Goreisan inhibits vascular endothelial cell migration and angiogenesis

Abstract

Aim Goreisan (GRS) is a Japanese Kampo medicine, which has been clinically used to treat edema and headache. Recently, a clinical study indicated that GRS significantly suppresses the recurrence of chronic subdural hematoma (CSDH). In CSDH, leakage from immature blood vessels on the neomembrane is involved in the recurrence or exacerbation. Although several clinical reports have shown the effectiveness of GRS, the pharmacological properties and underlining mechanism of GRS are not clear. In this study, we examined the effect of GRS on the migration and proliferation of vascular endothelial cells via in vitro and in vivo experiments. Method Human umbilical vascular endothelial cells (HUVECs) were treated with vascular endothelial growth factor (VEGF) for 24 h, and migration ability, extracellular signal‐regulated kinase (ERK) phosphorylation level, and aquaporin‐1 (AQP1) expression level were measured. Matrigel containing VEGF was injected subcutaneously into the abdomen of C57BL mice (Matrigel plug assay). After oral administration of GRS for seven days, the effect on angiogenesis was examined by immunostaining. Results GRS inhibited VEGF‐induced migration in a dose‐dependent manner. GRS also inhibited VEGF‐induced phosphorylation of ERK, which is a major signaling molecule for the migration. In addition, GRS markedly decreased protein and mRNA expression of AQP1, which is another important regulator of endothelial cell migration. Finally, GRS inhibited angiogenesis, as revealed by in vivo Matrigel plug assay. Conclusion The findings indicated that GRS can suppress angiogenesis through inhibiting the migration of endothelial cells by downregulating ERK activation and aquaporin‐1 expression, leading to the preventive effect of GRS against CSDH recurrence.

Impact of Goreisan components on rat mesenteric collecting lymphatic vessel pumping

“…It is also worth noting that in the current study, we observed that Goreisan administration to rats for 1 week did not cause a significant change in AQP‐1 protein levels in mesenteric lymphatic vessels. A previous report shows that treatment of human umbilical vein endothelial cells (HUVEC) with 0.1 mg/mL Goreisan for 24 h attenuates VEGF‐induced increases in AQP‐1 mRNA and protein expression 32 . In addition, administration of Goreisan to mice for 1 week decreased angiogenesis in a Matrigel plug assay 32 .…”

“…A previous report shows that treatment of human umbilical vein endothelial cells (HUVEC) with 0.1 mg/mL Goreisan for 24 h attenuates VEGF‐induced increases in AQP‐1 mRNA and protein expression 32 . In addition, administration of Goreisan to mice for 1 week decreased angiogenesis in a Matrigel plug assay 32 . While our results appear to be in contrast to these previous findings, it is difficult to directly compare our in vivo findings to the in vitro findings.…”

“…In addition, the route of administration of Goreisan differed between the current study (oral gavage) and the Matrigel plug assay previously reported, in which the Goreisan appears to have been directly administerd in the implantation. 32 It is also possible that with a larger sample size, we might find that oral Goreisan administration decreases AQP-1 levels in mesenteric lymphatics, and this will be taken into consideration in future studies.…”

“…32 In addition, administration of Goreisan to mice for 1 week decreased angiogenesis in a Matrigel plug assay. 32 While our results appear to be in contrast these previous findings, it is difficult to directly compare our in vivo findings to the in vitro findings. In addition, the route of administration of Goreisan differed between the current study (oral gavage) and the Matrigel plug assay previously reported, in which the Goreisan appears to have been directly administerd in the implantation.…”

Uncovering the metabolomic effects of Kampo formulas using 1H‐NMR spectroscopy: A case study of goreisan

“…GRS has both antidiarrhea and tissue-protective effects through inhibiting inflammation-induced AQP3 expression in the intestine [3]. GRS also suppresses angiogenesis by inhibiting migration of endothelial cells and downregulating extracellular signal-regulated kinase (ERK) activation and AQP1 expression, with a preventive effect against chronic subdural hematoma (CSDH) recurrence [4], and reduces the recurrence rate of CSDH [5]. Additionally, GRS has been found to inhibit AQP4 activity and thus has the potential to reduce brain edema after CSDH and other brain injuries [6,7].…”