Venner T J, Sauder D N, Feliciani C, Mckenzie R C
Division of Dermatology, University of Toronto, Sunnybrook Health Science Centre, Ontario, Canada.
Exp Dermatol. 1995 Jun;4(3):138-45. doi: 10.1111/j.1600-0625.1995.tb00237.x.
Ultraviolet radiation can induce the transcription and release of cytokines from keratinocytes (KC's). These cytokines have the potential to modulate local and systemic immunologic responses. In this paper we report that northern blotting showed that human KC and KC lines expressed a 1.2-1.4 kb transcript for the chemokine and melanoma growth-stimulatory protein, GRO-alpha and that ultraviolet B radiation (UVB) could upregulate the expression of GRO-alpha mRNA and protein in the KC line A431. The GRO-alpha gene response to UVB was maximal at 48h post-irradiation with 70 J/m2. Reverse transcription-polymerase chain reaction (RT-PCR) revealed a 4.5-fold increase in GRO-alpha mRNA over basal levels (p < 0.001). GRO-alpha protein was measured in the culture media by enzyme-linked immunosorbent assay (ELISA). Media from unirradiated cultures contained 1166 +/- 83 pg/ml GRO-alpha protein. After UVB, a time-dependent increase in GRO-alpha protein was seen in the culture media from 6-48h. At 48h post-irradiation the GRO-alpha protein content was 27583 +/- 678 pg/ml, or 23 times the basal level. This protein release could be inhibited by 70% when the cells were pre-incubated with 10 micrograms/ml interleukin-1 receptor antagonist (IL-1RA). We also show that another potent leukocyte chemoattractant, Interleukin-8 (IL-8), was induced in A431 cells by UVB. This induction of IL-8 mRNA began as early as 3h post-irradiation, when it reached twice basal levels (p < 0.05) and reached 4.5-fold basal levels at 48h post-irradiation (p < 0.005).(ABSTRACT TRUNCATED AT 250 WORDS)
紫外线辐射可诱导角质形成细胞(KC)转录并释放细胞因子。这些细胞因子有可能调节局部和全身免疫反应。在本文中,我们报告通过Northern印迹法显示,人KC和KC系表达趋化因子及黑色素瘤生长刺激蛋白GRO-α的1.2 - 1.4 kb转录本,且紫外线B辐射(UVB)可上调KC系A431中GRO-α mRNA和蛋白的表达。GRO-α基因对UVB的反应在照射后48小时、70 J/m2时达到最大。逆转录聚合酶链反应(RT-PCR)显示GRO-α mRNA比基础水平增加了4.5倍(p < 0.001)。通过酶联免疫吸附测定(ELISA)检测培养基中的GRO-α蛋白。未照射培养物的培养基中含有1166±83 pg/ml的GRO-α蛋白。UVB照射后,培养基中GRO-α蛋白在6 - 48小时呈时间依赖性增加。照射后48小时,GRO-α蛋白含量为27583±678 pg/ml,是基础水平的23倍。当细胞与10微克/毫升白细胞介素-1受体拮抗剂(IL-1RA)预孵育时,这种蛋白释放可被抑制70%。我们还表明,另一种强效白细胞趋化因子白细胞介素-8(IL-8)在A431细胞中也被UVB诱导。IL-8 mRNA的这种诱导最早在照射后3小时开始,此时达到基础水平的两倍(p < 0.05),并在照射后48小时达到基础水平的4.5倍(p < 0.005)。(摘要截短于250字)