The Effect of Serum of Cerebral Small Vessel Disease Patients on Brain Endothelial Cells

Abstract

INTRODUCTIONCerebral Small Vessel Disease (CSVD) is characterized by perforating cerebral arterioles, venules and capillaries, and a consequent damage of cerebral white and deep grey matter. The pathogenesis of CSVD is unknown and could be asymptomatic in early phases. CSVD is a typical disease of elderly population and in stroke patients. However, the increasing use of MRI in last years, led to incidental findings in young people without classical vascular risk factors. To better understand the role of brain endothelium in onset of Cerebral Small Vessel Disease, we evaluated the effect of CSVD patients’ serum on human brain endothelial cells (HBEC‐5i).METHODSHBEC‐5i were treated for twenty‐four hours with 20% of serum from six CSVD patients or six age‐ and sex‐matched controls. Trypan blue exclusion method was used to evaluate proliferation rate and viability of treated cells compared to untreated cells. The effect of CSVD patients’ serum on HBEC‐5i, was evaluated by droplet digital PCR in terms of the mRNA expression of interleukins involved in inflammatory response(IL1β, IL1α, IL6, IL8, and IL10).RESULTSThe treatment with both patients’ and controls’ serum reduced the proliferation rate of cells, however no difference was observed between the two treatment. The viability, instead, was not influenced by serum treatment. The mRNA expression of pro‐inflammatory cytokines (IL1β, IL1α, IL6) was higher in HBEC‐5i treated with serum of CSVD patients compared to both untreated cells and cells treated with serum of controls. On the other hand, the mRNA expression of IL10 , an anti‐inflammatory molecule, increased only in cells treated with controls’ serum. None treatments affected the mRNA level of IL8.CONCLUSIONSIn brain endothelial cells, the serum of CSVD patients, differently from serum of controls, promotes the expression of pro‐inflammatory cytokines. These preliminary data suggest that inflammation could play a role in pathogenesis of CSVD.Support or Funding InformationThis project was supported by Grant Interreg ARTE (J22F170001005)