Role of Endothelial Nrf2 in the Regulation of Vasomotor Capacity and Arterial Blood Pressure
Authors: Dhyani Neha , Rudebush Tara L , Anderson Cody , Allen Michael , Gao Lie , Zucker Irving H , Park Song Young 
Published: May 2024

Introduction: Hypertension is a serious global public health issue. It is a complex disease with multiple factors contributing to its pathophysiology. One of these factors is oxidative stress, which results from an imbalance between the production and elimination of reactive oxygen species (ROS). Studies have shown that an increased ROS in endothelial cells (EC) impairs the vasodilatory capacity of blood vessels that contributes to blood pressure elevation. However, the modulation of endothelial ROS in the setting of hypertension is still not completely understood. To maintain cellular homeostasis, antioxidant enzymes continually adjust ROS levels under the orchestrating of transcription factor Nuclear factor erythroid 2-related factor 2 (Nrf2), a master regulator of endogenous antioxidant responses. Modulation of endothelial Nrf2 expression may be a potential therapeutic option in hypertension. Therefore, the major objective of this study was to evaluate the role of Nrf2 in the modulation of endothelium associated vascular function and how it regulates vascular tone and blood pressure (BP). Methods: BP was measured in the conscious state using radiotelemetry transducers implanted into EC-specific Nrf2-knockout mice (Tie2-Cre-Nrf2 Floxed), EC specific Keap1 knockout mice (Tie2-Cre-Keap1 Floxed) and Wildtype (WT), all on the C57B6J background. After baseline BP recordings, mice were implanted with subcutaneous osmotic minipumps (7 days) for Angiotensin II (Ang II) (600ng/Kg/min) infusion or in a separate group L-nitroarginine methyl ester (L-NAME) (100 mg/kg/d in drinking water) for 2 weeks. For each intervention, recordings were done daily for 2 hours and one 24-hour measurement was done once per week. Endothelial specific downregulation or upregulation of Nrf2 was confirmed by immunohistochemistry on mouse aortic tissue and protein expression from mouse EC. For in-vitro studies: femoral-to-popliteal artery segments were harvested from mice of each genotype. Vasodilation was assessed in response to flow (30 uL·min−1) and acetylcholine (ACh, 10−7-10−3 M) with and without L-NAME, using video microscopy. Results: Ang II infusion significantly increased the mean arterial pressure (MAP) in each group when compared to their baseline values (Baseline: Nrf2KO = 92.6 ± 3.1, Keap1KO = 93.8 ± 4.1 and WT = 92.8 ± 1.7 mmHg; after Ang II Nrf2KO = 141.7 ± 3.8, Keap1KO = 121.9±5.4 and WT = 120.4±5.4 mmHg). The response to ANG II was significantly (p<0.05) augmented in Nrf2KO mice and stayed elevated for a longer period after emptying of the pump. The response to L-NAME was similar in both Nrf2KO and WT mice. Protein expression of Nrf2 was decreased in Nrf2KO mice in comparison to WT mice. Nrf2KO mice exhibited reduced endothelium-dependent vasodilation induced by flow (WT: 35.8± 1.3%, Nrf2KO: 24.1 ±1.7%; p<0.0001) and ACh (10−3M) (WT: 60.4±4.1%, Nrf2KO: 44.7± 2.1%; p<0.0001) compared to WT. In contrast, endothelial dependent vasodilatory responses to flow and Ach were markedly elevated in Keap1KO mice (51.3± 6.8%, p<0.0001; 71.6±11.6%, p<0.0087, respectively) compared to both WT and Nrf2KO mice. Conclusion: These results suggest that antioxidant defense mediated by Nrf2 in the endothelium plays a significant role in the modulation of blood pressure and vascular tone. Based on the L-NAME data, these effects are most likely due to an increase in the nitric oxide bioavailability and reduced ROS. University of Nebraska Theodore Hubbard foundation. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.