Atorvastatin Combined with Low-Dose Dexamethasone Treatment Protects Endothelial Function Impaired by Chronic Subdural Hematoma via the Transcription Factor KLF-2

Drug Des Devel Ther. 2020 Aug 12:14:3291-3299. doi: 10.2147/DDDT.S256050. eCollection 2020.

Abstract

Objective: Our previous study showed that the combination therapy with atorvastatin and low-dose dexamethasone protected endothelial cell function in chronic subdural hematoma (CSDH) injury. In this study, we aimed to investigate the mechanism underlying the effects of this combination therapy on CSDH-induced cell dysfunction.

Methods: Monocytes and endothelial cells were cocultured with CSDH patient hematoma samples to mimic the pathological microenvironment of CSDH. Monocytes (THP-1 cells) and endothelial cells (hCMEC/D3 cells) were cocultured in a transwell system for 24 h before stimulation with hematoma samples diluted in endothelial cell medium (ECM) at a 1:1 ratio. Tight junction markers were detected by Western blotting, PCR and immunofluorescence. hCMEC/D3 cells were collected for Western blot and PCR analyses to detect changes in the expression levels of vascular cell adhesion molecule (VCAM-1), intercellular adhesion molecule (ICAM-1), and Kruppel-like factor 2 (KLF-2). The IL-6, IL-10 and VEGF levels in the supernatant were measured by enzyme-linked immunosorbent assay (ELISA).

Results: KLF-2 expression in endothelial cells was decreased after stimulation with CSDH patient hematoma samples, but combination therapy with atorvastatin and low-dose dexamethasone reversed this trend. KLF-2 protected injured cells by increasing the expression of VE-cadherin and ZO-1; attenuating the expression of VCAM-1, ICAM-1, IL-6 and VEGF; and enhancing the expression of IL-10, all of which play pivotal roles in endothelial inflammation. Moreover, the effect of combination therapy with atorvastatin and low-dose dexamethasone was obviously reduced in endothelial cells with KLF-2 knockdown compared with normal cells.

Conclusion: Coculture with hematoma samples decreased KLF-2 expression in human cerebral endothelial cells. Combination therapy with atorvastatin and low-dose dexamethasone counteracted hematoma-induced KLF-2 suppression in human cerebral endothelial cells to attenuate robust endothelial inflammation and permeability. KLF-2 plays an important role in drug therapy for CSDH and may become the key factor in treatment and prognosis.

Keywords: Kruppel-like factor 2; atorvastatin combined with low-dose dexamethasone treatment; chronic subdural hematoma; endothelial inflammation and permeability.

MeSH terms

  • Aged
  • Aged, 80 and over
  • Atorvastatin / pharmacology*
  • Cells, Cultured
  • Coculture Techniques
  • Dexamethasone / pharmacology*
  • Dose-Response Relationship, Drug
  • Drug Therapy, Combination
  • Endothelial Cells / drug effects*
  • Endothelial Cells / metabolism
  • Endothelial Cells / pathology
  • Female
  • Hematoma, Subdural, Chronic / drug therapy*
  • Hematoma, Subdural, Chronic / metabolism
  • Hematoma, Subdural, Chronic / pathology
  • Humans
  • Kruppel-Like Transcription Factors / antagonists & inhibitors*
  • Kruppel-Like Transcription Factors / genetics
  • Kruppel-Like Transcription Factors / metabolism
  • Male
  • Middle Aged
  • Molecular Structure
  • Structure-Activity Relationship

Substances

  • KLF2 protein, human
  • Kruppel-Like Transcription Factors
  • Dexamethasone
  • Atorvastatin

Grants and funding

This work was supported by grants from the National Natural Science Foundation of China (grants 81671380, 81720108015, 81930031, and 81601996), Natural Science Foundation of Tianjin (grant 2017KJ193), Scientific Research Program Project of Tianjin Education Commission (2018ZD03), Tianjin Science and Technology Projects in Key Areas of Traditional Chinese Medicine (2018001), Beijing, Tianjin and Hebei special projects(19JCZDJC64600(Z)).