Integrating serum exosomal microRNA and liver microRNA profiles disclose the function role of autophagy and mechanisms of Fructus Meliae Toosendan-induced hepatotoxicity in mice
Integrating serum exosomal microRNA and liver microRNA profiles disclose the function role of autophagy and mechanisms of Fructus Meliae Toosendan-induced hepatotoxicity in mice
复制标题
整合血清外泌体 microRNA 和肝脏 microRNA 图谱揭示自噬的功能作用以及苦楝子诱导小鼠肝毒性的机制
DOI：
10.1016/j.biopha.2019.109709
复制
发表时间：
2020
影响因子：
7.5
通讯作者：
Xiaohui Fan
中科院分区：
文献类型：
--
作者：
Lingqi Yu;Jie Zheng;Junying Li;Yuzhen Wang;Xiaoyan Lu;Xiaohui Fan
草药性肝损伤（HILI）是一个日益严重的临床和经济问题。然而，HILI的潜在机制在很大程度上仍然未知，这阻碍了这种疾病的预防和治疗。最近的证据支持循环外泌体和细胞中的microRNAs （miRNAs）在肝脏疾病的病理中起重要作用。因此，本研究旨在通过对FMT乙酸乙酯提取物（FMT）暴露小鼠血清外泌体miRNAs和肝脏miRNAs谱的综合分析，揭示FMT诱导肝损伤（FILI）的机制。本研究采用20和40 g/kg两种剂量的FMT，仅高剂量暴露对小鼠造成明显的肝损伤。对高剂量FMT与对照组血清外泌体中209个差异表达mirna （DEMs）的通路分析显示，FILI可能受到凋亡相关通路的调控，如p53信号、PI3K/Akt信号和PTEN信号。综合分析高剂量FMT组血清外泌体dem和肝脏dem的mRNA靶点，发现自噬显著富集，是FILI的顶级典型途径之一。TUNEL法证实高剂量fmt处理小鼠肝组织中存在肝细胞凋亡。此外，体内验证研究表明，大剂量FMT给药后，小鼠肝脏中PTEN、p-AKT、p53和BAX的蛋白表达水平确实受到调节，表明肝细胞凋亡可能通过上述三种途径介导。有趣的是，PINK1/ parkin介导的线粒体自噬在高剂量fmt处理的小鼠肝脏中被激活，自噬对FILI的保护作用在体外通过自噬通量抑制剂得到验证。此外，血清外泌体miR-222是低剂量和高剂量FMT治疗中下调最多的mirna，可能通过调节PTEN和PPP2R2A在肝细胞凋亡中起重要作用。综上所述，综合分析小鼠血清外泌体和肝细胞的microRNA谱，可以深入了解FMT的肝毒性机制，揭示自噬在FILI中的保护作用，表明该方法有助于深入了解HILI的机制，激活自噬可能是治疗FILI甚至HILI的潜在策略。
Herb-induced liver injury (HILI) is a growing clinical and economic problem worldwide. However, the underlying mechanism of HILI remains largely unknown, which hinders the prevention and treatment of this disease. Recently evidence supports that microRNAs (miRNAs) in circulating exosomes and cells play an important role in the pathology of liver diseases. Thus, using Fructus Meliae Toosendan (FMT) as an example of hepatoxic herbal medicine, the aim of this study was to reveal the mechanisms of FMT-induced liver injury (FILI) through integrated analysis of serum exosomal miRNAs and liver miRNAs profiles on FMT ethyl acetate extract (FMT for short)-exposed mice. Two dosages of FMT (20 and 40 g/kg) were involved in this study, while only high-dose exposure induced obvious liver injury in mice. Pathway analysis of 209 differentially expressed miRNAs (DEMs) in serum exosomes between high-dose FMT and control groups exhibited that FILI might be regulated by apoptosis-related pathways, such as p53 signaling, PI3K/Akt signaling, and PTEN signaling. Integrated analysis of the mRNA targets of serum exosomal DEMs and liver DEMs of high-dose FMT group showed that autophagy was significantly enriched as one of the top canonical pathways in FILI. Hepatocyte apoptosis was then proved by TUNEL assay in the liver tissue of high-dose FMT-treated mice. Moreover,in vivovalidation studies suggested that the protein expression levels of PTEN, p-AKT, p53, and BAX were indeed regulated in the mouse liver after high-dose FMT administration, indicating hepatocyte apoptosis may be mediated by these three pathways mentioned above. Intriguingly, PINK1/Parkin-mediated mitophagy was activated in high-dose FMT-treated mouse liver and the protective effect of autophagy in FILI was validatedin vitrowith an autophagic flux inhibitor. In addition, serum exosomal miR-222, the most downregulated miRNAs between low- and high-dose FMT treatments, might be an important event in the hepatocyte apoptosis by regulating PTEN and PPP2R2A. In conclusion, integrated analysis of microRNA profiles in mouse serum exosomes and liver cells provides insights into the hepatotoxicity mechanisms of FMT and discloses the protective role of autophagy in FILI, suggesting this method could contribute to deeply understand the mechanism of HILI and activation of autophagy may be a potentially therapeutic strategy for FILI even HILI.