Fasudil alleviated insulin resistance through promotion of proliferation, attenuation of cell apoptosis and inflammation and regulation of RhoA/Rho kinase/insulin/nuclear factor-κB signalling pathway in HTR-8/SVneo cells

J Pharm Pharmacol. 2021 Jul 7;73(8):1118-1127. doi: 10.1093/jpp/rgab033.

Abstract

Objectives: The aim of this study was to evaluate the effects of fasudil on insulin resistance (IR) in HTR-8/SVneo cells.

Methods: HTR-8/SVneo cells were treated with insulin or/and fasudil. Cell proliferation, apoptosis, inflammation and related signalling pathways were assessed.

Key findings: Insulin treatment significantly enhanced the protein expressions of RhoA and Rho kinase (ROCK1 and ROCK2), but decreased glucose consumption. Administration of fasudil effectively promoted glucose uptake. Moreover, fasudil enhanced cell viability and the level of proliferating cell nuclear antigen (PCNA). Insulin-mediated cell apoptosis was inhibited by fasudil via the down-regulation of bax and cleaved-caspase-3, and the up-regulation of bcl-2. At the same time, fasudil led to the reduction of IL-1β, TNF-α, IL-6 and IL-8 mRNA levels in insulin-treated cells. In addition, RhoA, ROCK2 and phosphorylated myosin phosphatase target subunit-1 (p-MYPT-1) expressions were down-regulated by fasudil. Importantly, fasudil activated insulin receptor substrate-1 (IRS-1) through increasing p-IRS-1 (Tyr612) and p-Akt expressions. The nuclear NF-κB p65 and p-IκB-α levels were reduced via the administration of fasudil in insulin-treated cells.

Conclusions: Fasudil mitigated IR by the promotion of cell proliferation, inhibition of apoptosis and inflammation and regulation of RhoA/ROCK/insulin/NF-κB signalling pathway through in vitro studies.

Keywords: HTR-8/SVneo cells; RhoA/Rho kinase/insulin/NF-κB signalling pathway; fasudil; insulin resistance.

MeSH terms

  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / analogs & derivatives*
  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / pharmacology
  • Apoptosis / drug effects
  • Caspase 3 / metabolism
  • Cell Survival
  • Cells, Cultured
  • Diabetes, Gestational / drug therapy
  • Diabetes, Gestational / metabolism
  • Female
  • Humans
  • Insulin Resistance*
  • Insulin* / metabolism
  • Insulin* / pharmacology
  • NF-kappa B / metabolism
  • Pregnancy
  • Proliferating Cell Nuclear Antigen / metabolism
  • Protein Kinase Inhibitors / pharmacology
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Signal Transduction / drug effects*
  • Treatment Outcome
  • Trophoblasts* / metabolism
  • Trophoblasts* / pathology
  • rho-Associated Kinases* / antagonists & inhibitors
  • rho-Associated Kinases* / metabolism

Substances

  • BCL2 protein, human
  • Insulin
  • NF-kappa B
  • PCNA protein, human
  • Proliferating Cell Nuclear Antigen
  • Protein Kinase Inhibitors
  • Proto-Oncogene Proteins c-bcl-2
  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
  • rho-Associated Kinases
  • CASP3 protein, human
  • Caspase 3
  • fasudil