慢性乙肝患者ERα基因SNPs和mRNA表达及与α干扰素疗效关联分析

Association of ERαgene single nucleotide polymorphisms and mRNA expression levels with therapeutic efficacy of interferon alfa in patients with chronic hepatitis B

目的在慢性乙型病毒性肝炎(chronic hepatitis b,CHB)的抗病毒治疗药物中,α干扰素因为具有很强的免疫调节作用,从而显著增强了该药的抗病毒疗效。但CHB患者的α干扰素抗病毒疗效个体差异大,已有报道雌激素受体α(estrogen receptor alpha,ERα)基因单核苷酸多态性(single nucleotide polymorphisms,SNPs)和mRNA表达水平与CHB易感性相关,本研究拟检测α干扰素治疗后CHB患者外周血单核细胞(peripheral blood mononuclear cells,PBMCs)ERα基因SNPs和mRNA表达水平,探讨ERα基因SNPs和mRNA水平与α干扰素疗效的关系。方法2014-01-03-2016-01-07,对76例接受α干扰素治疗的乙肝病毒e抗原(hepatitis b virus e antigen,HbeAg)阳性CHB患者进行随访,评估其抗病毒疗效。采用质谱分析法检测α干扰素治疗后第48周的患者及健康对照者PBMCs中的ERαPVUⅡ和XBaⅠ基因SNPs,采用荧光染料实时定量PCR法(SYBR Green RT-PCR)检测α干扰素治疗后第48周的患者及健康对照者PBMCs中ERαmRNA的表达水平。结果α干扰素治疗后第48周,ERαPVUⅡ基因的PP、Pp和pp基因型的病毒学应答率分别为52.4%、70.0%和42.9%,差异无统计学意义,χ2=3.762,P=0.152;HBeAg血清学转换率分别为61.5%、61.1%和57.1%,差异无统计学意义,χ2=0.083,P=0.959。ERαXBaⅠ基因的XX、Xx和xx基因型病毒学应答率分别为56.5%、57.9%和47.1%,差异无统计学意义,χ2=0.774,P=0.679;HBeAg血清学转换率分别为60.9%、60.0%和64.3%,差异无统计学意义,χ2=0.123,P=0.940。CHB患者PBMCs中的ERαmRNA表达水平为8.27±7.44,低于健康对照组的15.397±9.06,但差异无统计学意义,t=-1.177,P=0.233;在α-干扰素治疗后,病毒学应答组患者ERαmRNA水平较治疗前显著升高,t=-7.587,P<0.001,而无应答组患者的ERαmRNA表达水平较治疗前无显著变化,t=-2.296,P=0.051;HBeAg血清学转换组(t=-1.752,P=0.118)和无转换组(t=-1.089,P=0.308)ERαmR-NA水平在治疗后差异均无统计学意义。结论 ERαPVUⅡ和XBaⅠ基因SNPs与α-干扰素疗效无显著关联。通过检测PBMCs中ERαmRNA表达水平有无显著性提高,可以预测CHB患者的α-干扰素治疗效果。

OBJECTIVE To analyze the association of estrogen receptor alpha(ERα)gene single nucleotide polymorphisms(SNPs)and mRNA expression levels with thERαpeutic efficacy of interferon alfa(IFN-α)in patients with chronic hepatitis B(CHB).METHODS A total of 76 CHB patients with IFN-αtreatment were enrolled from January 3,2014 to January 7,2016.All patients were followed up and the efficacy of antiviral thERαpy was examined.SNPs of PVUⅡand XBaⅠloci in ERαgene were examined by the Sequence Mass ARRAY.ERαmRNA expression from periphERαl blood mononuclear cells(PBMCs)was measured by reverse transcription-polymERαse chain reaction(RT-PCR).RESULTS The 48-week virological response rates in patients with PP,Pp and pp genotype at PVUⅡlocus of ERαgene were 52.4%,70.0%and 42.9%,respectively.There were no statistically significant differences between them(χ^2=3.762,P=0.152).The 48-week HBeAg seroconversion rates in patients with PP,Pp,and pp genotype at PVUⅡlocus of ERαgene were 61.5%,61.1%,and 57.1%,respectively.There were no statistically significant differences between them(χ^2=0.083,P=0.959).The 48-week virological response rates in patients with XX,Xx,and xx genotype at XBaⅠlocus of ERαwere 56.5%,57.9%,and 47.1%,respectively.There were also no statistically significant differences between them(χ^2=0.774,P=0.679).Their HBeAg seroconversion rates were 60.9%,60.0%,and 64.3%,respectively.There were no statistically significant differences between them(χ^2=0.123,P=0.940).The ERαmRNA expression levels of PBMCs in patients with chronic hepatitis B were lower than that in healthy controls(8.27±7.44 vs 15.397±9.06,t=-1.177,P=0.233).The expression of ERαmRNA were significantly increased in patients with virological response after antiviral thERαpy(t=-7.587,P<0.001),while there was no significant change of expression levels of ERαmRNA in non-respondent patients before and after antiviral thERαpy(t=-2.296,P=0.051).No significant change of ERαmRNA level were found in patients with or without HBeAg serological conversion after treatment(t=-1.752,P=0.118 and t=-1.089,P=0.308,respectively).CONCLUSIONS SNPs of PVUⅡand XBaⅠloci in ERαgene are not significantly correlated with the thERαpeutic efficacy of IFN-α.The level of ERαmRNA from PBMCs can be an indicator for assessing the thERαpeutic efficacy of IFN-αin CHB patients.