慢性乙型肝炎患者血清外泌体mir-122、mir-146a表达与HBV DNA载量的相关性

摘要/Abstract

摘要： 目的研究慢性乙型肝炎(CHB)患者血清外泌体microRNA-122(mir-122)、microRNA-146a(mir-146a)表达与HBV DNA的相关性。方法收集2018年5月至2019年12月南方医科大学深圳医院收治的CHB患者285例,依据荧光定量PCR(RT-qPCR)检测血清外泌体HBV DNA,分为低载量组(HBV DNA<1×10⁵拷贝/mL)、中载量组(1×10⁵拷贝/mL≤HBV DNA≤1×10⁷拷贝/mL)和高载量组(HBV DNA>1×10⁷拷贝/mL),每组95例。另取同期门诊正常健康体检者血清标本95份为对照组。采用RT-qPCR检测血清外泌体mir-122、mir-146a表达水平,采用化学发光法检测血清肝纤维化指标水平,采用Pearson相关性分析血清外泌体mir-122、mir-146a表达水平与HBV DNA载量及肝纤维化指标的相关性。结果与对照组比较,低、中、高载量组血清外泌体HBV DNA 载量及mir-122血清肝纤维化指标透明质酸酶(HA)、Ⅲ型前胶原(PC Ⅲ)、Ⅳ型胶原(C Ⅳ)、层粘连蛋白(LN)水平均依次增加,mir-146a水平依次降低(P<0.05),血清外泌体mir-122、mir-146a水平呈负相关(r=-0.765,P<0.05),mir-122与HBV DNA 载量、HA、PC Ⅲ、C Ⅳ、LN水平均呈正相关(r= 0.773、0.764、0.699、0.665、0.697,P<0.05),mir-146a与HBV DNA 载量、HA、PC Ⅲ、C Ⅳ、LN水平均呈负相关(r=-0.840、-0.779、-0.731、-0.691、-0.738,P<0.05)。结论 CHB患者血清外泌体mir-122上调、mir-146a水平下调,且与HBV DNA 载量密切相关,可能拮抗影响HBV DNA 复制及肝纤维化程度,与CHB发生发展有关。

关键词: 慢性乙型肝炎, microRNA-122, microRNA-146a, 乙型肝炎病毒, 外泌体

Abstract: Objective To study on the association of microRNA-122 (mir-122) and microRNA-146a (mir-146a) levels in serum exosomes of chronic hepatitis B (CHB) patients with hepatitis B viral DNA (HBV DNA) load. Methods 285 CHB patients were divided into low viral load group (HBV DNA < 1 ×10⁵ copy/ml), medium viral load group (1 ×10⁵ copy/ml ≤ HBV DNA ≤ 1 ×10⁷ copy/ml) and high viral load group (HBV DNA > 1 ×10⁷ copy/ml) with 95 cases in each group. Serum HBV DNA was detected by fluorescence real time quantitative PCR (RT-qPCR). In addition, serum samples from 95 healthy outpatients were collected simultaneously as the control group. The levels of mir-122 and mir-146a in serum exosomes were detected by RT-qPCR; Serum indexes of liver fibrosis were measured by chemiluminescence method. Pearson correlation analysis was applied for analyzing the association between serum exosome levels of mir-122 and mir-146a with HBV DNA load, and liver fibrosis indexes. Results Compared with the control group, the HBV DNA load, mir-122 level in serum exosomes, and the levels of serum hepatic fibrosis indexes including Hyaluronidase (HA), procollagen Ⅲ (PC Ⅲ), collagen Ⅳ(C Ⅳ) and laminin (LN) in low, medium and high HBV DNA load groups increased, whereas the level of mir-146a decreased in turn (P<0.05). The levels of mir-122 and mir-146a in serum exosome were found negatively correlated with HBV DNA load (r=0.765, P<0.05). The level of mir-122 was positively correlated with HBV DNA load and serum HA, PC Ⅲ, C Ⅳ and LN levels (r= 0.773, 0.764, 0.699, 0.665, 0.697, respectively, P<0.05), whereas the level of mir-146a was negatively correlated with these parameters (r=-0.840, -0.779, -0.731, -0.691, -0.738, respectively, P<0.05). Conclusion The expression and secretion of Mir-122 in serum exosome of CHB patients is up-regulated and that of mir-146a is down-regulated. They are closely co-related with HBV DNA load and may impact on HBV DNA replication, the severity of hepatic fibrosis, and the progression of CHB.

Key words: Chronic hepatitis B, microRNA-122, microRNA-146a, Hepatitis B virus, Exosome

刘娇, 刘青, 王大明, 贾兴旺. 慢性乙型肝炎患者血清外泌体mir-122、mir-146a表达与HBV DNA载量的相关性[J]. 肝脏, 2020, 25(10): 1038-1042.

LIU Jiao, LIU Qing, WANG Da-ming, JIA Xing-wang. The association of mir-122 and mir-146a levels in serum exosomes with HBV DNA load in chronic hepatitis B patients[J]. Chinese Hepatolgy, 2020, 25(10): 1038-1042.

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