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miRNA-548ah调控HDAC4影响HBV复制和表达及临床意义 被引量:5
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作者 王蔚 赵伟 +3 位作者 邢同京 肖丽 咸建春 尹有美 《中华医院感染学杂志》 CAS CSCD 北大核心 2018年第16期2401-2405,2410,共6页
目的探讨微小RNA-548ah(miRNA-548ah)调控组蛋白去乙酰化酶4(HDAC4)对乙型肝炎病毒(hepatitis B virus,HBV)复制和表达的影响及其在肝组织中的表达与相关性。方法选择2015年-2017年泰州市人民医院诊治的慢性HBV感染患者18例为研究对象,... 目的探讨微小RNA-548ah(miRNA-548ah)调控组蛋白去乙酰化酶4(HDAC4)对乙型肝炎病毒(hepatitis B virus,HBV)复制和表达的影响及其在肝组织中的表达与相关性。方法选择2015年-2017年泰州市人民医院诊治的慢性HBV感染患者18例为研究对象,其中慢性乙型肝炎(Chronic hepatitis B,CHB)患者9例,乙型肝炎肝硬化(LC)患者9例,荧光定量PCR检测三种不同肝癌细胞株及CHB患者肝组织中miRNA-548ah和HDAC4分子的表达,应用miRNA-548ah模拟剂和抑制剂转染Huh7细胞、HepG2细胞和HepG2,2,15细胞,检测细胞培养上清中HBsAg、HBeAg、HBV DNA的表达水平;Pearson相关分析CHB患者肝组织中miRNA548ah和HDAC4表达与临床指标的相关性。结果 miRNA-548ah和HDAC4在Huh7细胞、HepG2细胞和HepG2,2,15细胞的表达差异具有统计学意义(P<0.001);转染pHBV1.3质粒的HepG2细胞、Huh7细胞、HepG2,2,15细胞中,与对照组比较,miRNA-548ah模拟剂组HBsAg、HBeAg和HBV DNA的表达均升高,miRNA-548ah抑制剂组HBsAg、HBeAg和HBV DNA表达均降低(P<0.001);CHB患者肝组织中miRNA548ah的表达为(0.90±0.56),低于乙肝LC患者的表达(1.62±0.71)(P<0.05);CHB患者肝组织中miRNA548ah和HDAC4的表达呈负相关(r=-0.746,P=0.021),肝组织中HDAC4的表达水平与血清ALT的水平呈正相关(r=0.733,P=0.025)。结论 miRNA-548ah可通过靶向HDAC4调控乙型肝炎病毒的复制与表达,并且影响CHB的病情进展。 展开更多
关键词 miRNA-548ah HDAC4 乙型肝炎 慢性
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HepG2-hNTCP细胞模型的构建及在miRNA-548ah调控乙型肝炎病毒研究中的应用 被引量:1
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作者 邢同京 王学全 +2 位作者 田雄 李阿利 周妮 《中华医院感染学杂志》 CAS CSCD 北大核心 2021年第8期1154-1159,共6页
目的构建并鉴定表达人钠离子-牛磺胆酸共转运蛋白(hNTCP)的HepG2-hNTCP细胞模型,探讨微小RNA-548ah(miR-548ah)对乙型肝炎病毒复制和表达的影响。方法扩增重组慢病毒pWPI-hNTCP-Puro质粒,pWPI-hNTCP-Puro慢病毒感染HepG2细胞,聚合酶链反... 目的构建并鉴定表达人钠离子-牛磺胆酸共转运蛋白(hNTCP)的HepG2-hNTCP细胞模型,探讨微小RNA-548ah(miR-548ah)对乙型肝炎病毒复制和表达的影响。方法扩增重组慢病毒pWPI-hNTCP-Puro质粒,pWPI-hNTCP-Puro慢病毒感染HepG2细胞,聚合酶链反应(PCR)检测hNTCPmRNA的表达水平,共聚焦显微镜检测hNTCP蛋白定位。Lipofectamine转染miR-548ah激动剂、miR-548ah抑制剂及阴性对照至细胞系。酶联免疫吸附测定检测HBsAg和HBeAg的表达水平,荧光定量PCR检测HBVDNA的表达,应用SPSS 17.0对数据进行分析。结果hNTCP mRNA在HepG2hNTCP呈高水平表达。共聚焦荧光显微镜观察HepG2hNTCP中的hNTCP蛋白正常表达且主要定位在细胞膜上。与HepG2细胞比较,HBsAg和HBeAg在感染后HepG2-hNTCP细胞5天和7天后的表达水平存在差异,随培养天数呈上升趋势,具有统计学意义(P<0.01)。与转染阴性对照比较,HBsAg在转染miR-548ah激动剂组的表达水平升高,差异具有统计学意义(P<0.05);而HBeAg和HBVDNA的表达水平无明显变化,差异均无统计学意义。结论成功构建了表达hNTCP的HepG2-hNTCP细胞模型,对乙型肝炎病毒感染具有良好的易感性;miR-548ah可促进HBV感染HepG2-hNTCP细胞模型中HBsAg的表达。 展开更多
关键词 乙型肝炎病毒 钠离子-牛磺胆酸共转运蛋白 微小RNA-548ah HepG2-hNTCP细胞模型
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Association of T regulatory cells with natural course and response to treatment with interferon-α in patients with chronic hepatitis B infection 被引量:10
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作者 XU Hong-tao xing tong-jing +1 位作者 LI Hao YE Jun 《Chinese Medical Journal》 SCIE CAS CSCD 2012年第8期1465-1468,共4页
Background Regulatory T cell populations, particularly CD4+CD25+ T regulatory cells, have been implicated in the persistence of hepatitis B virus (HBV) infection. However, no clear relationship has been establishe... Background Regulatory T cell populations, particularly CD4+CD25+ T regulatory cells, have been implicated in the persistence of hepatitis B virus (HBV) infection. However, no clear relationship has been established between the frequency of CD4~CD25~ T regulatory cells in the peripheral blood and either the disease phases in the natural history of chronic HBV infection or in the response to interferon-a therapy. Methods In the present study, three different common markers of CD4+CD25+ T regulatory cells were used to determine the numbers of T regulatory cells in healthy controls and in patients with chronic HBV infection. Results No significant difference was found when samples were gated for CD25hi and CD25+FoxP3+ T cells. A significant correlation was found between the number of CD4+ Treg cells that gated with CD25+FoxP3+ and CD25+CD127low/- in healthy controls and in patients with chronic hepatitis B (CHB) (r=0.67, 0.59; P 〈0.01). The percentages of Treg cells were (8.56±2.01)% in asymptomatic carriers (Asc), (8.74±3.04)% in inactive HBsAg carriers, (10.7±2.93)% in CHB and (7.42±1.28)% in healthy controls (F=-11.1, P 〈0.001). The percentage of Treg cells in patients with CHB was higher than in asymptomatic HBV patients, inactive HBsAg carriers, or healthy controls (P 〈0.01). The proportion of CD4+CD25+CD127 low/T cells in patients who responded to interferon-(] was (11.9±3.3)%, (9.1±2.4)% and (9.0±2.9)% at baseline, week 12 and week 24 after treatment, respectively (Z=2.42, P〈0.05; Z=2.67, P〈0.01). Conclusions These results suggest that the proportion of the CD4+CD25+ regulatory T cells might be affected by the application of different markers in process to detect T regulatory cells. The frequency of Treg cells was increased in patients with CHB, which might be associated with the disease activity of these patients and contribute to prevention of extensive liver damage. A decline in Treg cells at week 12 of treatment might be associated with a better response to treatment with interferon-α. 展开更多
关键词 CD4+CD25+ regulatory T cells FOXP3 chronic hepatitis B INTERFERON-Α
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