Shi Mei-Jie, Xiao Huan-Ming, Xie Yu-Bao, Jiang Jun-Min, Zhao Peng-Tao, Cai Gao-Shu, Li Ying-Xian, Li Sheng, Zhang Chao-Zhen, Cao Min-Ling, Chen Qu-Bo, Tan Zhi-Jian, Gao Heng-Jun, Chi Xiao-Ling
The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou 510120, China.
Guangzhou University of Chinese Medicine, Guangzhou 510006, China.
Evid Based Complement Alternat Med. 2020 Oct 31;2020:5956940. doi: 10.1155/2020/5956940. eCollection 2020.
The aim of this study was to determine if microRNA (miRNA) expression is different among chronic hepatitis B (CHB) patients with early liver fibrosis classified according to traditional Chinese medicine (TCM) syndromes. Eighteen CHB-fibrosis patients and 12 CHB patients without fibrosis were enrolled. The CHB-fibrosis group included 9 patients with the TCM syndrome of Ganyu Pixu Xueyu (GYPXXY), characterized by liver stagnation, spleen deficiency, and blood stasis, and 9 patients with the TCM syndrome of Qixu Xueyu (QXXY), characterized by deficiency of qi, blood, and blood stasis. Agilent miRNA microarray was performed first in liver specimens to determine whether miRNA expression is different in patients with these two TCM syndromes of CHB-fibrosis. Gene Ontology (GO) analysis and KEGG analysis were applied to determine the roles of the differentially expressed miRNAs. QRT-PCR was performed to validate the Agilent miRNA microarray results. Compared with GYPXXY patients, 6 differentially expressed miRNAs were upregulated (miR-144-5p, miR-18a-5p, miR-148b-3p, miR-654-3p, miR-139-3p, and miR-24-1-5p) and 1 was downregulated (miR-6834-3p) in QXXY patients. According to qRT-PCR data, miR-144-5p and miR-654-3p were confirmed as upregulated in CHB-liver fibrosis patients compared to CHB patients without fibrosis, whereas the other 4 miRNAs were not significantly different. More importantly, miR-654-3p was confirmed to be significantly upregulated in QXXY patients compared with values in GYPXXY patients, whereas no significant difference was found in miR-144-5p. Moreover, the pathways of central carbon metabolism in cancer and cell cycle related to miR-654-3p and the target genes of PTEN and ATM were found to be different between QXXY patients and GYPXXY patients. These results indicate that there are different miRNAs, pathways, and target genes between QXXY patients and GYPXXY patients. However, due to the limited sample, whether miR-654-3p and the target genes PTEN and ATM could be molecular markers to differentiate TCM syndromes could not be established.
本研究旨在确定根据中医证候分类的慢性乙型肝炎（CHB）早期肝纤维化患者中，微小RNA（miRNA）表达是否存在差异。纳入18例CHB纤维化患者和12例无纤维化的CHB患者。CHB纤维化组包括9例肝瘀脾虚血瘀证（GYPXXY）的患者，其特征为肝郁、脾虚、血瘀，以及9例气血瘀证（QXXY）的患者，其特征为气虚、血虚、血瘀。首先在肝脏标本中进行安捷伦miRNA微阵列检测，以确定这两种CHB纤维化中医证候患者的miRNA表达是否存在差异。应用基因本体（GO）分析和京都基因与基因组百科全书（KEGG）分析来确定差异表达miRNA的作用。进行定量逆转录聚合酶链反应（QRT-PCR）以验证安捷伦miRNA微阵列的结果。与GYPXXY患者相比，QXXY患者中有6种差异表达的miRNA上调（miR-144-5p、miR-18a-5p、miR-148b-3p、miR-654-3p、miR-139-3p和miR-24-1-5p），1种下调（miR-6834-3p）。根据qRT-PCR数据，与无纤维化的CHB患者相比，CHB肝纤维化患者中miR-144-5p和miR-654-3p被证实上调，而其他4种miRNA无显著差异。更重要的是，与GYPXXY患者相比，QXXY患者中miR-654-3p被证实显著上调，而miR-144-5p未发现显著差异。此外，发现QXXY患者和GYPXXY患者中与miR-654-3p相关的癌症中心碳代谢途径和细胞周期以及PTEN和ATM的靶基因存在差异。这些结果表明，QXXY患者和GYPXXY患者之间存在不同的miRNA、途径和靶基因。然而，由于样本量有限，无法确定miR-654-3p以及靶基因PTEN和ATM是否可作为区分中医证候的分子标志物。
