Astragalus Ameliorates Peritoneal Fibrosis by Inhibiting Myostatin via miR-27a-3p

Background: Astragalus improved peritoneal fibrosis (PF) through several fibrotic signals. Myostatin mediated protein synthesis and catabolism were associated with tissue fibrosis. However, the proteolysiscould take part in PF, and the mechanism of by whichAstragalus exerts multi-target regulation remains unknown. This study will illustrate the molecular mechanism that Astragalus improve PF through Myostatin andproteolysis, on the basis of crosstalk of peritoneum and skeletal muscle. Methods: Firstly, we observed muscle consumption, Myostatin, markers of proteolysis in PF and Astragalus-treated model. Secondly, Myostatin siRNA was used to explore whether Myostatin mediated proteolysis was involved in PF and effect of Astragalus. Thirdly, mRNA sequencingand enrichment analysis were screen for differential mRNA and biological process. Network pharmacology was used to confirm the common target for PF and muscle consumption, which could regulate by Astragalus. Fourthly, miRNA sequencing, TargetScan and luciferase reporter were screen for differential miRNA, which could bind with Myostatin 3’-UTR. Fifthly, we used Agomir to label miR-27a-3p. miR-27a-3p mimic was injected into the anterior tibial muscle to explore the molecular mechanism of Astragalus improving PF by miR-27a-3p binding Myostatin. Lastly, markers of PF, Myostatin and miR-27a-3p were detected from peritoneal dialysis effluent. Results: 1. Astragalus inhibited Myostatin, FBXO32 and TRIM63 expression, which act on improving PF and muscle consumption. 2. Myostatin and proteolysiswere involved in pathological processes of PF. 3. mRNA sequencing and enrichment analysis showed that protein metabolism in muscle cells is associated with PF, and Myostatin is a common target of PF and skeletal muscle consumption regulated by Astragalus. 4. miRNA sequencing and luciferase reporter assays were confirmed that miR-27a-3p can target Myostatin 3’-UTR and inhibit Myostatin mRNA. Astragalus enhanced the expression of miR-27a-3p. 5. Astragalus improved PF by miR-27a-3p targeting Myostatin mRNA, the mechanism was associated with Myostatin mediated proteolysis. 6. The expression of Myostatin, miR-27a-3p and markers of fibrosis from peritoneal dialysis effluent were increased within longer peritoneal dialysis. Conclusion: Myostatin mediated proteolysis plays an important role in PF. Astragalus improved PF by miR-27a-3p targeting Myostatin mRNA. Funding: Government Support – Non-U.S.