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Protective Effect of Tongluo Shenggu Capsule on Function Damage of Human Umbilical Vein Endothelial Cells Induced by Glucocorticoid / 中国实验方剂学杂志
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 48-55, 2021.
Article in Chinese | WPRIM | ID: wpr-906454
ABSTRACT

Objective:

To observe the effect of Tongluo Shenggu capsule (TLSGC) on glucocorticoid-induced vascular endothelial cell functional damage, and to preliminally explore the mechanism of action through MEK-ERK signaling pathway.

Method:

The blood vessel of aorta rings of normal SD rats were induced <italic>in vitro</italic> intervention with methylprednisolone sodium succinate (MPS, 0.04 g·L<sup>-1</sup>) and/or vascular endothelial growth factorVEGF, 20 μg·L<sup>-1</sup>), and were treated with TLSGC(12.5, 25, 50 μg·L<sup>-1</sup>) continuously for 5 days to observe the number, length and area of microvascular ring buds.In addition, human umbilical vein endothelial cells (HUVEC) induced by VEGF(20 μg·L<sup>-1</sup>) were added into MPS(0.04 g·L<sup>-1</sup>) and TLSGC (12.5, 25, 50 μg·L<sup>-1</sup>) were added. Then, Transwell migration, Transwell invasion and lumen formation experiments were used to detect the migration, invasion and lumen formation ability of HUVEC, respectively. The content of nitric oxide(NO) in the cell supernatant was detected by nitrate reductase method, the content of endothelin 1(ET-1) in the cell supernatant was detected by dry powder method. Moreover, the protein contents of vascular endothelial growth factor receptor 2 (VEGFR2), extracellular signal-regulated kinase (ERK), phospho-extracellular signal-regulated kinase (p-ERK), mitogen extracellular kinase1(MEK) and phosphorylated mitogen extracellular kinase1(p-MEK) in the cells were determined by Western blot.

Result:

Compared with the normal group, MPS could significantly inhibit the number, length and area of VEGF-induced rat thoracic aortic ring microvesselsHUVEC cell migration, invasion and lumen formation ability. It could reduce NO content and increase ET-1 content. MPS could also significantly reduce the protein content of VEGF-induced VEGFR2, p-MEK and p-ERK in HUVEC(<italic>P</italic><0.05,<italic>P</italic><0.01). Compared with the model group, TLSGC could dose-dependently increase the number, length and area of MPS-induced abnormally reduced rat thoracic aortic ring microvessels, promote MPS-induced abnormally decreased HUVEC cell migration, invasion and lumen formation ability. It could increase the protein contents of NO, VEGFR2, p-MEK and p-ERK in HUVEC, and reduce abnormally increased ET-1 content(<italic>P</italic><0.05<italic>,P</italic><0.01).

Conclusion:

TLSGC has a protective effect on the damage of angiogenesis and secretion of vascular endothelial cells induced by glucocorticoid, and the mechanism may be related to the activation of MEK/ERK signaling pathway.

Full text: Available Index: WPRIM (Western Pacific) Type of study: Prognostic study Language: Chinese Journal: Chinese Journal of Experimental Traditional Medical Formulae Year: 2021 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Type of study: Prognostic study Language: Chinese Journal: Chinese Journal of Experimental Traditional Medical Formulae Year: 2021 Type: Article