丹参酮ⅡA对中波紫外线照射HaCaT细胞中GADD45α和激活蛋白-1及信号通路分子磷酸化水平的影响  

Effect of tanshinone ⅡA on the expressions of GADD45α,activator protein-1 and signaling pathway phosphorylation level in HaCaT cells induced by ultraviolet B irradiation

作　　者：韩昭[1] 高艳玲 李志锋[1] 燕霞[1] 李显平[1] 刘保国[1] 李小静[1] 刘志军[1] HAN Zhao;GAO Yan-ling;LI Zhi-feng;YAN Xia;LI Xian-ping;LIU Bao-guo;LI Xiao-jing;LIU Zhi-jun(Department of Dermatology, Affiliated Hospital of Hebei University of Engineering, Handan 056002, Chin)

机构地区：[1]河北工程大学附属医院皮肤科,河北邯郸056000 [2]邯郸市中心医院急诊科,河北邯郸056000

出　　处：《中华实用诊断与治疗杂志》2018年第4期323-326,共4页Journal of Chinese Practical Diagnosis and Therapy

基　　金：河北省科技计划项目(15277721D)

摘　　要：目的探讨丹参酮ⅡA对中波紫外线照射的HaCaT细胞中生长阻滞与DNA损伤基因45α(growth arrest and DNA damage-induced 45α,GADD45α)、激活蛋白-1(activator protein-1,AP-1)表达和信号通路分子磷酸化水平的影响。方法 HaCaT细胞采用中波紫外线照射后分为对照组和1mg/L丹参酮组、2mg/L丹参酮组、4mg/L丹参酮组,对照组加入等体积培养液,1mg/L丹参酮组、2mg/L丹参酮组、4mg/L丹参酮组分别加入1、2、4mg/L丹参酮处理48h。采用实时荧光定量PCR法检测4组细胞GADD45α、AP-1 mRNA表达情况,采用Western blot法检测4组细胞GADD45α、AP-1、细胞外调节蛋白激酶(extracellular regulated protein kinases,ERK)、c-Jun氨基末端激酶(Jun N-terminal kinase,JNK)、丝裂原活化蛋白激酶p38抗体(p38mitogen-activated protein kinase,p38MAPK)、蛋白激酶B(protein kinase B,Akt)蛋白表达情况。结果处理48h后,1mg/L丹参酮组、2mg/L丹参酮组和4mg/L丹参酮组AP-1mRNA(0.828±0.026、0.733±0.035、0.600±0.042)和GADD45αmRNA(0.791±0.041、0.708±0.039、0.569±0.006)表达均低于对照组(1.001±0.052、1.002±0.087)(P<0.05),4mg/L丹参酮组低于1mg/L丹参酮组和2mg/L丹参酮组,2mg/L丹参酮组低于1mg/L丹参酮组(P<0.05);1mg/L丹参酮组、2mg/L丹参酮组和4mg/L丹参酮组AP-1蛋白(0.889±0.149、0.599±0.044、0.435±0.078)、GADD45α蛋白(0.634±0.052、0.431±0.072、0.264±0.062)、ERK蛋白(0.457±0.068、0.380±0.041、0.306±0.025)、JNK蛋白(0.506±0.042、0.387±0.058、0.367±0.040)、p38 MAPK蛋白(0.697±0.083、0.493±0.047、0.371±0.012)和Akt蛋白(0.935±0.158、0.730±0.100、0.487±0.012)表达均低于对照组(1.019±0.158、0.706±0.039、0.662±0.063、0.589±0.051、0.704±0.052、1.086±0.100),4mg/L丹参酮组低于1 mg/L丹参酮组和2 mg/L丹参酮组,2 mg/L丹参酮组低于1 mg/L丹参酮组(P<0.05)。结论丹参酮ⅡA可抑制中波紫外线照射的HaCaT细胞表达GADD45α、AP-1和信号通路分子磷酸化水平。Objective To investigate the effect of tanshinone lI A on the expressions of growth arrest and DNA damageinduced 45α （GADD45α）, activator protein-1 （AP-1） and the related signal pathway phosphorylation level in HaCaT cells induced by ultraviolet B （UVB） irradiation. Methods HaCaT cells were divided into control group and 1, 2 and 4 mg/L tanshinone groups after UVB irradiation. Control group was added the equivalent volume of culture medium, and 1, 2 and 4 mg/L tanshinone groups were added 1, 2 and 4 mg/L tanshinone for 48 h. The expressions of GADD45α and AP-1 mRNA were detected by real-time PCR, and the expressions of GADD45α, AP-1, extracellular regulated protein kinases （ERK）, Jun N-terminal kinase （JNK）, p38 mitogen-activeted protein kinase （p38 MAPK） and protein kinase （Akt） were detected by Western blot technique. Results After 48 h, the mRNA levels of AP-1 （0. 828±0. 026, 0. 733±0. 035, 0. 600±0. 042） and GADD45α （0. 791±0. 041, 0. 708±0. 039, 0. 569±0. 006） in 1, 2 and 4 mg/L tanshinone groups were significantly lower than those in control group （1. 001±0. 052, 1. 002±0. 087） （P〈0.05）, in 4 mg/L tanshinone group than those in 1 mg/L tanshinone group and 2 mg/L tanshinone group, and in 2 mg/L tanshinone group than those in 1 mg/L tanshinonegroup （P〈0.05）. The protein levels of AP-1 （0.889±0.149, 0.599±0.044, 0.435±0.078）, GADD45α （0.634±0.052, 0.431±0. 072, 0. 264±0. 062）, ERK （0. 457±0. 068, 0. 380±0. 041, 0. 306±0. 025）, JNK （0. 506±0. 042, 0. 387±0. 058, 0. 367±0. 040）, p38 MAPK （0. 697±0. 083, 0. 493±0. 047, 0. 371±0. 012） and Akt （0. 935±0. 158, 0. 730±0. 100, 0. 487±0. 012） in 1, 2 and 4 mg/L tanshinone groups were significantly lower than those in control group （1. 019±0. 158, 0. 706±0. 039, 0. 662±0. 063, 0. 589±0. 051, 0. 704±0. 052, 1. 086±0. 100）, in 4 mg/L tanshinone group than those in 1 mg/L tanshinone group and 2 mg/L tanshinone group, and in 2 mg/L tanshinone group than thos

关 键 词：丹参酮ⅡA 中波紫外线 HACAT细胞 生长阻滞与DNA损伤基因45α 激活蛋白-1 

分 类 号：R285[医药卫生—中药学]