ISSN 1674-3865  CN 21-1569/R
主管：国家卫生健康委员会
主办：中国医师协会
　　　辽宁省基础医学研究所
　　　辽宁中医药大学附属医院
中国中西医结合儿科学

中国中西医结合儿科学 ›› 2023, Vol. 15 ›› Issue (5): 401-406.doi: 10.3969/j.issn.1674-3865.2023.05.008

• 实验论著 • 上一篇    下一篇

补气益精生血中药对人红系细胞组蛋白修饰酶的影响

陈珏璇, 徐方蔚, 卢焯明   

  1. 510120 广州，广州市妇女儿童医疗中心
  • 收稿日期:2023-08-09 出版日期:2023-10-25 上线日期:2023-10-25
  • 通讯作者: 卢焯明，E-mail:739835365@qq.com
  • 基金资助:
    广东省中医药局科研项目（20211301）；广州市妇女儿童医疗中心内部基金（NKE-PRE-2019-007）

The impact of Buqi Yijing Shengxue Chinese herbal medicines on histone-modifying enzymes in human erythroid cells

CHEN Juexuan, XU Fangwei, LU Zhuoming   

  1. Guangzhou Women and Children's Medical Center, Guangzhou 510120，China

  • Received:2023-08-09 Published:2023-10-25 Online:2023-10-25
  • Contact: LU Zhuoming，E-mail:739835365@qq.com

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摘要： 目的 观察补气益精生血中药对人红系K562细胞株组蛋白修饰酶的影响。方法 运用补气益精生血、补气生血、益精生血的中药分别以高、低浓度对K562细胞株进行诱导，并以丁酸钠作阳性对照。以实时荧光定量聚合酶链式反应(PCR)检测细胞γ珠蛋白基因基因mRNA水平，采用酶标仪以比色法检测细胞组蛋白脱乙酰基酶(HDAC1、HDAC2),组蛋白乙酰基转移酶(HAT)酶活性水平，PCR方法检测HDAC1、HDAC2、HAT基因mRNA水平，Western Blot方法检测HDAC1、HDAC2、HAT蛋白水平。结果 补气益精高剂量组、补气高剂量组、益精低剂量组中药及丁酸钠均显著提升了γ珠蛋白基因表达水平。补气益精高、低剂量组及益精高、低剂量组的细胞HAT酶活性水平均显著高于空白对照组，而丁酸钠组的HDAC1酶活性水平则显著低于空白对照组。对于HAT基因的mRNA水平，补气益精高、低剂量组，益精高剂量组显著高于空白对照组。Western Blot结果则只有补气益精高剂量组和益精高剂量组的HAT蛋白水平显著高于空白对照组。结论 补气益精生血中药可能通过上调HAT基因的表达进而促进红系细胞内γ珠蛋白基因启动子区组蛋白的乙酰化修饰，其诱导γ珠蛋白基因表达治疗儿童β地中海贫血的分子机制可能与调节组蛋白修饰酶有关。 

关键词: β地中海贫血, 中医药疗法, γ珠蛋白基因表达, 组蛋白修饰

Abstract: Objective This study aims to investigate the impact of Buqi Yijing Shengxue Chinese herbal medicines, which are known for their generating blood by tonifying Qi and benefiting essence properties, on histone-modifying enzymes in the human erythroid K562 cell line.Methods K562 cells were induced with high and low concentrations of several groups of Chinese herbal medicines, including Buqi Yijing Shengxue drugs, Buqi Shengxue drugs and Yijing Shengxue drugs for blood generating, and sodium butyrate was used as a positive control. The mRNA levels of the γ-globin gene were assessed using real-time fluorescence quantitative PCR. Additionally, cellular HDAC1, HDAC2, and HAT enzyme activity levels were analyzed using an enzyme marker by colorimetric detection methods. The mRNA levels of HDAC1, HDAC2, and HAT genes were determined using the PCR method, while protein levels of these genes were determined via Western Blot.Results Treatment with Chinese herbal medicine and sodium butyrate significantly increased the expression levels of the γ-globin gene in high-dose Qi-tonifying and essence-benefiting group, high-dose Qi-tonifying group, and low-dose essence-benefiting group. Cellular HAT enzyme activity levels were notably higher in high and low-dose Qi-tonifying and essence-benefiting groups, as well as in high and low-dose essence-benefiting groups than in blank control group,while HDAC1 enzyme activity levels in the sodium butyrate group were markedly lower than those recorded in the blank control group. HAT mRNA levels were notably higher in high and low-dose Qi-tonifying and essence-benefiting groups, as well as in high-dose essence-benefiting group than in blank control group. Western Blot outcomes demonstrated that only the HAT protein levels in the high-dose Qi-tonifying and essence-benefiting group and high-dose essence-benefiting group were considerably elevated compared to those in the blank control group.Conclusion The molecular mechanism of Buqi Yijing Shengxue Chinese herbal medicines in the induction of γ-globin in gene expression in the treatment of pediatric β-thalassemia may be related to the regulation of the histone-modifying enzyme, which may be mediated by up-regulating the expression of histone acetyltransferase HAT to enhance histone modification in erythroid cells.

Key words:

β-thalassemia, TCM therapy, γ-globin gene expression, Histone modification