Broermann Andre, Schmid Ramona, Gabrielyan Ogsen, Sakowski Marlene, Eisele Claudia, Keller Sascha, Wolff Michael, Baum Patrick, Stierstorfer Birgit, Huber Jochen, Krämer Bernhard K, Hocher Berthold, Streicher Ruediger, Delić Denis
Cardiometabolic Diseases Research, Boehringer Ingelheim Pharma GmbH & Co. KG, Birkendorferstr.65, 88397 Biberach Germany.
Translational Medicine & Clinical Pharmacology, Boehringer Ingelheim Pharma GmbH & Co. KG, Birkendorferstr.65, 88397 Biberach, Germany.
Int J Mol Sci. 2020 Dec 31;22(1):382. doi: 10.3390/ijms22010382.
MicroRNAs (miRNAs) are short, non-coding RNA species that are important post-transcriptional regulators of gene expression and play an important role in the pathogenesis of non-alcoholic fatty liver disease. Here, we investigated the phosphodiesterase 5 (PDE5) inhibitor induced effects on hepatic and plasma exosomal miRNA expression in CCl-treated rats. In the present study, hepatic miRNA profiling was conducted using the Nanostring nCounter technology and mRNA profiling using RNA sequencing from PDE5 treated rats in the model of CCl-induced liver fibrosis. To evaluate if the PDE5 inhibitor affected differentially expressed miRNAs in the liver can be detected in plasma exosomes, qRT-PCR specific assays were used. In livers from CCl-treated rats, the expression of 22 miRNAs was significantly increased (> 1.5-fold, adj. < 0.05), whereas the expression of 16 miRNAs was significantly decreased (> 1.5-fold, adj. < 0.05). The majority of the deregulated miRNA species are implicated in fibrotic and inflammatory processes. The PDE5 inhibitor suppressed the induction of pro-fibrotic miRNAs, such as miR-99b miR-100 and miR-199a-5p, and restored levels of anti-fibrotic miR-122 and miR-192 in the liver. In plasma exosomes, we observed elevated levels of miR-99b, miR-100 and miR-142-3p after treatment with the PDE5-inhibitor compared to CCl/Vehicle-treated. Our study demonstrated for the first time that during the development of hepatic fibrosis in the preclinical model of CCl-induced liver fibrosis, defined aspects of miRNA regulated liver pathogenesis are influenced by PDE5 treatment. In conclusion, miRNA profiling of plasma exosomes might be used as a biomarker for NASH progression and monitoring of treatment effects.
微小RNA（miRNA）是短链非编码RNA，是基因表达重要的转录后调节因子，在非酒精性脂肪性肝病的发病机制中起重要作用。在此，我们研究了磷酸二酯酶5（PDE5）抑制剂对四氯化碳处理大鼠肝脏和血浆外泌体miRNA表达的影响。在本研究中，使用Nanostring nCounter技术进行肝脏miRNA谱分析，并使用RNA测序对四氯化碳诱导肝纤维化模型中经PDE5处理的大鼠进行mRNA谱分析。为了评估PDE5抑制剂对肝脏中差异表达的miRNA在血浆外泌体中是否可被检测到，采用了qRT-PCR特异性检测方法。在四氯化碳处理大鼠的肝脏中，22种miRNA的表达显著增加（>1.5倍，校正P<0.05），而16种miRNA的表达显著降低（>1.5倍，校正P<0.05）。大多数失调的miRNA与纤维化和炎症过程有关。PDE5抑制剂抑制了促纤维化miRNA如miR-99b、miR-100和miR-199a-5p的诱导，并恢复了肝脏中抗纤维化miR-122和miR-192的水平。在血浆外泌体中，与四氯化碳/溶剂处理组相比，用PDE5抑制剂处理后，我们观察到miR-99b、miR-100和miR-142-3p水平升高。我们的研究首次证明，在四氯化碳诱导肝纤维化的临床前模型中肝纤维化发展过程中，miRNA调节的肝脏发病机制的特定方面受PDE5治疗的影响。总之，血浆外泌体的miRNA谱分析可能用作非酒精性脂肪性肝炎进展和治疗效果监测的生物标志物。
