Fufang Xueshuantong protects retinal vascular endothelial cells from high glucose by inhibiting YAP

[Speaker] Jing Han:1
[Co-author] Zhenglin Wang:1, Wei Xing:1, Yongli Song:1, Yan Wu:1
1:Beijing University of Chinese Medicine, China

Background: Fufang Xueshuantong (FXST), which is composed of Panax notoginseng, Salvia miltiorrhiza, Astragalus membranaceus and Scrophularia ningpoensis, has been reported to treat diabetic retinopathy (DR), but the underlying mechanism of FXST is not determined. Previous results showed that FXST could modulate the protein levels of YAP in retinas of diabetic rats. Thus the purpose of this study is to investigate if the pharmacological effect of FXST is dependent on YAP.
Methods: Retinal vascular endothelial cells (RVEC) were assigned into the normal group (5.5 mM glucose), high glucose group (25 mM glucose) and FXST group (25 mM glucose + different doses of FXST). After the cells were treated with different medium for three days, the CCK-8 assay, transwell assay, and tube formation assay were used to observe the effect of FXST on endothelial cells. Western blot method was used to examine the protein expression of the YAP in the RVEC. Then sh-YAP bearing plasmid was prepared and transfected into the RVEC. The pharmacological effect of FXST was evaluated again in different circumstances (FXST+ high glucose, sh-YAP+FXST+high glucose, sh-NC+FXST+high glucose).
Results: Compared with the normal group, the cell viability, the number of migrated cells, and the tube length were increased in high glucose group (P<0.01); compared with the high glucose group, the cell viability, the number of migrated cells, and the total length were inhibited by FXST (P<0.05 or P<0.01). The Western blot results showed that, in comparison to the normal group, the high glucose group showed higher expression of YAP (P<0.05); In comparison to the high glucose group, FXST inhibited the expression of YAP (P<0.05). After the sh-YAP was transfected into the RVEC, the content of YAP was remarkably decreased compared with sh-NC group. In addition, the number of migrated cells and the total length were elevated in sh-YAP+FXST+ high glucose group compared with FXST+ high glucose group, but there were not significant differences between FXST+ high glucose group and sh-NC+FXST+ high glucose group.
Conclusion: FXST could protect RVEC from high glucose, and this effect was mediated by inhibiting YAP protein.
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