Inflammation and Immune Mediated Diseases Laboratory of Anhui Province, Anhui Institute of Innovative Drugs, School of Pharmacy, Anhui Medical University, Hefei 230032, China; The KeyLaboratory of Anti-inflammatory and Immune Medicines, Anhui Medical University, Ministry of Education, Hefei 230032, China; Institute for Liver Diseases of Anhui Medical University, ILD-AMU, Anhui Medical University, Hefei 230032, China.
Inflammation and Immune Mediated Diseases Laboratory of Anhui Province, Anhui Institute of Innovative Drugs, School of Pharmacy, Anhui Medical University, Hefei 230032, China; The KeyLaboratory of Anti-inflammatory and Immune Medicines, Anhui Medical University, Ministry of Education, Hefei 230032, China.
Int Immunopharmacol. 2022 Sep;110:109034. doi: 10.1016/j.intimp.2022.109034. Epub 2022 Jul 11.
Hepatic fibrosis is an essential pathology of multiple chronicliverdiseases. The aim of this study was to investigate the role of miR-301a-3p in hepatic fibrosis. We found that miR-301a-3p was upregulated in hepatic fibrosis patients and in culture-activated human hepatic stellate cells (HSCs). Interestingly, miR-301a-3p expression was increased in hepatic fibrosis progression mice while decreased in hepatic fibrosis recovery mice, indicating that miR-301a-3p may participate in the hepatic fibrosis pathology. Functionally, the effects of miR-301a-3p both on hepatic fibrosis progression and regression were assessed in vivo. Inhibiting miR-301a-3p amelioratedmouse liver fibrogenesis and collagen deposition and suppressed HSC activation and fibrogenic factor expression. Whereas, in hepatic fibrosis regression, upregulating miR-301a-3p impaired mouse hepatic fibrosis recovery by inducing HSC activation and triggering inflammation. Consistently, gain-of-function and loss-of-function analysis of miR-301a-3p were performed to evaluate its effects on human HSCs LX-2 cell. We found that suppressing miR-301a-3p inhibited LX-2 cell activation and proliferation, and induced LX-2 cell apoptosis, accompaniedby decreased fibrotic mediators expression. Collectively, these findings suggest miR-301a-3p drives liver fibrogenesis and HSC activation in hepatic fibrosis. Mechanistically, we demonstrated miR-301a-3p binds directly to phosphatase and tensin homolog (PTEN) by luciferase reporter analysis, pull-down, and RIP assay. Indicating that miR-301a-3p plays a critical rolein promotingliverfibrogenesis viamodulating the PTEN/platelet derived growth factor β (PDGFR-β) pathway. In conclusion, our findings demonstrate that miR-301a-3p expression is closely correlated with hepatic fibrosis pathology, and that enhancing miR-301a-3p maintains the HSC profibrogenic phenotype, triggers inflammatoryresponses, promotes fibrogenic factor production, and further exacerbates liver fibrogenesis. These findings suggest that miR-301a-3p may serve as a promising diagnostic and prognosis biomarker for hepatic fibrosis treatment.
肝纤维化是多种慢性肝病的重要病理。本研究旨在探讨 miR-301a-3p 在肝纤维化中的作用。我们发现 miR-301a-3p 在肝纤维化患者和培养激活的人肝星状细胞（HSCs）中上调。有趣的是，miR-301a-3p 的表达在肝纤维化进展小鼠中增加，而在肝纤维化恢复小鼠中减少，表明 miR-301a-3p 可能参与肝纤维化病理。功能上，在体内评估了 miR-301a-3p 对肝纤维化进展和恢复的影响。抑制 miR-301a-3p 可改善小鼠肝纤维化和胶原沉积，并抑制 HSC 激活和纤维化因子表达。然而，在肝纤维化恢复中，上调 miR-301a-3p 通过诱导 HSC 激活和触发炎症来损害小鼠肝纤维化恢复。一致地，进行了 miR-301a-3p 的功能增益和功能丧失分析，以评估其对人 HSCs LX-2 细胞的影响。我们发现，抑制 miR-301a-3p 抑制了 LX-2 细胞的激活和增殖，并诱导了 LX-2 细胞凋亡，同时伴有纤维化介质表达的降低。总之，这些发现表明 miR-301a-3p 在肝纤维化中驱动肝纤维化和 HSC 激活。机制上，我们通过荧光素酶报告分析、下拉和 RIP 测定证实 miR-301a-3p 直接与磷酸酶和张力蛋白同源物（PTEN）结合。表明 miR-301a-3p 通过调节 PTEN/血小板衍生生长因子β（PDGFR-β）通路在促进肝纤维化中发挥关键作用。总之，我们的发现表明 miR-301a-3p 的表达与肝纤维化病理密切相关，增强 miR-301a-3p 维持 HSC 致纤维化表型，引发炎症反应，促进纤维化因子产生，并进一步加重肝纤维化。这些发现表明 miR-301a-3p 可能作为肝纤维化治疗的有前途的诊断和预后生物标志物。
