中药抗白灵霜对小鼠B16黑素瘤细胞代谢影响的实验研究

发布时间：2018-02-28 14:38

本文关键词： 中药抗白灵霜白癜风 B16小鼠黑素瘤细胞酪氨酸酶黑素合成　出处：《南京中医药大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的：研究中药抗白灵霜(Kang Bai Ling,简称KBLSS)对体外培养的小鼠B16黑素瘤细胞增殖、酪氨酸酶活性以及黑素含量的影响。探讨药物与其影响1316黑素瘤细胞代谢的相关因素,阐明其治疗白癜风的作用机制和原理。方法：体外培养小鼠B16黑素瘤细胞,使5个不同剂量组的药物与细胞共同孵育,以不加细胞的DMEM培养基作为空白组,以单纯细胞作为对照组,用细胞学以及酶学等手段观察药物对B16小鼠黑素瘤细胞增殖、酪氨酸酶活性及黑素合成的影响。采用四甲基偶氮唑蓝比色法(MTT法)测定小鼠黑素瘤细胞的增殖,采用左旋多巴氧化速率法测定对酪氨酸酶活性的影响,采用NaOH裂解法检测黑素含量。结果： 1.抗白灵霜水提浸膏能促进B16黑素瘤细胞增殖。将结果与对照组相比,5个剂量组均有显著性差异(P0.05)。其中3个剂量组(15－25mg／ml)有明显增殖,5组随药物浓度的升高,呈剂量依赖性。在浓度较低时(5－lOmg／ml)表现为轻微的抑制作用。 2.抗白灵霜水提浸膏对酪氨酸酶有明显的激活作用。据结果所示,在药物与细胞共同培养24h后,与48h,72h相比激活作用最为明显(P0.05),且尤其表现在低浓度组上(5－15mg／ml).与对照组相比,5个剂量组均有显著上调酪氨酸酶活性的作用(P0.05),5个剂量组(5－25mg／ml)均随药物浓度的升高,呈剂量依赖性 3.抗白灵霜水提浸膏能促进B16黑素瘤细胞的黑素生成,提高黑素含量。与对照组相比,5个剂量组均有明显促黑素生成的作用(P0.05),5个剂量组随药物浓度的升高,略有降低,考虑在较低浓度(5mg／ml)时更有利于黑素的合成。 4.抗白灵霜醇提浸膏能促进B16黑素瘤细胞增殖。将结果与对照纠相比,5个剂量组均有显著性差异(P0.05)。5个剂量组均有促进细胞增殖的作用(P0.05),5个剂量组(5－25mg／ml)均随药物浓度的升高,呈剂量依赖性。 5.抗白灵霜醇提浸膏对酪氨酸酶有明显的激活作用。据结果所示,在药物与细胞共同培养48h后,与24h,72h相比激活作用最为明显(P0.05)。与对照组相比,5个剂量组均有显著上调酪氨酸酶活性的作用(P0.05),5个剂量组(5－25mg／ml)均随药物浓度的升高,呈剂量依赖性 6.抗白灵霜醇提浸膏能促进B16田黑素瘤细胞的黑素生成,提高黑素含量。与对照组相比,5个剂量组均有明显促黑素生成的作用(P0.05),5个剂量组均有促进细胞增殖的作用(P0.05),5个剂量组(5-25mg/ml)均随药物浓度的升高,呈剂量依赖性结论：中药抗白灵霜的水提浸膏与醇提浸膏均能促进B16黑素瘤细胞增殖,提高酪氨酸酶活性,进而促进黑素合成。总体上随着药物浓度的增加,以上作用均有增强的趋势。
[Abstract]:Objective: to study the effects of anti-bailing cream Kang Bai Ling( KBLSSfor short) on the proliferation, tyrosinase activity and melanin content of B16 melanoma cells cultured in vitro, and to explore the related factors affecting the metabolism of 1316 melanoma cells. To elucidate the mechanism and principle of the treatment of vitiligo. Methods: murine B16 melanoma cells were cultured in vitro. Five different dosages of drugs were incubated with the cells. The DMEM medium without cells was used as the blank group and the simple cells as the control group. The effects of drugs on the proliferation, tyrosinase activity and melanin synthesis of B16 mouse melanoma cells were observed by cytological and enzymatic methods. The proliferation of mouse melanoma cells was determined by MTT assay with tetramethylazolyl blue colorimetry. The activity of tyrosinase was determined by levodopa oxidation rate method and melanin content was detected by NaOH pyrolysis method. Results:. 1. The water extract extract of Kangbailing cream could promote the proliferation of B16 melanoma cells. Compared with the control group, there were significant differences among the five dose groups (P 0.05). In a dose-dependent manner, 5-lOmg / ml showed a slight inhibitory effect at lower concentrations. 2. The water extract extract of Kangbailing cream can activate tyrosinase obviously. Compared with the control group, the activation effect of P0.05 was the most obvious, especially in the low concentration group. Compared with the control group, the activity of tyrosinase was upregulated significantly in all the five dose groups (P0.05N). The concentration of 5-25mg / ml in the five dose groups increased with the increase of the drug concentration in a dose-dependent manner. 3. The water extract extract of Kangbailing cream could promote melanin production and increase the content of melanin in B16 melanoma cells. Compared with the control group, all the five dose groups had the effect of melanin production (P 0.05), and the five dose groups decreased slightly with the increase of drug concentration. At a lower concentration of 5 mg / ml, it is more favorable for the synthesis of melanin. 4. The alcohol extract extract of Kangbailing cream could promote the proliferation of B16 melanoma cells. Compared with the control group, there were significant differences in the results among the five dose groups, the five dose groups all had the effect of promoting cell proliferation (P0.05%), and the 5 dose groups (5-25 mg / ml) increased the concentration of B16 melanoma cells. In a dose-dependent manner. 5. The alcohol extract of Kangbailing cream can activate tyrosinase obviously. According to the results, after 48 hours of co-culture of the drug and the cells, Compared with the control group, the activity of tyrosinase was upregulated significantly in all the five dose groups, and 5-25 mg / ml in a dose-dependent manner with the increase of the concentration of the drug. 6. The alcohol extract extract of Kangbailing cream can promote melanin production in B16 melanoma cells. Compared with the control group, all the five dose groups had the effect of melanin production, the five dose groups all had the effect of promoting cell proliferation, and the five dose groups had the effect of promoting cell proliferation in a dose-dependent manner with the increase of drug concentration (5-25 mg / ml). Conclusion: both aqueous extract and alcohol extract of anti-Bailing cream can promote the proliferation of B16 melanoma cells, increase tyrosinase activity, and then promote melanin synthesis.
【学位授予单位】：南京中医药大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R285.5;R739.5

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