Center for Gastroenterology and Hepatology, Institute of Liver Diseases, The Affiliated Hospital of Jiaxing College, Jiaxing, Zhejiang Province, PR China.
Department of Basic Medicine Sciences, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, PR China.
Commun Biol. 2021 Feb 18;4(1):230. doi: 10.1038/s42003-021-01747-5.
Wnt signaling dysfunction and gut dysbiosis may lead to liver fibrosis, yet the underlying mechanisms are not well elucidated. This study demonstrated the role of RSPO4, a Wnt signaling agonist, in liver fibrogenesis and its impact on the gut microbiome. RSPO4 gene in CCl-induced fibrotic-liver rats was knockout by Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) system, with healthy rats served as the control. Tissue samples and hepatic stellate cells (HSCs) isolated from rats were examined for curative effect of RSPO4-CRISPR treatment. Fecal sample were collected and analyzed with 16 S rRNA sequencing. We found RSPO4-CRISPR relieved liver fibrosis in rats and reversed HSC activation. Further, results showed RSPO4-CRISPR tended to restore the microflora composition. Significance species between groups were identified. Bacteroides and Escherichia-Shigella were the key microbes in the model and negative group, whereas Lactobacillus, Romboutsia, and Lachnospiraceae NK4A136 group were abundant in the control. Notably, Bacteroidales S24-7 group and Ruminococcaceae UCG-005 were the significantly enriched in CRISPR group. We show that the microbiome of rats treated with RSPO4-CRISPR presents a trend towards the restoration of the original condition. Our findings pave a new way to evaluate the curative effect of liver fibrosis treatment.
Wnt 信号功能障碍和肠道菌群失调可能导致肝纤维化，但潜在机制尚不清楚。本研究表明 Wnt 信号激动剂 RSPO4 在肝纤维化发生中的作用及其对肠道微生物组的影响。采用 Clustered Regularly Interspaced Short Palindromic Repeats（CRISPR）系统敲除 CCl4 诱导的纤维化肝脏大鼠的 RSPO4 基因，健康大鼠作为对照。检测 RSPO4-CRISPR 治疗对大鼠的疗效，对组织样本和分离的肝星状细胞（HSCs）进行检查。收集粪便样本并进行 16S rRNA 测序分析。我们发现 RSPO4-CRISPR 可缓解大鼠肝纤维化并逆转 HSC 激活。此外，结果表明 RSPO4-CRISPR 倾向于恢复微生物群落组成。鉴定了组间的重要物种。拟杆菌属和埃希氏菌-志贺氏菌属是模型和阴性组中的关键微生物，而乳杆菌属、罗姆布氏菌属和lachnospiraceae NK4A136 属在对照组中丰富。值得注意的是，Bacteroidales S24-7 组和 Ruminococcaceae UCG-005 在 CRISPR 组中明显富集。我们表明，用 RSPO4-CRISPR 治疗的大鼠的微生物组呈现出恢复原始状态的趋势。我们的发现为评估肝纤维化治疗的疗效开辟了新途径。
