胆道闭锁患儿血清microRNA-21水平的表达及与肝纤维化的相关性分析

Expression of serum microRNA-21 in children with biliary atresia and its correlation with liver fibrosis

摘要:

目的 探讨胆道闭锁患儿血清microRNA-21（miR-21）水平的表达及与肝纤维化的相关性。方法 选取2017年7月—2019年7月河北省儿童医院收治的先天性胆道闭锁患儿76例作为胆道闭锁组，同期在该院进行体检的健康婴幼儿50例作为正常对照组。采用qRT-PCR法检测研究对象血清miR-21 mRNA相对表达量，进一步根据胆道闭锁组的miR-21 mRNA相对表达量中位数将其分为高miR-21组和低miR-21组，每组38例。比较不同miR-21 mRNA相对表达量胆道闭锁患儿的血清总胆红素水平、肝组织纤维化评分及天冬氨酸转氨酶与血小板比值（APRI）、血清肝纤维化指标［层粘连蛋白（LN）、透明质酸酶（HA）、Ⅲ型前胶原（PCⅢ）、Ⅳ型胶原（Ⅳ-Col）］水平、肝脏组织上皮-间质细胞转化（EMT）相关分子［E-cadherin、ZEB1］水平的差异。分析胆道闭锁患儿血清miR-21与肝纤维化的相关性。结果 高miR-21组、低miR-21组、正常对照组血清miR-21 mRNA相对表达量和总胆红素水平比较，差异有统计学意义（P <0.05），高miR-21组血清miR-21 mRNA相对表达量和总胆红素水平高于低miR-21组；低miR-21组miR-21 mRNA相对表达量和总胆红素水平高于正常对照组。高miR-21组肝组织纤维化评分、APRI、LN、HIA、PCⅢ、Ⅳ-Col水平高于低miR-21组（P <0.05）。高miR-21组肝脏组织E-cadherin mRNA相对表达量低于低miR-21组（P <0.05）；ZEB1 mRNA相对表达量高于低miR-21组（P <0.05）。Pearson相关分析发现，胆道闭锁患儿血清miR-21水平与血清总胆红素水平、肝组织纤维化评分、APRI呈正相关（r =0.546、0.669和0.578，均P <0.05）；与肝纤维化指标LN、HA、PCⅢ、Ⅳ-Col的水平呈正相关（r =0.781、0.663、0.503和0.628，均P <0.05）；与E-cadherin mRNA相对表达量呈负相关（r =-0.614，P <0.05），与ZEB1 mRNA相对表达量呈正相关（r =0.607，P <0.05）。结论 高表达的miR-21可能参与胆道闭锁患儿肝纤维化过程，可能与其促进EMT的作用相关。

Abstract:

Objective To investigate the expression of serum microRNA-21 in children with biliary atresia and its correlation with liver fibrosis.Methods From July 2017 to July 2019, 76 children with congenital biliary atresia admitted to our hospital were selected as the biliary atresia group, and 50 healthy children who underwent physical examination in our hospital during the same period were regarded as the normal control group. The expression level of serum microRNA-21 of the subjects were detected, and the subjects were further divided into high microRNA-21 group and low microRNA-21 group, 38 cases in each group, according to the median expression level of microRNA-21 in the biliary atresia group. The different microRNA-21 expression level of children with biliary atresia in serum total bilirubin, liver tissue fibrosis score, aspartate aminotransferase (APRI), platelet ratio value, serum liver fibrosis indexes [laminin (LN), hyaluronic acid (HA) enzyme, procollagen type Ⅲ (PCⅢ), collagen type Ⅳ (Ⅳ-Col)] level, epithelial stromal transformation (EMT) in liver tissue, related molecular [E-cadherin, ZEB1] expression level were compared. The correlation between serum microRNA-21 expression level and liver fibrosis in children with biliary atresia was analyzed.Results The expression level of microRNA-21 and serum total bilirubin level among high microRNA-21 group, low microRNA-21 group, and normal control group were compared by one-way ANOVA, the difference was statistically significant (P < 0.05). MicroRNA-21, serum total bilirubin, LN, HA, PCⅢ, Ⅳ-Col level, Liver fibrosis scores, APRI values, and the expression of ZEB1 mRNA in liver tissues in high microRNA-21 group were higher than those in low microRNA-21 group (P < 0.05); the difference in the degree of fibrosis between high microRNA-21 group and low microRNA-21 group was also statistically significant (P < 0.05); the expression of E-cadherin mRNA in liver tissues of high microRNA-21 group was lower than that of low microRNA-21 group (P < 0.05). Pearson correlation analysis found that expression level of serum microRNA-21 in children with biliary atresia was positively correlated with serum total bilirubin level (r = 0.546), liver fibrosis score (r = 0.669), and APRI value (r = 0.578) (P < 0.05), was positively correlated with levels of liver fibrosis indexes such as LN (r = 0.781), HA (r = 0.663), PCⅢ (r = 0.628), Ⅳ-Col (r = 0.503) (P < 0.05), was negatively correlated with expression of EMT related molecular E-cadherin (r = -0.614), and was positively correlated with expression of ZEB1 mRNA (r = 0.607) (all P < 0.05).Conclusion Highly expressed microRNA-21 is probably involved in the process of liver fibrosis in children with biliary atresia, which may be related to its role in promoting EMT.