Research Article
Shenfu Injection Alleviating OGD-Induced Brain Microvascular Endothelial Cell Injury via Inhibiting eNOS Uncoupling
Issue:
Volume 14, Issue 6, December 2026
Pages:
129-139
Received:
1 October 2026
Accepted:
27 October 2026
Published:
10 November 2026
DOI:
10.11648/j.ajcem.20261406.11
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Abstract: Objective: The pathological process of vascular dementia originates from functional impairment of brain microvascular endothelial cells under ischemia‑hypoxia conditions, and endothelial nitric oxide synthase (eNOS) uncoupling serves as a critical hallmark of endothelial dysfunction. This study aimed to explore whether the protective effect of Shenfu injection against oxygen‑glucose deprivation (OGD)‑induced injury in brain microvascular endothelial cells involves eNOS uncoupling. Methods: An OGD model of bEnd.3 cells was induced by sodium dithionite treatment. Experimental groups included control group, OGD group, low‑dose Shenfu group (0.1mg/mL), medium‑dose Shenfu group (0.5mg/mL), and high‑dose Shenfu group (1.0mg/mL). MTT assay and lactate dehydrogenase (LDH) detection were performed to evaluate cell viability and injury. Griess assay was adopted to measure nitric oxide (NO) concentration in cell supernatants. Enzyme‑linked immunosorbent assay (ELISA) was used to detect intracellular cyclic guanosine monophosphate (cGMP) levels. Western blot was applied to determine the protein expression of eNOS. Reactive oxygen species (ROS) content was quantified with DCFH‑DA fluorescent probe. Xanthine oxidase assay was utilized for superoxide dismutase (SOD) activity measurement, and DTNB method was used to detect reduced glutathione (GSH) content. Results: Compared with the control group, the OGD group exhibited marked cellular morphological damage, significantly reduced cell viability, elevated LDH release and ROS levels (P<0.01); the contents of NO and cGMP decreased significantly (P<0.01); eNOS protein expression showed no statistically significant difference (P>0.05); meanwhile, antioxidant indicators including SOD activity and GSH content declined remarkably (P<0.01). Compared with the OGD group, Shenfu injection produced dose‑dependent improvements in above‑mentioned parameters: cell viability was significantly increased while LDH activity was lowered (P<0.01); NO and cGMP contents were elevated (P<0.01), without significant alteration in eNOS protein expression (P>0.05); additionally, intracellular ROS level was reduced (P<0.01), accompanied by increases in SOD activity and GSH content (P<0.01). Conclusion: Shenfu injection mitigates OGD‑induced injury of brain microvascular endothelial cells. Its underlying mechanism may be associated with potentiating the antioxidant capacity of SOD and GSH, eliminating excessive ROS, inhibiting eNOS uncoupling, and further improving NO bioavailability.
Abstract: Objective: The pathological process of vascular dementia originates from functional impairment of brain microvascular endothelial cells under ischemia‑hypoxia conditions, and endothelial nitric oxide synthase (eNOS) uncoupling serves as a critical hallmark of endothelial dysfunction. This study aimed to explore whether the protective effect of Shen...
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,
Jiazilin Hu
