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KMID : 0363320200410040553
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Journal of Korean Oriental Internal Medicine
2020 Volume.41 No. 4 p.553 ~ p.562
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Mitigative Effect of Sipjeondaebo-tang on RhoA Activation in Cold-Exposed Vascular Cells
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Lee Kang-Wook
Kim Myeong-Sun
Kim Yun-Gyung
Hwang Hyun-Ha
Go Ho-Yeon
Sun Seung-Ho
Choi You-Kyung
Yang Seung-Bo
Song Yun-Kyung
Jeon Chan-Yong
Ko Seong-Gyu
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Abstract
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Objective: To investigate the vasodilatory effect of Sipjeondaebo-tang by inhibiting RhoA activity in vascular cells during cold exposure.
Methods: Human vascular endothelial cells and pericytes were pretreated with Sipjeondaebo-tang for 30 min, followed by incubation at 37 ℃ (control) or 25 ℃ (cold exposure) for 30 min. Activation of endothelin-1-mediated RhoA in pericytes was assessed by pretreating the cells with Sipjeondaebo-tang for 30 min, followed by incubation with endothelin-1 at 37 ℃ for 30 min. Western blotting was performed to measure the expression of active RhoA. Endothelin-1 and nitric oxide release from endothelial cells was examined with enzyme-linked immunosorbent assay kits. The formation of stress fibers and focal adhesion complexes was analyzed by immunocytochemistry.
Results: Cold treatment activated RhoA in both pericytes and vascular endothelial cells, whereas Sipjeondaebo-tang treatment inhibited this activation. Sipjeondaebo-tang treatment also reversed the cold-mediated production of endothelin-1 and nitric oxide. Cold exposure promoted the formation of stress fibers and focal adhesion complexes by increasing the expression of phospho-focal adhesion complex kinase, whereas Sipjeondaebo-tang treatment suppressed this response.
Conclusions: These findings suggested that Sipjeondaebo-tang inhibits cold-induced RhoA activation and its related pathway components, including endothelin-1 and nitric oxide, in vascular cells. Therefore, Sipjeondaebo-tang could be beneficial for the treatment of Raynaud’s phenomenon.
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KEYWORD
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Raynaud’s phenomenon, cold, rhoA, endothelin-1, nitric oxide, Sipjeondaebo-tang
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