他卡西醇对人表皮黑素细胞调控作用的研究

发布时间：2018-06-05 04:27

本文选题：他卡西醇 + 黑素细胞　；参考：《暨南大学》2012年硕士论文

【摘要】：目的探讨他卡西醇对体外培养的人表皮黑素细胞增殖、黏附、迁移的调控作用及其对c-kitmRNA相对表达量的影响。通过上述实验，为他卡西醇在白癜风复色治疗中的作用机制提供一定的理论及实验室依据。方法用不同浓度的他卡西醇干预体外培养的人表皮黑素细胞，采用XTT法分别检测培养24h、48h及72h后黑素细胞的增殖活性，用纤维连接蛋白包被细胞培养板检测培养72h后黑素细胞的黏附率，用Transwell微孔膜法检测培养24h后黑素细胞在纤维连接蛋白上的迁移情况，经RT-PCR法检测培养72h后黑素细胞c-kit mRNA相对表达量。结果 1.他卡西醇在浓度为10-10~10-6mol/l范围内对人表皮黑素细胞增殖无明显抑制作用。当浓度为10-8mol/l的他卡西醇与黑素细胞共同培养48h、72h，与对照组（不加药仅添加完全培养基）相比，能显著促进黑素细胞增殖，差异有统计学意义（P分别为0.029、0.044）。 2.他卡西醇浓度为10-8~10-7mol/l范围内与对照组（不加药仅添加完全培养基）相比，可显著促进人表皮黑素细胞在纤维连接蛋白（FN）上的黏附，差异有统计学意义（P分别为0.003、0.007）。 3.依据前面细胞增殖活性的结果，各浓度他卡西醇对人表皮黑素细胞无明显毒性反应，，选择共同培养24h的细胞进行后续的迁移实验。结果显示，浓度为10-9~10-8mol/l的他卡西醇与对照组（不加药仅添加完全培养基）相比，在培养24h时能显著促进黑素细胞迁移（P分别为0.003、0.000）。 4.经RT-PCR半定量分析结果表明，浓度为10-9~10-7mol/l的他卡西醇与对照组（不加药仅添加完全培养基）相比，在培养72h均能显著增加黑素细胞c-Kit mRNA的相对表达量，差异有统计学意义（P均为0.000）。结论 1.他卡西醇能够促进体外培养的人表皮黑素细胞的增殖，增强黑素细胞在FN上的黏附能力及诱导黑素细胞在FN上的迁移作用； 2.他卡西醇可部分上调人表皮黑素细胞c-kit mRNA的相对表达量，且c-kit mRNA表达的变化与黑素细胞黏附及迁移作用有一定相关性，他卡西醇可能通过上调c-kit表达从而促进人表皮黑素细胞的黏附、迁移。
[Abstract]:Purpose To investigate the effects of tacrazol on the proliferation, adhesion and migration of cultured human epidermal melanocytes and its effect on the relative expression of c-kitmRNA. These experiments provide a theoretical and laboratory basis for the mechanism of tacacicol in the treatment of vitiligo. Method Human epidermal melanocytes were treated with different concentrations of tacoxiol in vitro. The proliferation activity of melanocytes was detected by XTT assay after cultured for 24 h or 72 h, respectively. The adhesion rate of melanocytes after 72 h culture was detected with fibronectin coated cell culture plate, and the migration of melanocytes on fibronectin after 24 h culture was detected by Transwell microporous membrane method. The relative expression of c-kit mRNA in melanocytes was detected by RT-PCR assay after 72 h culture. Result 1. Tacarbazol had no inhibitory effect on the proliferation of human epidermal melanocytes in the range of 10-10~10-6mol/l. When the concentration of 10-8mol/l was cocultured with melanocytes for 48 h or 72 h, the proliferation of melanocytes was significantly increased compared with the control group (without adding only complete medium), with a significant difference (P = 0.029, 0.044, respectively). 2. Compared with the control group (the control group), the concentration of tacoxiol in the range of 10-8~10-7mol/l could significantly promote the adhesion of human epidermal melanocytes to fibronectin (FN). The difference was statistically significant (P = 0.003, P = 0.007, respectively). 3. According to the results of proliferative activity of the former cells, there was no obvious toxic effect on human epidermal melanocytes in each concentration of tacoxiol, and the cells co-cultured for 24 hours were selected for subsequent migration test. The results showed that tacoxiol at the concentration of 10-9~10-8mol/l could significantly promote the migration of melanocytes in 24 hours compared with the control group (without adding only complete medium) (P = 0.003 ~ 0.000). 4. The results of RT-PCR semi-quantitative analysis showed that the relative expression of c-Kit mRNA in melanocytes was significantly increased at 72 h after cultured with 10-9~10-7mol/l, compared with that of the control group (without adding only complete medium), and the difference was statistically significant (P = 0.000). Conclusion 1. Tacrazol could promote the proliferation of human epidermal melanocytes in vitro, enhance the adhesion of melanocytes on FN and induce the migration of melanocytes on FN. 2. The relative expression of c-kit mRNA in human epidermal melanocytes was partly upregulated by tacrazol, and the change of c-kit mRNA expression was related to the adhesion and migration of melanocytes. Tacassicol may promote the adhesion and migration of human epidermal melanocytes by upregulating the expression of c-kit.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R758.41

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