本文關(guān)鍵詞： 中藥抗白靈霜 白癜風(fēng) B16小鼠黑素瘤細(xì)胞 酪氨酸酶 黑素合成　出處：《南京中醫(yī)藥大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：研究中藥抗白靈霜(Kang Bai Ling,簡(jiǎn)稱KBLSS)對(duì)體外培養(yǎng)的小鼠B16黑素瘤細(xì)胞增殖、酪氨酸酶活性以及黑素含量的影響。探討藥物與其影響1316黑素瘤細(xì)胞代謝的相關(guān)因素,闡明其治療白癜風(fēng)的作用機(jī)制和原理。 方法：體外培養(yǎng)小鼠B16黑素瘤細(xì)胞,使5個(gè)不同劑量組的藥物與細(xì)胞共同孵育,以不加細(xì)胞的DMEM培養(yǎng)基作為空白組,以單純細(xì)胞作為對(duì)照組,用細(xì)胞學(xué)以及酶學(xué)等手段觀察藥物對(duì)B16小鼠黑素瘤細(xì)胞增殖、酪氨酸酶活性及黑素合成的影響。采用四甲基偶氮唑藍(lán)比色法(MTT法)測(cè)定小鼠黑素瘤細(xì)胞的增殖,采用左旋多巴氧化速率法測(cè)定對(duì)酪氨酸酶活性的影響,采用NaOH裂解法檢測(cè)黑素含量。 結(jié)果： 1.抗白靈霜水提浸膏能促進(jìn)B16黑素瘤細(xì)胞增殖。將結(jié)果與對(duì)照組相比,5個(gè)劑量組均有顯著性差異(P0.05)。其中3個(gè)劑量組(15－25mg／ml)有明顯增殖,5組隨藥物濃度的升高,呈劑量依賴性。在濃度較低時(shí)(5－lOmg／ml)表現(xiàn)為輕微的抑制作用。 2.抗白靈霜水提浸膏對(duì)酪氨酸酶有明顯的激活作用。據(jù)結(jié)果所示,在藥物與細(xì)胞共同培養(yǎng)24h后,與48h,72h相比激活作用最為明顯(P0.05),且尤其表現(xiàn)在低濃度組上(5－15mg／ml).與對(duì)照組相比,5個(gè)劑量組均有顯著上調(diào)酪氨酸酶活性的作用(P0.05),5個(gè)劑量組(5－25mg／ml)均隨藥物濃度的升高,呈劑量依賴性 3.抗白靈霜水提浸膏能促進(jìn)B16黑素瘤細(xì)胞的黑素生成,提高黑素含量。與對(duì)照組相比,5個(gè)劑量組均有明顯促黑素生成的作用(P0.05),5個(gè)劑量組隨藥物濃度的升高,略有降低,考慮在較低濃度(5mg／ml)時(shí)更有利于黑素的合成。 4.抗白靈霜醇提浸膏能促進(jìn)B16黑素瘤細(xì)胞增殖。將結(jié)果與對(duì)照糾相比,5個(gè)劑量組均有顯著性差異(P0.05)。5個(gè)劑量組均有促進(jìn)細(xì)胞增殖的作用(P0.05),5個(gè)劑量組(5－25mg／ml)均隨藥物濃度的升高,呈劑量依賴性。 5.抗白靈霜醇提浸膏對(duì)酪氨酸酶有明顯的激活作用。據(jù)結(jié)果所示,在藥物與細(xì)胞共同培養(yǎng)48h后,與24h,72h相比激活作用最為明顯(P0.05)。與對(duì)照組相比,5個(gè)劑量組均有顯著上調(diào)酪氨酸酶活性的作用(P0.05),5個(gè)劑量組(5－25mg／ml)均隨藥物濃度的升高,呈劑量依賴性 6.抗白靈霜醇提浸膏能促進(jìn)B16田黑素瘤細(xì)胞的黑素生成,提高黑素含量。與對(duì)照組相比,5個(gè)劑量組均有明顯促黑素生成的作用(P0.05),5個(gè)劑量組均有促進(jìn)細(xì)胞增殖的作用(P0.05),5個(gè)劑量組(5-25mg/ml)均隨藥物濃度的升高,呈劑量依賴性 結(jié)論：中藥抗白靈霜的水提浸膏與醇提浸膏均能促進(jìn)B16黑素瘤細(xì)胞增殖,提高酪氨酸酶活性,進(jìn)而促進(jìn)黑素合成。總體上隨著藥物濃度的增加,以上作用均有增強(qiáng)的趨勢(shì)。
[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.
【學(xué)位授予單位】：南京中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R285.5;R739.5

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