本文選題：人A375黑素瘤細(xì)胞　切入點(diǎn)：免疫調(diào)節(jié)劑　出處：《南華大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：通過(guò)觀察不同濃度的卡介菌多糖核酸（BCG）、胸腺肽對(duì)人A375黑素瘤細(xì)胞的生長(zhǎng)、酪氨酸酶活性及黑素生成的影響，探討免疫調(diào)節(jié)劑對(duì)治療白癜風(fēng)的可能作用機(jī)制。 方法：人A375黑素瘤細(xì)胞用不同濃度的BCG、胸腺肽處理，用多巴氧化法測(cè)定酪氨酸酶活性的變化，通過(guò)MTT法觀察它們對(duì)人A375黑色瘤細(xì)胞生長(zhǎng)的影響，用NaOH裂解法測(cè)定黑素的含量。 結(jié)果：1.兩種藥物對(duì)人A375黑素瘤細(xì)胞代謝的影響：與對(duì)照組相比：BCG的濃度為0.1ug/ml、0.5ug/ml、1ug/ml、2.5ug/ml、5ug/ml、10ug/ml時(shí)隨著藥物濃度的增加，細(xì)胞數(shù)目呈增加趨勢(shì)（p＜0.05）,細(xì)胞生長(zhǎng)旺盛。BCG的處理時(shí)間24h、48h及72h比較,48h處理時(shí)達(dá)到最佳促進(jìn)細(xì)胞生長(zhǎng)效果。而濃度為50ug/ml—300ug/ml胸腺肽處理人A375黑素瘤細(xì)胞24h時(shí)，，對(duì)細(xì)胞的代謝卻沒(méi)有影響（p＞0.05），當(dāng)處理時(shí)間達(dá)到48h時(shí)，各個(gè)濃度處理組均表現(xiàn)出對(duì)人A375細(xì)胞的增殖促進(jìn)作用，且該促進(jìn)作用呈劑量依賴性。而當(dāng)處理時(shí)間延長(zhǎng)至72小時(shí)時(shí)，這種促進(jìn)作用的效果有所降低。 2.兩種藥物對(duì)人A375黑素瘤細(xì)胞酪氨酸酶活性影響：兩種藥物均顯示上調(diào)細(xì)胞的酪氨酸酶活性。其中BCG在48小時(shí)10ug/ml濃度時(shí)上調(diào)酪氨酸酶的活性最為明顯（p＜0.05）,而胸腺肽在72小時(shí)300ug/ml濃度時(shí)顯示處最佳上調(diào)酪氨酸酶活性效果(p＜0.05)，其中胸腺肽使酪氨酸酶的活性增加的效果優(yōu)于BCG。 3.兩種藥物對(duì)人A375黑素瘤細(xì)胞黑素含量的影響：兩種藥物均顯示上調(diào)細(xì)胞的黑素含量。其中BCG處理后黑素瘤細(xì)胞中黑素含量升高不明顯(P0.05)。而胸腺肽促進(jìn)細(xì)胞黑素含量的效果比較明顯，且呈時(shí)間劑量依賴性關(guān)系，其中以300μg/ml處理72h作用最為明顯黑素含量增加(P0.05)。 結(jié)論：一定濃度的BCG和胸腺肽均可以促進(jìn)人A375黑素細(xì)胞增殖，上調(diào)細(xì)胞酪氨酸酶活性，從而達(dá)到促進(jìn)黑素合成的效果，且他們的促進(jìn)效果都與處理的時(shí)間及劑量相關(guān)聯(lián)。
[Abstract]:Aim: to investigate the effects of different concentrations of BCGN and thymosin on the growth, tyrosinase activity and melanin production of human A375 melanoma cells, and to explore the possible mechanism of immunomodulator in the treatment of vitiligo. Methods: human A375 melanoma cells were treated with different concentrations of BCG and thymosin. The activity of tyrosinase was determined by dopa oxidation method. The effects of tyrosinase activity on the growth of human A375 melanoma cells were observed by MTT method. The content of melanin was determined by NaOH lysis method. Results 1. The effect of two drugs on the metabolism of human A375 melanoma cells: compared with the control group, the concentration of 0. 1 ug- BCG was 0. 1 ugr / ml, 0. 5 ugr / ml, 2. 5 ugr / ml, and 5. 5 ugP. Ml + 10 g / ml, with the increase of drug concentration. The number of cells increased (p < 0.05). The treatment time of exuberant BCG was 24h / 48h and 72h respectively, and the best effect was achieved when treated with 50ugml-300ug-1 / ml thymosin for 24 h and 72 h, respectively, and at the concentration of 50ugml-300ug-1 / ml thymosin for 24 h, the cells were treated with A375 melanoma cells. However, there was no effect on cell metabolism. When the treatment time was 48 h, the proliferation of A375 cells was enhanced in a dose-dependent manner, and when the treatment time was up to 72 hours, the proliferation of A375 cells was increased in a dose-dependent manner, and when the treatment time was prolonged to 72 hours, the proliferation of A375 cells was increased in a dose-dependent manner. The effect of this promotion has been reduced. 2. Effects of two drugs on tyrosinase activity in human A375 melanoma cells: both drugs showed up-regulation of tyrosinase activity, in which BCG increased tyrosinase activity most significantly at the concentration of 10ugrml at 48 hours (p < 0.05), while thymosin increased the activity of tyrosinase in A375 melanoma cells (P < 0.05). The optimal up-regulation of tyrosinase activity was found at the concentration of 300ugr / ml for 72 hours (p < 0.05), and the effect of thymosin on tyrosinase activity was better than that of BCG. 3. Effects of two drugs on melanin content in human A375 melanoma cells: both drugs showed up-regulation of melanin content. The melanin content in melanoma cells treated with BCG was not significantly increased, while thymosin promoted melanin content in melanoma cells. The effect of content is obvious. In a dose-dependent manner, 300 渭 g / ml treatment for 72 h had the most obvious effect on the increase of melanin content (P 0.05). Conclusion: both BCG and thymosin can promote the proliferation of human A375 melanocytes and up-regulate the activity of tyrosinase.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R739.5

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