组蛋白乙酰化抑制哮喘易感基因ADRB2的表达

Histone Acetylation Inhibits the Expression of Asthma Susceptibility Gene ADRB2.

目的研究组蛋白去乙酰化酶抑制剂(histone deacetylase inhibitors,HDACi)及组蛋白乙酰化转移酶(histone acetyltransferase,HAT)调控的组蛋白乙酰化修饰对β2肾上腺素受体(β2-drenergic receptor,ADRB2)基因的影响。方法将ADRB2启动子质粒(pADRB2-1918)转染到人正常支气管上皮细胞(BEAS-2B)细胞后,分别加入丁酸钠(sodium butyrate,SoB)和曲古抑菌素A(trichostatin A,TSA),将pADRB2-1918质粒和腺病毒E1A相关的300kDa蛋白(p300)质粒共转染到BEAS-2B细胞,使BEAS-2B细胞中组蛋白乙酰化,利用双荧光素酶报告基因检测经SoB、TSA及p300处理后BEAS-2B细胞中ADRB2启动子活性。向BEAS-2B细胞中分别加入SoB和TSA,或向BEAS-2B细胞中转染p300质粒,使BEAS-2B细胞中组蛋白乙酰化,利用实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)检测BEAS-2B细胞中ADRB2mRNA的相对表达,利用Western blot法检测BEAS-2B细胞中ADRB2蛋白的表达。结果在BEAS-2B细胞中分别加入SoB、TSA和转染p300质粒,参与BEAS-2B细胞中组蛋白乙酰化,发现BEAS-2B细胞中ADRB2启动子活性、mRNA和蛋白的表达均较对照组下降(P<0.05),差异均有统计学意义。结论组蛋白乙酰化可下调哮喘易感基因ADRB2的表达。

Objective To study the effect of acetylation modification regulated by histone deacetylase inhibitors(HDACi)and histone acetyltransferase(HAT)onβ2-adrenergic receptor(ADRB2)gene.Methods ADRB2 promoter plasmid(pADRB2-1918)was transfected into BEAS-2B cells,and sodium butyrate(SoB)and trichostatin A(TSA)were added respectively.Adenovirus E1A-related kDa protein(p300)plasmid and PADRB2-1918 plasmid were co-transfected into BEAS-2B cells to acetylate histone in BEAS-2B cells.The activity of ADRB2 promoter in BEAS-2B cells treated with SoB,TSA,and p300 was detected by dual-luciferase gene reporter assay.SoB and TSA were added to BEAS-2B cells,or p300 plasmid was transfected into BEAS-2B cells to acetylate histone in BEAS-2B cells.The relative expression of ADRB2mRNA in BEAS-2B cells was detected by quantitative real-time PCR(qRT-PCR),and the expression of ADRB2 protein in BEAS-2B cells was detected by Western blot.Results SoB,TSA,and p300 plasmid were added into BEAS-2B cells,respectively,which participated in histone acetylation in BEAS-2B cells.We found that the activities of ADRB2 promoter,mRNA,and protein expressions in BEAS-2B cells were lower than those in the control group(P<0.05),and the differences were statistically significant.Conclusion Histone acetylation can inhibit the expression of ADRB2 in asthma susceptible genes.