Abstract
Background Vascular endothelial cell injury is not only the initiating factor of cardiovascular and cerebrovascular diseases but also the essence of blood stasis. Levistilide A (LA), a natural component isolated from the traditional Chinese herb, Ligusticum chuanxiong Hort, has traditional effects on improving blood circulation and removing stasis. In this study, the effects and potential mechanisms of LA in the rat model of blood stasis and the mechanism in endothelial cell injury have been explored. Materials and Methods In this experiment, the effects of LA on the model of acute blood stasis in rats were explored. The blood samples were collected for the measurement of coagulation and hemorheological indices, and the carotid arteries were also excised from rats for hematoxylin-eosin (HE) staining and immunohistochemistry (IHC). In addition, the improvement effects of LA on the H2O2-induced human umbilical vein endothelial cell (HUVEC) injury model were evaluated. And the cell viability detection was conducted by the CCK8 assay, and the pathway-related protein expressions were detected by western blotting. Results In vivo, compared with the model group, the treatment of LA (10 mg/kg) could reduce the FIB (fibrinogen) content (P < 0.01), increase the INR (international normalized ratio) and PT (prothrombin time) (P < 0.01), and reduce the plasma viscosity (P < 0.05) and whole blood viscosities of low, medium, and high shear rates in the blood of blood stasis model rats (P < 0.01). In vitro, the cell viability in the LA-pretreated group was higher than that of the model group (P < 0.05). The expression levels of PI3K, AKT, and eNOs in the LA-pretreated group were increased (P < 0.01) as compared to the model group. Conclusion These findings demonstrated that LA has the ability to improve blood hypercoagulation and blood viscosity, and enhance the viability of cells. It is more likely that it exerts a protective effect on the endothelial cell through the PI3K-AKT-eNOs pathway. These results indicate LA will be a potential candidate to cure blood stasis with endothelial cell injury.
Highlights
Coagulation balance is maintained by antithrombin (AT), thrombomodulin (TM), tissue factor pathway inhibitor (TFPI), nitric oxide (NO), endothelin (ET), etc., emphasizing the nature complexities of activities governed by endothelial cells that serve as the main site for their synthesis and release [5, 6]
An early sign of changing vascular homeostasis can be used as a predictor of cardiovascular events related to the development of cardiovascular and cerebrovascular diseases [7, 8]. e Consensus on the Diagnosis and Treatment of Blood Stasis with Integrated Traditional Chinese and Western Medicine proposes that blood rheology and coagulation function, NO and ET levels, platelet aggregation, adhesion, and abnormalities captured in the images can be used as diagnostic indicators for blood stasis, and clearly advocates that endothelial cell function damage is the essence of blood stasis [9]. is experiment mainly focuses on the interdependence and mutual influence between eNOs and endothelial cell functions
We have verified the relationship between blood stasis and hemorheology and endothelial cell injury through in vivo and in vitro experiments, and verified that Levistilide A (LA) can improve the abnormality of the indicators above to a certain extent and primarily explored the mechanism of protection via the PI3K-AKT signaling pathway
Summary
Vascular endothelial cell injury is the initiating factor of cardiovascular and cerebrovascular diseases and the essence of blood stasis. The effects and potential mechanisms of LA in the rat model of blood stasis and the mechanism in endothelial cell injury have been explored. In this experiment, the effects of LA on the model of acute blood stasis in rats were explored. The improvement effects of LA on the H2O2-induced human umbilical vein endothelial cell (HUVEC) injury model were evaluated. E expression levels of PI3K, AKT, and eNOs in the LA-pretreated group were increased (P < 0.01) as compared to the model group. It is more likely that it exerts a protective effect on the endothelial cell through the PI3K-AKT-eNOs pathway. Ese results indicate LA will be a potential candidate to cure blood stasis with endothelial cell injury It is more likely that it exerts a protective effect on the endothelial cell through the PI3K-AKT-eNOs pathway. ese results indicate LA will be a potential candidate to cure blood stasis with endothelial cell injury
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