欧前胡素、异欧前胡素，芍药苷对人表皮黑色细胞生物活性及迁移的影响

发布时间：2018-05-09 20:23

本文选题：欧前胡素 + 异欧前胡素　；参考：《泸州医学院》2013年硕士论文

【摘要】：目的：通过体外建立模拟正常生理状态的人表皮黑素细胞培养体系，观察白芷的两种主要单体欧前胡素、异欧前胡素和白芍的主要单体芍药苷对黑素细胞生物活性及迁移的影响，从而探讨这三种中药单体对黑素细胞的作用机制，为色素性疾病的治疗提供新的思路。方法：1、体外培养模拟皮肤正常生理状态的人表皮黑素细胞；2、检测不同浓度的三种中药单体（欧前胡素、异欧前胡素、芍药苷）对黑素细胞增殖活性的影响（MTT法）；3、检测不同浓度的三种中药单体（欧前胡素、异欧前胡素、芍药苷）对黑素细胞酪氨酸酶活性的影响（L-DOPA法）；4、检测不同浓度的三种中药单体（欧前胡素、异欧前胡素、芍药苷）对黑素细胞黑素合成的影响（NaOH法）；5、采用Transwell小室检测三种中药单体（欧前胡素、异欧前胡素、芍药苷）对黑素细胞迁移的影响。以上全部数据采用SPSS统计软件17.0版进行分析。结果：1、欧前胡素组在浓度为20μg/mL和50μg/mL时；异欧前胡素组在浓度为5μg/mL、10μg/mL、20μg/mL时；芍药苷组在浓度为5μg/mL、10μg/mL、20μg/mL、50μg/mL时均显示出显著的促进黑素细胞增殖的作用。各组中药单体在100μg/mL时并未对黑素细胞增殖产生明显的抑制作用，细胞仍显示较好的生长。各组中药单体对黑素细胞增殖的促进作用均是低浓度时呈浓度依赖性增强，而较高浓度时反而作用力度下降，由实验结果可见，10μg/mL为芍药苷和补骨脂素的最佳作用浓度；20μg/mL为欧前胡素和异欧前胡素的最佳作用浓度。其中5μg/mL、10μg/mL、20μg/mL浓度时的异欧前胡素组促细胞增殖能力强于同浓度的补骨脂素组。2、欧前胡素组在浓度为20μg/mL、50μg/mL时；异欧前胡素组在浓度为10μg/mL、20μg/mL时；芍药苷组在10μg/mL、20μg/mL、50μg/mL浓度时；均显示出上调黑素细胞酪氨酸酶活性的作用，并且其作用强度与同浓度的补骨脂素组相似。各组中药单体对调节黑素细胞酪氨酸酶活性的作用也呈现出低浓度时剂量依赖性上调，而高浓度时上调能力逐渐下降。10μg/mL为异欧前胡素和芍药苷的最佳作用浓度；20μg/mL为欧前胡素和补骨脂素的最佳作用浓度。3、欧前胡素组在浓度为2.5μg/mL、5μg/mL、10μg/mL、20μg/mL、50μg/mL、100μg/mL时均未显示出对黑素合成的促进作用；异欧前胡素组仅在20μg/mL时显著促进黑素合成；芍药苷组在浓度为20μg/mL、50μg/mL时显著的促进黑素合成。异欧前胡素组、芍药苷组、补骨脂素组促进黑素合成的作用均是低浓度时剂量依赖性上调，高浓度时下降。20μg/mL为异欧前胡素和补骨脂素的最佳作用浓度；50μg/mL为芍药苷的最佳作用浓度。4、10μg/mL、20μg/mL、50μg/mL的欧前胡素组均无明显促进黑素细胞迁移的作用；异欧前胡素组在这三个浓度时促黑素细胞迁移的作用呈现浓度依赖性加强，并于20μg/mL和50μg/mL时显示出极显著的促进黑素细胞迁移的作用，与空白对照组比较在统计学上具有显著性差异（P0.01）；芍药苷组在20μg/mL时极显著的促进黑素细胞迁移，与空白对照组比较在统计学上具有显著性差异（P0.01）；补骨脂素组（阳性对照组）在10μg/mL、20μg/mL、50μg/mL三个浓度均显著的促进黑素细胞迁移，其中20μg/mL时其作用强度最高；50μg/mL浓度组的异欧前胡素和补骨脂素均极显著的促进黑素细胞迁移，与空白对照组比较在统计学上具有显著性差异（P0.01），并且此时两者作用强度相当。结论：1、欧前胡素、异欧前胡素、芍药苷均能促进黑素细胞的增殖，且在较高浓度（100μg/mL）时对细胞无明显毒性作用，证实了这三种中药单体的安全性。并且异欧前胡素促黑素细胞增殖活性能力强于补骨脂素。2、欧前胡素、异欧前胡素、芍药苷均能上调黑素细胞酪氨酸酶的活性，，且作用强度与补骨脂素相似。3、异欧前胡素和芍药苷均能促进黑素细胞的黑素合成，而欧前胡素未检测出促进黑素合成的作用。4、20μg/mL、50μg/mL组的异欧前胡素和20μg/mL组的芍药苷均能显著地促进黑素细胞迁移，而10μg/mL、20μg/mL、50μg/mL浓度组的欧前胡素均无明显促进黑素细胞迁移的作用，由此推测异欧前胡素单体是白芷中促进黑素细胞迁移的主要活性成分，芍药苷单体是白芍中促进黑素细胞迁移的主要活性成分。50μg/mL组的异欧前胡素和补骨脂素促黑素细胞迁移的能力相当。
[Abstract]:Objective: To observe the effects of two main monomeric monosonoids, isomer and Paeoniae paeoniflorin on the bioactivity and migration of melanocytes by establishing a human epidermal melanocyte culture system in vitro to simulate normal physiological state, and to explore the mechanism of action of these three kinds of monomers on melanocytes. The treatment of vegetative diseases provides new ideas. Methods: 1, human epidermal melanocytes are cultured in vitro to simulate the normal physiological state of the skin. 2, the effects of three kinds of traditional Chinese medicine monomers on the proliferation of melanocytes (MTT method); 3, three different concentrations of traditional Chinese medicine (erhu) The effect of peptide, ISO, paeoniflorin on the activity of tyrosinase in melanocytes (L-DOPA); 4, the effects of three kinds of Chinese herbs on melanin synthesis of melanocytes (NaOH method) were detected by different concentrations of Chinese herbs (eomerhu, ISO, and paeoniflorin); 5, using the Transwell chamber to detect three kinds of Chinese medicine monomers (eomerhu, isooin, peony, Paeonia lactiflora The effect of glucoside on melanocyte migration. All the above data were analyzed by the 17 edition of SPSS software. Results: 1, when the concentration was 20 mu g/mL and 50 mu g/mL, the concentration of ISO euclixin group was 5 u g/mL, 10 mu g/mL, 20 mu g/mL, and the concentration of paeoniflorin in the concentration was 5 mu g /mL, 10 u g/mL, 20 u g/mL, 50 g/mL. The proliferation of melanocytes was promoted. The cell proliferation of melanocytes was not inhibited significantly at 100 g/mL. The cells still showed good growth. The experimental results showed that 10 mu g/mL was the best concentration of paeoniflorin and psoralen; 20 g/mL was the best concentration of eomorsin and isooctin. The proliferation ability of ISO euclixin group at 5 mu g/mL, 10 mu g/mL and 20 mu g/mL was stronger than that of the same concentration of.2 in the same concentration of bonalin group, and the concentration of oealin group was 20 mu g/mL, 50 mu. At the time of g/mL, when the concentration was 10 mu g/mL and 20 u g/mL, the group of paeoniflorin showed the role of up regulation of tyrosinase activity in melanocytes at 10 u g/mL, 20 mu g/mL and 50 micron g/mL, and its action intensity was similar to that of the same concentration of psoralen group. At low concentration, the dose dependence was also up, while the up-regulation ability of the high concentration decreased gradually.10 mu g/mL as the best concentration of isoeuin and paeoniflorin; 20 micron g/mL was the best concentration of.3, and the oealin group was 2.5 u g/mL, 5 u g/mL, 10 mu g/mL, 20 mu g/mL, 50 u g/mL, 100 micron g/mL. The promoting effect of melanin synthesis was not shown; the group of isooctin was significantly promoted by melanin synthesis at 20 g/mL; paeoniflorin group promoted melanin synthesis at a concentration of 20 mu and 50 mu g/mL. The effect of the group of isoeuin, paeoniflorin group, and psoralen group on melanin synthesis was up to a dose dependent up and high concentration in low concentration. The decrease of.20 mu g/mL was the best concentration of isoeuin and psoralen, the best concentration of 50 mu g/mL was.4,10 mu g/mL, 20 g/mL, and 50 uh g/mL had no significant effect on the migration of melanocytes; the effect of the migration of melanocytes at these three concentrations showed a concentration dependent manner. The effect on melanocyte migration was significantly increased at 20 g/mL and 50 u g/mL, which was significantly different from that in the blank control group (P0.01); the group of paeoniflorin significantly promoted the migration of melanocytes at 20 u g/mL, and had significant differences compared with the blank control group (P0.01). Psoralen group (positive control group) significantly promoted melanocyte migration at 10 g/mL, 20 g/mL and 50 micron g/mL, of which 20 mu g/mL had the highest strength, and the 50 mu g/mL concentration group of isooctin and psoralen all significantly promoted melanocyte migration, compared with the blank control group, the difference was statistically significant difference. The difference (P0.01), and at the same time, has the same action intensity. Conclusion: 1, eulein, isoeulin and paeoniflorin all can promote the proliferation of melanocytes and have no obvious toxic effects on the cells at a high concentration of 100 mu, which confirms the safety of the three monomers of the Chinese medicine. Psoralen.2, ocuarinin, isooctin and paeoniflorin can all up regulate the activity of melanocyte tyrosinase, and the action intensity is similar to.3, isooctin and paeoniflorin all can promote melanin synthesis in melanocytes, but the effect of eucartin on melanin synthesis is not detected in.4,20 mu g/mL, and the 50 mu g/mL group of isooeus Paeoniflorin in group 20 and 20 mu could significantly promote melanin cell migration, while 10 mu g/mL, 20 mu g/mL, and 50 mu g/mL concentration group did not significantly promote melanocyte migration. Thus, it is suggested that ISO oculin monomer is the main active component in the promotion of melanocyte migration in Angelica dahurica, and paeoniflorin monomer is the promoting melanin in Paeonia lactiflora The main active components of cell migration were isoisoaltein and Psoralen in.50 g/mL group.

【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R758.4;R392.12

【参考文献】