Baldea I, Mocan T, Cosgarea R
Department of Physiology, University of Medicine and Pharmacy Iuliu Hatieganu, Cluj-Napoca 400006, Romania.
Exp Oncol. 2009 Dec;31(4):200-8.
Melanocytes are producing melanin after UV irradiation as a defense mechanism. However, UV-induced damage is involved in melanoma initiation, depending on skin phototype. Melanocytes seem to be extremely susceptible to free radicals. Their main enzymatic antioxidants are superoxide dismutase and catalase.
To study how melanin synthesis modulates the activity of the oxidative stress defense enzymes and cell proliferation after UV induced cell damage.
Normal human melanocyte cultures from fair skin individuals were exposed to high levels of L-tyrosine and irradiated, with 20, 30, 40 mJ/cm2 UVA, and respective UVB. Proliferation was measured using a MTS assay; viability was assessed by trypan blue exclusion dye method. Spectrophotometrical methods were used to determine total melanin content, the enzymatic activity of tyrosinase, superoxide dismutase and catalase.
Tyrosine had a negative effect on proliferation, enhanced with time elapsed. Overall, UV irradiation decreased proliferation. UVA increased proliferation relative to UVB in the cultures exposed for a longer time to high (2 mM) tyrosine concentration. There were no proliferation differences between UVA and UVB irradiation in lower tyrosine concentration exposed melanocytes. Both, UV irradiation and tyrosine increased melanogenesis. Exposure of the melanocytes to increased levels of tyrosine in medium (0.5 mM and 1 mM) and UV irradiation enhanced the activity of superoxide dismutase and catalase. The enzymes showed a high activity rate in melanocytes while exposed for a short time to 2 mM tyrosine, but their activity was dramatically decreased with longer tyrosine exposure and UV irradiation.
Our data indicate that in low phototype melanocytes, melanogenesis, either following UV irradiation, or tyrosine exposure, especially in high concentrations, was detrimental for the cells by reducing the activity of catalase and superoxidedismutase, the natural antioxidants. UVA was more efficient in stimulating the activity of superoxide dismutase and catalase but also in depleting the reserves of the enzymatic defense against oxidative stress, especially catalase, than UVB. This physiologic response to UV light can be considered as an adjunctive risk factor for people with low phototype for developing a melanoma, when exposed to UV irradiation.
黑色素细胞在紫外线照射后会产生黑色素作为一种防御机制。然而,紫外线诱导的损伤与黑色素瘤的发生有关,这取决于皮肤光类型。黑色素细胞似乎对自由基极其敏感。它们主要的酶抗氧化剂是超氧化物歧化酶和过氧化氢酶。
研究紫外线诱导细胞损伤后黑色素合成如何调节氧化应激防御酶的活性和细胞增殖。
将来自皮肤白皙个体的正常人黑色素细胞培养物暴露于高浓度的L - 酪氨酸并进行照射,分别用20、30、40 mJ/cm²的UVA以及相应的UVB照射。使用MTS法测量增殖;通过台盼蓝排斥染色法评估活力。采用分光光度法测定总黑色素含量、酪氨酸酶、超氧化物歧化酶和过氧化氢酶的酶活性。
酪氨酸对增殖有负面影响,且随着时间推移而增强。总体而言,紫外线照射会降低增殖。在长时间暴露于高(2 mM)酪氨酸浓度的培养物中,相对于UVB,UVA增加了增殖。在低酪氨酸浓度暴露的黑色素细胞中UVA和UVB照射之间没有增殖差异。紫外线照射和酪氨酸都会增加黑色素生成。将黑色素细胞暴露于培养基中增加的酪氨酸水平(0.5 mM和1 mM)以及紫外线照射会增强超氧化物歧化酶和过氧化氢酶的活性。当短时间暴露于2 mM酪氨酸时,这些酶在黑色素细胞中显示出高活性率,但随着酪氨酸暴露时间延长和紫外线照射,它们的活性会显著降低。
我们的数据表明,在低光类型黑色素细胞中,紫外线照射或酪氨酸暴露后,尤其是高浓度时,黑色素生成通过降低过氧化氢酶和超氧化物歧化酶(天然抗氧化剂)的活性对细胞有害。与UVB相比,UVA在刺激超氧化物歧化酶和过氧化氢酶的活性方面更有效,但在消耗抗氧化应激的酶储备方面,尤其是过氧化氢酶,也更有效。当暴露于紫外线照射时,这种对紫外线的生理反应可被视为低光类型人群患黑色素瘤的一个辅助危险因素。