發(fā)布時(shí)間：2018-10-14 08:24

【摘要】：目的:通過觀察不同濃度的氫化可的松、醋酸潑尼松龍、地塞米松磷酸鈉對人A375黑素瘤細(xì)胞的生長、酪氨酸酶活性以及黑素生成的影響,探討皮質(zhì)類固醇治療白癜風(fēng)的可能作用機(jī)制。 方法:用不同濃度的氫化可的松、醋酸潑尼松龍、地塞米松磷酸鈉處理人A375黑素瘤細(xì)胞,通過MTT比色法觀察它們對人A375黑素瘤細(xì)胞生長的影響,用多巴氧化法測定酪氨酸酶活性的變化,用NaOH裂解法測定黑素的含量。 結(jié)果:1.三種藥物對人A375黑素瘤細(xì)胞代謝的影響:與對照組相比,氫化可的松的濃度為10~(-6)～10~(-5) M時(shí)隨著藥物濃度的增加,細(xì)胞數(shù)目呈增加趨勢(p 0.05),細(xì)胞生長旺盛,而濃度為10~(-9)～10~(-5) M的醋酸潑尼松龍、地塞米松磷酸鈉對人A375黑素瘤細(xì)胞的代謝卻沒有影響(p0.05)。 2.三種藥物對人A375黑素瘤細(xì)胞酪氨酸酶活性的影響:3種藥物均顯示上調(diào)細(xì)胞的酪氨酸酶活性。其中地塞米松磷酸鈉在10~(-5) M～10~(-8) M濃度時(shí)顯示均可上調(diào)酪氨酸酶的活性(p 0.05),而氫化可的松和醋酸潑尼松龍僅在10~(-5) M和10~(-6)M濃度時(shí)才顯示上調(diào)酪氨酸酶活性(p 0.05),其中10~(-5) M地塞米松磷酸鈉作用最明顯,使酪氨酸酶的活性增加最多。 3.三種藥物對人A375黑素瘤細(xì)胞黑素含量的影響:3種藥物均顯示上調(diào)細(xì)胞的黑素含量。其中地塞米松磷酸鈉在10~(-5) M～10~(-8) M濃度時(shí)均可上調(diào)黑素含量(p 0.05),而氫化可的松和醋酸潑尼松龍僅在10~(-5) M和10~(-6) M濃度時(shí)才顯示上調(diào)黑素含量(p 0.05),其中10~(-5) M地塞米松磷酸鈉作用最明顯,使黑素含量增加最多。 結(jié)論:一定濃度的氫化可的松可以促進(jìn)人A375黑素瘤細(xì)胞增殖;一定濃度的氫化可的松、醋酸潑尼松龍、地塞米松磷酸鈉皆可以上調(diào)酪氨酸的活性,促進(jìn)黑素的合成。
[Abstract]:Aim: to observe the effects of hydrocortisone, prednisolone acetate and dexamethasone phosphate on the growth, tyrosinase activity and melanogenesis of human A375 melanoma cells. To explore the possible mechanism of corticosteroid in the treatment of vitiligo. Methods: human A375 melanoma cells were treated with different concentrations of hydrocortisone, prednisolone acetate and dexamethasone phosphate. The effects of hydrocortisone, prednisolone acetate and dexamethasone sodium phosphate on the growth of human A375 melanoma cells were observed by MTT colorimetry. The activity of tyrosinase was determined by dopa oxidation method and the content of melanin was determined by NaOH pyrolysis method. The result is 1: 1. Effects of three drugs on Metabolism of Human A375 melanoma cells: compared with the control group, when the concentration of hydrocortisone was 10 ~ (-6) ~ 10 ~ (-5) M, the number of cells increased with the increase of drug concentration (p 0.05). However, 10 ~ (-9) ~ 10 ~ (-5) M prednisolone acetate and dexamethasone sodium phosphate had no effect on the metabolism of human A375 melanoma cells (p0.05). Effects of three drugs on tyrosinase activity in human A375 melanoma cells: all three drugs showed up-regulated tyrosinase activity. Dexamethasone sodium phosphate could upregulate tyrosinase activity at 10 ~ (-5) M ~ (-8) M concentration (p 0.05), while hydrocortisone and prednisolone acetate showed up-regulated tyrosinase activity only at 10 ~ (-5) M and 10 ~ (-6) M concentrations (p 0.05). The effect of 10 ~ (-5) M dexamethasone sodium phosphate was the most obvious. Tyrosinase activity increased the most. 3. Effects of three drugs on melanin content in human A375 melanoma cells: all three drugs showed up-regulation of melanin content. Dexamethasone sodium phosphate upregulated melanin content (p0.05) at 10 ~ (-5) M ~ (-8) M concentration, while hydrocortisone and prednisolone acetate only increased melanin content (p 0.05) at the concentrations of 10 ~ (-5) M and 10 ~ (-6) M, of which 10 ~ (-5) M dexamethasone increased melanin content (p0.05). Sodium phosphate has the most obvious effect. The content of melanin increased the most. Conclusion: hydrocortisone at a certain concentration can promote the proliferation of human A375 melanoma cells, and hydrocortisone, prednisolone acetate and dexamethasone phosphate can up-regulate tyrosine activity and promote melanin synthesis.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R739.5

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