Erythritol is a commonly used non-nutritive sweetener with minimal effects on insulin and blood sugar levels. People with obesity, diabetes, and metabolic syndrome frequently use it as an alternative to sugar. Erythritol is produced in the human body through the pentose phosphate pathway and is also found naturally in fruits and fermented foods. Since its approval by the United States Food and Drug Administration (USFDA) in 2001, the consumption of erythritol has increased globally, primarily due to its inclusion in diet sodas and sugar-free foods.
The liver, kidneys, and erythrocytes all naturally create erythritol from glucose and fructose through the pathways of pentose phosphate. The study aimed to determine in vitro how the non-nutritive sweetener erythritol affected the production of endothelin (ET)-1, nitric oxide (NO), and oxidative stress in brain microvascular endothelial cells.
For three hours, human cerebral microvascular endothelial cells (hCMECs) were cultured and exposed to 6 mM erythritol, which is the same as the usual amount of erythritol (30 g) in an artificially sweetened beverage. The cells were maintained in endothelial growth medium supplemented with 100 μg/mL streptomycin and 100 U/mL penicillin. The supernatant was obtained after cells were cultivated for 24 hours with and without thrombin, both untreated and erythritol-treated, to measure the release of t-PA.
With the help of the Bio-Rad DC protein assay, whole-cell lysates were extracted from hCMECs to measure intracellular proteins, as per previous findings. The formation of reactive oxygen species (ROS) in response to 5 μM CellROX Deep Red Reagent was measured for 30 minutes at 37°C to assess intracellular oxidative stress. One-way analysis of variance (ANOVA) and repeated-measures ANOVA were used to identify differences in cellular outcome factors and t-PA release in response to thrombin.
In hCMECs, erythritol treatment enhanced intracellular ROS generation (204 ± 32% vs. 105 ± 4%), as well as levels of antioxidant proteins catalase (30.9 ± 0.3 vs. 24.4 ± 0.9 AU; P = 0.002) and superoxide dismutase-1 (332.1 ± 16.2 vs. 214.9 ± 4.7 AU; P = 0.002). Phosphorylated endothelial nitric oxide synthase (eNOS) levels were lower (52.1 ± 2.1 vs. 77.3 ± 9.1 AU; P < 0.001) while total eNOS expression was noted to be constant (102.8 ± 21.4 vs. 99.0 ± 19.9 AU). The synthesis of ET-1 has increased (34.6 ± 2.3 vs. 26.9 ± 1.5 pg/mL), and NO was decreased (5.8 ± 0.8 vs. 7.3 ± 0.7 µmol/L) in line with these molecular changes.
Thrombin-induced t-PA release was significantly reduced in erythritol-treated hCMECs (87.6 ± 8.3 vs. 110.2 ± 6.4 pg/mL in untreated cells), but intracellular t-PA levels and basal release were unchanged. The erythritol-induced oxidative load is not balanced by the increased levels of antioxidant enzymes, such as superoxide dismutase 1 (SOD-1) and catalase, which suggests a reactive cellular defense against oxidative stress. The study reveals impaired release of tissue-type plasminogen activator (t-PA), indicating a weakened blood-brain barrier (BBB) and diminished fibrinolytic capacity.
These findings align with epidemiological and clinical studies linking erythritol consumption to an increased risk of stroke and cardiovascular disease. The results highlighted the need for additional long-term clinical studies to evaluate the safety of erythritol and provide guidance for dietary recommendations, given its extensive use in sugar-free products.
Reference: Berry AR, Ruzzene ST, Ostrander EI, et al. The non-nutritive sweetener erythritol adversely affects brain microvascular endothelial cell function. J Appl Physiol. 2025;138:1571-1577. doi:10.1152/japplphysiol.00276.2025
