Abstract
In this study, we investigated the protective effects of gastrodin (Gas) against homocysteine‐induced human umbilical vein endothelial cell (HUVEC) injury and the role of the phosphoinositide 3‐kinase (PI3K)/threonine kinase 1 (Akt)/endothelial nitric oxide synthase (eNOS) and NF‐E2‐related factor 2 (Nrf2)/antioxidant response element (ARE) pathways. We stimulated cells with homocysteine (1 mmol/L, 24 hours) and tested the effects of gastrodin (200‐800 μg/mL) on cell viability and the production of malondialdehyde (MDA), lactate dehydrogenase (LDH) and reactive oxygen species (ROS). Then, Nrf2 distribution in the cytoplasm and nucleus as well as the expression of enzymes downstream of Nrf2 was determined. Furthermore, we analysed the expression of bax, bcl‐2 and cleaved caspase3, and assessed the involvement of the PI3K/Akt/eNOS pathway by Western blots. Finally, we tested the vasoactive effect of gastrodin in thoracic aortic rings. The results showed that gastrodin decreased MDA, LDH and ROS production and increased cell viability, NO production and relaxation of thoracic aortic rings. Moreover, the protective effects of Gas on NO production and relaxation of thoracic aortic rings were blocked by L‐NAME but enhanced by Cav‐1 knockdown, and MK‐2206 treatment abolished the effect of Gas on the ROS. In addition, treatment with gastrodin increased Nrf2 nuclear translocation, thus enhancing the expression of downstream enzymes. Finally, gastrodin increased the expression of PI3K, p‐Akt, and eNOS and decreased Cav‐1 protein expression. In conclusion, our study suggested that gastrodin may protect HUVECs from homocysteine‐induced injury, and the PI3K/Akt/eNOS and Nrf2/ARE pathways may be responsible for the efficacy of gastrodin.
Highlights
The results showed that Hcy decreased the intracellular levels of ATP in human umbilical vein endothelial cell (HUVEC), which were restored by Gas in a concentration-dependent manner, and the 400 and 800 μg/mL Gas groups showed significant differences compared with the model group (Figure 1B)
The results showed that the stimulation of HUVECs with Hcy decreased the expression of HO-1, SOD-1 and catalase; Gas dose-dependently increased the expression of HO-1, SOD-1 and catalase at the protein and mRNA levels, and all these parameters showed significant differences in the 400 and 800 μg/mL Hcy groups but not in the 200 μg/ mL Hcy group, indicating that low-dose Gas had little effect on the expression of NF-E2-related factor 2 (Nrf2)/antioxidant response element (ARE) downstream enzymes (Figure 2C, D)
(5) Inhibition of the phosphoinositide 3-kinase (PI3K)/Akt pathway abolished the protective effects of Gas. (6) Hcy addition significantly decreased the relaxation of thoracic aortic rings, which was improved by Gas through the regulation of Cav-1/endothelial nitric oxide synthase (eNOS)
Summary
Experimental and clinical studies have proven that homocysteine (Hcy) plays an important role in the development of various cardiovascular diseases.[1]. The Hcy-induced reduction of the Nrf2-dependent antioxidant defence system suppressed antioxidant enzymes downstream of Nrf2.9,10 research has indicated that decreasing the accumulation of ROS and restoring NO production could inhibit Hcy-induced injury.[11]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
