Isoliensinine Attenuates Renal Fibrosis and Inhibits TGF-β1/Smad2/3 Signaling Pathway in Spontaneously Hypertensive Rats

Drug Des Devel Ther. 2023 Sep 7:17:2749-2762. doi: 10.2147/DDDT.S414179. eCollection 2023.

Abstract

Purpose: This study aimed to investigate the molecular mechanisms of isoliensinine, a kind of bibenzyl isoquinoline alkaloid which isolated from a TCM named Lotus Plumule (Nelumbo nucifera Gaertn), in treating renal interstitial fibrosis (RIF) by using RNA sequencing, KEGG analysis and in vivo experimental approaches.

Methods: Spontaneous hypertension rats (SHRs) were randomly assigned into five groups, consisting of SHR, SHR+Isoliensinine-L (2.5 mg/kg/day), SHR+Isoliensinine-M (5 mg/kg/day), SHR+Isoliensinine-H (10 mg/kg/day), and SHR+Valsartan (10 mg/kg/day) groups (n = 6 for each group). A control group of Wistar Kyoto rats (n = 6) was also included. Rats were treated intragastrically with isoliensinine, valsartan, or double-distilled water of equal volume for 10 weeks. To examine the therapeutic impact on hypertensive renal injury, fibrosis, and its underlying mechanisms, multiple techniques were employed, including hematoxylin and eosin staining, Masson trichrome staining, RNA sequencing, gene ontology (GO) function and pathway enrichment analysis and immunohistochemistry.

Results: Resultantly, the use of isoliensinine at different concentrations or valsartan showed significant improvement in renal pathological injury in SHRs. RNA sequencing and KEGG analysis uncovered 583 differentially expressed transcripts and pathways enriched in collagen formation and ECM-receptor interaction after treatment with isoliensinine. There was also a reduction in the increase of collagen and upregulation of collagen I & III, TGF-β1, p-Smad2, and p-Smad3 in the renal tissue of SHRs. Thus, isoliensinine ameliorated renal injury and collagen deposition in hypertensive rats, and inhibiting the activation of the TGF-β1/Smad2/3 pathway might be one of the underlying mechanisms.

Conclusion: This study showed that treatment with isoliensinine effectively reduced the renal injury and fibrosis in SHRs. In addition, isoliensinine inhibited the TGF-β1/Smad2/3 signaling in-vivo. These findings provided strong evidence for the therapeutic benefits of isoliensinine in combating renal injury and fibrosis.

Keywords: RNA sequencing; TGF-β1/Smad2/3 pathway; collagen deposition; hypertension; isoliensinine; renal injury.

MeSH terms

  • Animals
  • Fibrosis
  • Isoquinolines / pharmacology
  • Kidney Diseases* / drug therapy
  • Rats
  • Rats, Inbred SHR
  • Signal Transduction
  • Transforming Growth Factor beta1*

Substances

  • isoliensinine
  • Transforming Growth Factor beta1
  • Isoquinolines

Grants and funding

This study was sponsored by the National Natural Science Foundation of China (82074363), the National Natural Science Foundation of China (82204662), the National Natural Science Foundation of China (U22A20372) the Science and Technology Major Project of Fujian Province (2019YZ014004), the Young Elite Scientists Sponsorship Program of the China Association of Chinese Medicine (2021-QNRC2-B19), Scientific and economic integration service platform for translational medicine of cardiovascular diseases in Fujian Province, Fuzhou (2021XRH004); the Development Fund of Chen Keji Integrative Medicine (CKJ2020003).