免疫调节剂对人A375黑素瘤细胞黑素合成的影响

发布时间：2018-03-09 08:13

本文选题：人A375黑素瘤细胞　切入点：免疫调节剂　出处：《南华大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的：通过观察不同浓度的卡介菌多糖核酸（BCG）、胸腺肽对人A375黑素瘤细胞的生长、酪氨酸酶活性及黑素生成的影响，探讨免疫调节剂对治疗白癜风的可能作用机制。方法：人A375黑素瘤细胞用不同浓度的BCG、胸腺肽处理，用多巴氧化法测定酪氨酸酶活性的变化，通过MTT法观察它们对人A375黑色瘤细胞生长的影响，用NaOH裂解法测定黑素的含量。结果：1.两种药物对人A375黑素瘤细胞代谢的影响：与对照组相比：BCG的浓度为0.1ug/ml、0.5ug/ml、1ug/ml、2.5ug/ml、5ug/ml、10ug/ml时随着药物浓度的增加，细胞数目呈增加趋势（p＜0.05）,细胞生长旺盛。BCG的处理时间24h、48h及72h比较,48h处理时达到最佳促进细胞生长效果。而浓度为50ug/ml—300ug/ml胸腺肽处理人A375黑素瘤细胞24h时，，对细胞的代谢却没有影响（p＞0.05），当处理时间达到48h时，各个浓度处理组均表现出对人A375细胞的增殖促进作用，且该促进作用呈剂量依赖性。而当处理时间延长至72小时时，这种促进作用的效果有所降低。 2.两种药物对人A375黑素瘤细胞酪氨酸酶活性影响：两种药物均显示上调细胞的酪氨酸酶活性。其中BCG在48小时10ug/ml浓度时上调酪氨酸酶的活性最为明显（p＜0.05）,而胸腺肽在72小时300ug/ml浓度时显示处最佳上调酪氨酸酶活性效果(p＜0.05)，其中胸腺肽使酪氨酸酶的活性增加的效果优于BCG。 3.两种药物对人A375黑素瘤细胞黑素含量的影响：两种药物均显示上调细胞的黑素含量。其中BCG处理后黑素瘤细胞中黑素含量升高不明显(P0.05)。而胸腺肽促进细胞黑素含量的效果比较明显，且呈时间剂量依赖性关系，其中以300μg/ml处理72h作用最为明显黑素含量增加(P0.05)。结论：一定浓度的BCG和胸腺肽均可以促进人A375黑素细胞增殖，上调细胞酪氨酸酶活性，从而达到促进黑素合成的效果，且他们的促进效果都与处理的时间及剂量相关联。
[Abstract]:Aim: to investigate the effects of different concentrations of BCGN and thymosin on the growth, tyrosinase activity and melanin production of human A375 melanoma cells, and to explore the possible mechanism of immunomodulator in the treatment of vitiligo. Methods: human A375 melanoma cells were treated with different concentrations of BCG and thymosin. The activity of tyrosinase was determined by dopa oxidation method. The effects of tyrosinase activity on the growth of human A375 melanoma cells were observed by MTT method. The content of melanin was determined by NaOH lysis method. Results 1. The effect of two drugs on the metabolism of human A375 melanoma cells: compared with the control group, the concentration of 0. 1 ug- BCG was 0. 1 ugr / ml, 0. 5 ugr / ml, 2. 5 ugr / ml, and 5. 5 ugP. Ml + 10 g / ml, with the increase of drug concentration. The number of cells increased (p < 0.05). The treatment time of exuberant BCG was 24h / 48h and 72h respectively, and the best effect was achieved when treated with 50ugml-300ug-1 / ml thymosin for 24 h and 72 h, respectively, and at the concentration of 50ugml-300ug-1 / ml thymosin for 24 h, the cells were treated with A375 melanoma cells. However, there was no effect on cell metabolism. When the treatment time was 48 h, the proliferation of A375 cells was enhanced in a dose-dependent manner, and when the treatment time was up to 72 hours, the proliferation of A375 cells was increased in a dose-dependent manner, and when the treatment time was prolonged to 72 hours, the proliferation of A375 cells was increased in a dose-dependent manner. The effect of this promotion has been reduced. 2. Effects of two drugs on tyrosinase activity in human A375 melanoma cells: both drugs showed up-regulation of tyrosinase activity, in which BCG increased tyrosinase activity most significantly at the concentration of 10ugrml at 48 hours (p < 0.05), while thymosin increased the activity of tyrosinase in A375 melanoma cells (P < 0.05). The optimal up-regulation of tyrosinase activity was found at the concentration of 300ugr / ml for 72 hours (p < 0.05), and the effect of thymosin on tyrosinase activity was better than that of BCG. 3. Effects of two drugs on melanin content in human A375 melanoma cells: both drugs showed up-regulation of melanin content. The melanin content in melanoma cells treated with BCG was not significantly increased, while thymosin promoted melanin content in melanoma cells. The effect of content is obvious. In a dose-dependent manner, 300 渭 g / ml treatment for 72 h had the most obvious effect on the increase of melanin content (P 0.05). Conclusion: both BCG and thymosin can promote the proliferation of human A375 melanocytes and up-regulate the activity of tyrosinase.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R739.5

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