[Effect of Isodon ternifolius-medicated serum on hepatic stellate cells based on TLR4/NF-κB/NLRP3 signaling pathway]

Zhongguo Zhong Yao Za Zhi. 2023 Jul;48(14):3913-3921. doi: 10.19540/j.cnki.cjcmm.20230306.704.
[Article in Chinese]

Abstract

The present study aimed to investigate the inhibitory effect and mechanism of Isodon terricolous-medicated serum on lipopolysaccharide(LPS)-induced hepatic stellate cell(HSC) activation. LPS-induced HSCs were divided into a blank control group, an LPS model group, a colchicine-medicated serum group, an LPS + blank serum group, an I. terricolous-medicated serum group, a Toll-like receptor 4(TLR4) blocker group, and a TLR4 blocker + I. terricolous-medicated serum group. HSC proliferation was detected by methyl thiazolyl tetrazolium(MTT) assay. Enzyme-linked immunosorbent assay(ELISA) was used to measure type Ⅰ collagen(COL Ⅰ), COL Ⅲ, transforming growth factor-β1(TGF-β1), intercellular adhesion molecule-1(ICAM-1), α-smooth muscle actin(α-SMA), vascular cell adhesion molecule-1(VCAM-1), cysteinyl aspartate-specific proteinase-1(caspase-1), and monocyte chemotactic protein-1(MCP-1). Real-time PCR(RT-PCR) was used to detect mRNA expression of TLR4, IκBα, and NOD-like receptor thermal protein domain associated protein 3(NLRP3), nuclear factor-κB(NF-κB) p65, gasdermin D(GSDMD), and apoptosis-associated speck-like protein containing a CARD(ASC) in HSCs. Western blot(WB) was used to detect the protein levels of TLR4, p-IκBα, NF-κB p65, NLRP3, ASC, and GSDMD in HSCs. The results showed that I. terricolous-medicated serum could inhibit the proliferation activity of HSCs and inhibit the secretion of COL Ⅰ, COL Ⅲ, α-SMA, TGF-β1, caspase-1, MCP-1, VCAM-1, and ICAM-1 in HSCs. Compared with the LPS model group, the I. terricolous-medicated serum group, the colchicine-medicated serum group, and the TLR4 blocker group showed down-regulated expression of p-IκBα, NLRP3, NF-κB p65, GSDMD, and ASC, and up-regulated expression of IκBα. Compared with the TLR4 blocker group, the TLR4 blocker + I. terricolous-medicated serum group showed decreased expression of TLR4, p-IκBα, NLRP3, NF-κB p65, GSDMD, and ASC, and increased expression of IκBα. In conclusion, I. terricolous-medicated serum down-regulates HSC activation by inhibiting the TLR4/NF-κB/NLRP3 signaling pathway.

Keywords: Isodon ternifolius; TLR4/NF-κB/NLRP3; hepatic stellate cells.

Publication types

  • English Abstract

MeSH terms

  • Caspases
  • Colchicine / metabolism
  • Colchicine / pharmacology
  • Hepatic Stellate Cells
  • Intercellular Adhesion Molecule-1 / metabolism
  • Isodon*
  • Lipopolysaccharides / pharmacology
  • NF-KappaB Inhibitor alpha / metabolism
  • NF-kappa B* / genetics
  • NF-kappa B* / metabolism
  • NLR Family, Pyrin Domain-Containing 3 Protein / genetics
  • NLR Family, Pyrin Domain-Containing 3 Protein / metabolism
  • Signal Transduction
  • Toll-Like Receptor 4 / genetics
  • Toll-Like Receptor 4 / metabolism
  • Transforming Growth Factor beta1 / metabolism
  • Vascular Cell Adhesion Molecule-1 / metabolism

Substances

  • NF-kappa B
  • Transforming Growth Factor beta1
  • NF-KappaB Inhibitor alpha
  • Intercellular Adhesion Molecule-1
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • Toll-Like Receptor 4
  • Vascular Cell Adhesion Molecule-1
  • Lipopolysaccharides
  • Colchicine
  • Caspases