细菌基序对黑素细胞及前列腺素E_2的影响

The influence of bacteria motif on melanocytes and in prostaglandim E_2

目的:研究细菌基序(CpG-ODN)对黑素细胞形态、黑素生成及前列腺素E2(PGE2)的影响。方法:用CpG-ODN处理体外培养的B16黑素瘤细胞和melan-α黑素细胞,分别用MTT法、多巴氧化法、NaOH裂解法和ELISA法检测B16细胞活力、酪氨酸酶活性、黑素含量以及上清中PGE2的水平,另外,观察CpG-ODN干预后melan-α细胞形态的变化。结果:干预24h后,5μg/mlCpG-ODN促进B16细胞增殖,10μg/ml对细胞增殖的影响与空白对照组无差异,而15μg/mlCpG-ODN则抑制细胞增殖。48h后,不同浓度CpG-ODN组的细胞增殖率趋于稳定,与正常对照组间差异无统计学意义(P>0.05)。10μg/mlCpG-ODN提高酪氨酸酶活性和黑素生成,作用近似于α-MSH(P>0.05)。另外,此浓度CpG-ODN作用48h后的细胞上清液中PGE2含量也有所升高,但与对照组差异无统计学意义(P>0.05),推测为检测时间滞后所致。10μg/mlCpG-ODN作用于melan-α细胞24h后,细胞树突数目增加,并有二级以致三级树突出现。结论:CpG-ODN作用于鼠黑素细胞的适宜浓度为10μg/ml,此浓度对黑素细胞的作用有时间依赖性。该实验为CpG-ODN作用于正常人黑素细胞的研究奠定了基础。

Objective：To investigate the effects of bacteria motif,namely CpG oligodeoxynucleotide（CpG-ODN）on morphology and melanogenesis production of and prostaglandin E2（PGE2）in melanocytes.Methods：The cultured B16 melanoma cells and melan-α melanocytes in vitro were treated with the CpG-ODN.The B16 cell viability,tyrosinase activity,melanin content and the level of PGE2 in the culture supernatants was measured by MTT assay,oxidation of L-dopa,NaOH assay and ELISA assay respectively.In addition,the morphological changes of melan-α cells were observed after the CpG-ODN was added.Results：The CpG-ODN at the concentration of 5 μg/ml promoted the proliferation of B16 cells,10 μg/ml with little effects,and 15 μg/ml with depressant effects on the cell proliferation after drugs intervention for 24 h.After 48 h treating,the cell proliferation rates in various groups became stable and were similar to each other,without significant difference between groups at different concentrations of CpG-ODN（P〉0.05）.The tyrosinase activity and melanin synthesis were promoted by 10 μg/ml CpG-ODN,whose effects resembled that of 25 nmol/ml α-MSH（P〉0.05）.In addition,10 μg/ml CpG-ODN increased the level of PGE2 in the culture supernatants,but no statistical significance compared with the control group（P〉0.05）.It supposed that due to lagging of detection time.The cytodendrites of melan-α cells treated with 10 μg/ml CpG-ODN for 24 h were elongated and enriched,we could see the second and even tertiary dendrites appearing.Conclusions：The optimal concentration of CpG-ODN for stimulating mouse melanocytes is 10 μg/ml,and its effects are in a time-dependent manner.This study is a primary research about CpG-ODN stimulating normal human melanocytes.