| 胡毅翔,左清平,刘任祝,等.微 RNA-223-3p调控 NOD样受体热蛋白结构域 3/胱天氨酸蛋白酶 -1/白细胞介素 -1β信号通路抗小鼠肝纤维化的作用及机制[J].安徽医药,2023,27(8):1556-1561. |
| 微 RNA-223-3p调控 NOD样受体热蛋白结构域 3/胱天氨酸蛋白酶 -1/白细胞介素 -1β信号通路抗小鼠肝纤维化的作用及机制 |
| miR-223-3p exerts anti-mouse liver fibrosis effects through inhibition of NLRP3/Caspase-1/ IL-1β pathway |
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| DOI：10.3969/j.issn.1009-6469.2023.08.015 |
| 中文关键词: 肝纤维化 微 RNA-223-3p NOD样受体热蛋白结构域 3/胱天氨酸蛋白酶 -1/白细胞介素 -1β信号通路 炎症 前胶原 小鼠，近交 ICR |
| 英文关键词: Liver fibrosis miR-223-3p NLRP3/Caspase-1/IL-1β pathway Inflammation Procollagen Mice,inbred ICR |
| 基金项目:国家自然科学基金( 21877029)；湖南省自然科学基金( 2021JJ40549)；湖南省卫健委科研项目( 20201046) |
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| 中文摘要: |
| 目的探讨微 RNA-223-3p(miR-223-3p)调控 NOD样受体热蛋白结构域 3(NLRP3)/胱天氨酸蛋白酶 -1(Caspase-1)/白细胞介素 -1β(IL-1β)信号通路抗小鼠肝纤维化的作用及其机制。方法将 32只 ICR小鼠采用随机数字表法分为空白组、模型组、 miR-223-3p模拟物( miR-223-3p agomir)组和阴性对照 miRNA模拟物( agomir-NC)组，建立四氯化碳小鼠肝纤维化模型。 miR-223-3p agomir组和 agomir-NC组于第 4周末尾静脉注射 miR-223-3p agomir及 agomir-NC(给药剂量为每只 200 nmol/L)，每周给药 3次，连续给药 2周。分别比较 4组小鼠血清总胆红素( TBIL)、白蛋白( Alb)、血清透明质酸( HA)、 Ⅲ型前胶原( PCⅢ)、 Ⅳ型胶原( Ⅳ-C)表达水平，采用苏木精 -伊红( HE)染色及马松( Masson)染色检测肝组织病理改变。采用基因本体分析( GO)、 KEGG分析对 miR-223-3p进行功能分析，采用 Targetscan数据库对 miR-223-3p的靶基因进行预测，实时荧光定量 PCR(qPCR)及蛋白质印迹法( Western blotting)检测肝组织 NLRP3、Caspase-1、IL-1β mRNA及蛋白表达情况。结果与模型组相比， miR223-3p agomir组血清 TBIL［(24.32± 3.24)比( 62.21±4.52)］、 Alb［(34.12±7.32)比( 59.21±8.21)］、 HA［(136.12±10.25)比( 298.56±32.14)］、PCⅢ［(39.52±4.25)比( 78.32±14.21)］、Ⅳ-C［(50.25±6.32)比(165.85±12.35)］表达均明显降低( P<0.05)小鼠肝组织损伤得到明显改善，浸润的炎性细胞明显减少，纤维化程度明显减轻。 GO、KEGG富集及 miRNA-mRNA-KEGG关联，分析结果显示， miR-223-3p可在炎症、免疫等的 59个生命进程中发挥作用。靶点预测结果显示 NLRP3可能为 miR-223-3p的反向靶点。与模型组相比， miR-223-3p agomir组小鼠肝组织中 NLRP3、Caspase-1、IL-1β mRNA表达［( 1.24±0.12)比( 2.17±0.14)、(1.44±0.11)(2.65 ±0.18)、(1.31±0.13)比( 1.97±0.21)］及蛋白表达［(0.89± 0.05)比( 1.93 ±0.04)、(1.29± 0.07)比( 2.15 ±0.09)、(1.50± 0.05)2.52±0.12)］水平均显著降低( P<0.05)。结论过表达 miR-223-3p可抑制 NLRP3/Caspase-1/IL-1β信号通路的激活，降低机 比(比体炎症反应，改善小鼠肝组织损伤，进而发挥其抗肝纤维化作用。 |
| 英文摘要: |
| Objective To investigate the effect and mechanism of microRNA-223-3p (miR-223-3p) on anti-hepatic fibrosis in mice by regulating NOD-like receptor pyridine domain containing 3 (NLRP3)/caspase-1/interleukin-1β (IL-1β) signaling pathway. Meth ods Thirty-two ICR mice were randomly divided into blank group, model group, miR-223-3p mimic (miR-223-3p agomir) agomir group and negative control miRNA mimic (agomir-NC) group, and a carbon tetrachloride mouse model of liver fibrosis was established. miR-223-3p agomir group and agomir-NC group were injected with miR-223-3p agomir and agomir-NC (200 nM per mouse) via tailvein at the end of the 4th week, 3 times a week for 2 weeks. The expression levels of serum total bilirubin (TBIL), albumin (Alb), serumhyaluronic acid (HA), type Ⅲ procollagen (PCⅢ), and type Ⅳ collagen (Ⅳ -C) were compared among the four groups. Pathological changes in liver tissue were observed by hematoxylin-eosin (HE)staining and Masson staining. GO and KEGG analyses were used to an alyze miR-223-3p function. Targetscan database was used to predict miR-223-3p target genes. The mRNA and protein levels of NL RP3, Caspase-1, and IL-1β in hepatic tissue were determined by real-time fluorescent quantitative PCR (q-PCR) and western blotting. Results Compared with the model group, serum TBIL [(24.32± 3.24) vs. (62.21 ±4.52)], Alb [(34.12±7.32) vs. (59.21±8.21)], HA [(136.12±10.25) vs. (298.56 ±32.14) ], PCⅢ [(39.52±4.25) vs. (78.32±14.21)], Ⅳ-eob-C [(50.25±6.32) vs. (165.85±12.35)] in miR-2233p agomir group were significantly decreased (P<0.05). The liver tissue injury of mice was significantly improved, the infiltration of inflammatory cells was significantly reduced, and the degree of fibrosis was significantly reduced. The GO, KEGG enrichment results,and miRNA-mRNA-KEGG correlation analysis showed that miR-223-3p could play a role in 59 life processes, such as inflammationand immunity. Target prediction results showed that NLRP3 might be the downstream target of miR-223-3p. Compared with the model group, The mRNA levels of NLRP3, Caspase-1, and IL-1β in miR-223-3p agomir group [(1.24±0.12) vs. (2.17±0.14), (1.44±0.11) vs. (2.65±0.18), (1.31±0.13) vs. (1.97±0.21)] and protein expression [(0.89±0.05) vs. (1.93±0.04),(1.29±0.07) vs. (2.15 ±0.09),(1.50±0.05) vs. (2.52±0.12)] were significantly decreased (P<0.05).Conclusion Overexpression of miR-223-3p inhibits the activation of NLRP3/ Caspase-1/IL-1β signaling pathway, reduces the inflammatory response and improves liver tissue damage in mice, thus exerting its anti-liver fibrosis effect |
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