Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2020 Aug 12:14:3291-3299.
doi: 10.2147/DDDT.S256050. eCollection 2020.

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

Yueshan Fan #  1   2   3 , Dong Wang #  1   2 , Chenxu Rao #  1   2   3 , Ying Li  1   2 , Hongtao Rong  1   2 , Zengguang Wang  1   2 , Jianning Zhang  1   2
Affiliations

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

Yueshan Fan et al. Drug Des Devel Ther. .

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.

PubMed Disclaimer

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest and report no conflicts of interest for this work.

Figures

Figure 1
Figure 1
The expression of KLF-2 in endothelial cells was decreased after hematoma sample injury. (A) The morphological changes in cells at different time points after stimulation with hematoma supernatant observed by HPICM; (B) The changes in KLF-2 expression after treatment of hematoma supernatant-damaged cells with different interventions; (C) Gray value analysis of panel (B); (D) The changes in the KLF-2 mRNA level after treatment of hematoma-damaged cells with different interventions. *P < 0.05; **P < 0.01; ***P < 0.001.
Figure 2
Figure 2
Hematoma sample stimulates cocultured cells and induces changes in the expression of tight junction proteins after knockdown of KLF-2 in endothelial cells. (A) The changes in tight junction proteins in endothelial cells after injury observed by immunofluorescence staining; (B) Western blot analysis of the changes in tight junction protein expression in endothelial cells with different genotypes after injury; (C). Gray value analysis of panel B. *P < 0.05.
Figure 3
Figure 3
Hematoma sample stimulates cocultured cells and induces changes in the expression of vascular inflammation markers after ablation of KLF-2 in endothelial cells. (A) Western blot analysis of the changes in adhesion protein expression in endothelial cells with different genotypes after injury; (B) Gray value analysis of panel (A); (C) PCR analysis of the changes in adhesion protein mRNA levels in endothelial cells with different genotypes after injury; (D) ELISA analysis of the changes in inflammatory factor and VEGF expression in endothelial cells with different genotypes after injury. *P < 0.05, **P<0.01.
Figure 4
Figure 4
Hematoma sample stimulates cocultured cells and induces changes in the expression of vascular inflammation markers and tight junction proteins after ablation of KLF-2 in endothelial cells given different treatments. (A) Western blot analysis of the changes in tight junction and adhesion protein expression in endothelial cells with different genotypes given different treatments after injury; (B) Gray value analysis of panel (A); (C) PCR analysis of the changes in the mRNA levels of tight junction and adhesion proteins in endothelial cells with different genotypes given different treatments after injury; (D) ELISA analysis of the changes in inflammatory factor and VEGF expression in endothelial cells with different genotypes given different treatments after injury. *P < 0.05.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Chen JC, Levy ML. Causes, epidemiology, and risk factors of chronic subdural hematoma. Neurosurg Clin N Am. 2000;11(3):399–406. doi:10.1016/S1042-3680(18)30101-3 - DOI - PubMed
    1. Allison A, Edlmann E, Kolias AG, et al. Statistical analysis plan for the Dex-CSDH trial: a randomised, double-blind, placebo-controlled trial of a 2-week course of dexamethasone for adult patients with a symptomatic chronic subdural haematoma. Trials. 2019;20(1):698. doi:10.1186/s13063-019-3866-6 - DOI - PMC - PubMed
    1. Almenawer SA, Farrokhyar F, Hong C, et al. Chronic subdural hematoma management: a systematic review and meta-analysis of 34,829 patients. Ann Surg. 2014;259(3):449–457. doi:10.1097/SLA.0000000000000255 - DOI - PubMed
    1. Miah IP, Holl DC, Peul WC, et al. Dexamethasone therapy versus surgery for chronic subdural haematoma (DECSA trial): study protocol for a randomised controlled trial. Trials. 2018;19(1):575. doi:10.1186/s13063-018-2945-4 - DOI - PMC - PubMed
    1. Holl DC, Volovici V, Dirven CMF, et al. Pathophysiology and Nonsurgical Treatment of Chronic Subdural Hematoma: from Past to Present to Future. World Neurosurg. 2018;116(402–411):e402. doi:10.1016/j.wneu.2018.05.037 - DOI - PubMed

MeSH terms