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1. 辽宁中医药大学1. 经济管理学院公共管理教研室
2. 附属第三医院
3. 基础医学院中医基础理论教研室
4. 学生工作处
5. 教学实验中心生物技术实验室,沈阳,110847
纸质出版日期:2014,
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马月丹, 张 燚, 王继慧, 等. 绞股蓝总皂苷对紫外线照射的人皮肤细胞原癌基因表达的影响[J]. 重庆医科大学学报, 2014,38(3):360-363.
Ma Yuedan, Zhang Yi, Wang Jihui, et al. Effect of gypenosides on expression of proto-oncogene in skin cells following ultraviolet irradiation[J]. Journal of Chongqing Medical University, 2014,38(3):360-363.
目的:探讨绞股蓝总皂苷对紫外线照射的人皮肤相关原癌基因c-Fos和c-Jun表达的影响。方法:制备空白大鼠血清和绞股蓝含药血清。将人永生化角质形成细胞(human immortal skin keratinocytes,HaCaT)40瓶随机分为 4 组(A~D组),每组 10 瓶,将人皮肤成纤维细胞(human skin fibroblast,HSF)60 瓶随机分为 6 组(Ⅰ~Ⅵ组),每组10瓶。分别用中波紫外线(ultra-violet B,UVB)对 HaCaT 细胞 B~D 组、用长波紫外线(ultraviolet A,UVA)对 HSF 细胞Ⅱ~Ⅵ 组进行照射。将 HaCaT 4组细胞培养上清液分别加入HSF细胞的 Ⅲ~Ⅵ组。最终制成用于检测的6组细胞,分别为空白对照组(Ⅰ组)、UVA模型组(Ⅱ组)、Ha-CaT 培养上清液A组(Ⅲ组)、HaCaT 培养上清液B组(Ⅳ组)、 HaCaT 培养上清液C组(Ⅴ组)、HaCaT 培养上清液D组(Ⅵ组)。采用反转录聚合酶链反应(reverse transcription-PRC,RT-PCR)和蛋白免疫印迹(Western blot)方法检测Ⅰ~Ⅵ 组细胞c-Fos和c-Jun基因和蛋白的表达水平。结果:与Ⅰ组比较,Ⅱ组细胞c-Fos和c-Jun基因和蛋白的表达水平明显上调(各组P值均为0.000);与Ⅱ组比较,Ⅳ组c-Fos和c-Jun 基因和蛋白表达均上调(Pc-Fos基因=0.016、Pc-Fos蛋白=0.001、Pc-Jun基因=0.005、Pc-Jun蛋白=0.000),而 Ⅲ 组变化不明显;与Ⅳ组比较,Ⅵ组c-Fos和c-Jun基因和蛋白表达均下调(各组P值均为0.000),而Ⅴ组变化不明显(Pc-Fos基因=0.081、Pc-Fos蛋白=0.088、Pc-Jun基因=0.109、Pc-Jun蛋白=0.026)。结论:经UVB照射的HaCaT细胞分泌细胞因子促进经UVA照射的HSF细胞原癌基因c-Fos和c-Jun 基因和蛋白的表达。绞股蓝总皂苷对UVB照射HaCaT细胞培养上清液作用的UVA照射HSF细胞中原癌基因c-Fos和c-Jun的表达具有抑制作用。
Objective:To research the effect of gypenosides-containing serum on the expression of proto-oncogene c-Fos and c-Jun in ultraviolet irradiated cells. Methods:Blank serum and gypenosides-containing serum were made. Human immortal skin keratinocytes (HaCaT) were divided into 4 groups(A-D),10 bottles in each group. Human skin fibroblast(HSF) were divided into 6 groups(Ⅰ-Ⅵ),10 bottles in each group. B,C,D groups of irradiated HaCaT and Ⅱ,Ⅲ,Ⅳ,Ⅴ,Ⅵ groups of HSF underwent UVB and UVA radiation. Supernatant of 4 groups HaCaT was put into HSF Ⅲ-Ⅵ groups in turn. Six groups for measurement were made:blank control group(Ⅰgroup),UVA model group(Ⅱgroup),HaCaT supernatant A group(Ⅲ group),HaCaT supernatant B group(Ⅳ group),HaCaT supernatant C group(Ⅴgroup),HaCaT supernatant D group(Ⅵ group). Expression levels of c-Fos and c-Jun mRNA and protein in each group were measured by RT-PCR and Western blot. Results:c-Fos and c-Jun mRNA and protein expression was significantly higher in groupⅡ than in groupⅠ(all P =0.000). c-Fos and c-Jun mRNA and protein expression was significantly higher in group Ⅳ than in group Ⅱ(Pc-Fos mRNA=0.016,Pc-Fos protein=0.001,Pc-Jun mRNA=0.005,Pc-Jun protein=0.000). Changes in group Ⅲ were not significant. c-Fos and c-Jun mRNA and protein expression was significantly lower in group Ⅵ than in group Ⅳ(all P =0.000). Changes in group Ⅴ were not significant(Pc-Fos mRNA=0.081,Pc-Fos protein=0.088,Pc-Jun mRNA=0.109,Pc-Jun protein=0.026). Conclusions:The UVB irradiated HaCaT cell secrete cytokine to increase c-Fos and c-Jun mRNA and protein expression in UVA irradiated cell HSF. Gypenosides can inhibit c-Fos and c-Jun mRNA and protein expression of UVA irradiated HSF cells which are affected by UVB irradiated HaCaT supernatant.
绞股蓝总皂苷紫外线照射皮肤细胞原癌基因c-Fosc-Jun
gypenosidesultraviolet irradiationskin cellsproto-oncogenec-Fosc-Jun
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