of MITF is affected by upstream proteins such as cAMP response element binding protein (cAMP). Ginseng has been used in the treatment of depigmented diseases such as vitiligo. In recent years, ginsenoside Rb1 has been reported to have anti-amnesia, anti-aging, anti-oxidation, anti-apoptosis, promote the formation of nerve fibers and maintain its function. Melanocytes and nerve cells are homologous in embryogenesis. However, the effect of ginsenoside Rb1 on melanogenesis of melanocytes has not been reported. The effect of ginsenoside Rb1 on melanogenesis of human epidermal melanocytes and its mechanism have been studied in this paper. To explore its application in the treatment of pigmented dermatosis. Methods: melanocytes were cultured and subcultured. The proliferation of melanocytes was measured by MTT assay and tyrosinase activity was determined by oxidative dopa method. The formation of melanin was determined by sodium hydroxide pyrolysis method. The expression of tyrosinase and MITF and activation of cAMP response element binding protein (CREBB) were detected by immuno protein imprinting. Results: after treated with 25-100 渭 M ginsenoside Rb1 for 72 h, the activity of melanocytes was not affected, and the activity of tyrosinase and the content of melanin were increased in a dose-dependent manner. After treated with ginsenoside Rb1 for 72 hours, the protein expression of tyrosinase was significantly increased. The activation of CREB induced by ginsenoside Rb1 was significantly inhibited by the pretreatment of H89, a protein kinase (PKA) inhibitor of .10 渭 M, which was activated by CREB. The expression of tyrosinase and MITF protein increased, which inhibited the increase of tyrosinase activity induced by ginsenoside Rb1 and the increase of melanin synthesis. Conclusion:. Ginsenoside Rb1 can promote melanin synthesis and tyrosinase activity in normal human melanocytes. PKA / CREB- MITF / tyrosinase signaling pathway is involved in the melanogenesis mechanism of ginsenoside Rb1. .
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R285.5;R758.5

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