Naoluo Xintong Decoction promotes proliferation of rat brain microvascular endothelial cells after oxygen-glucose deprivation by activating the HIF-1α/VEGF signaling pathway

doi:10.12122/j.issn.1673-4254.2025.09.17

Abstract

Abstract:

Objective To investigate the effects of Naoluo Xintong Decoction (NLXTD) on proliferation of rat brain microvascular endothelial cells (BMECs) after oxygen-glucose deprivation/reoxygenation (OGD/R) injury and role of the HIF-1α/VEGF pathway in mediating its effect. Methods Using a BMEC model of OGD/R, we tested the effects of 10% NLXTD-medicated rat serum, alone or in combination with 2ME2 or 10% NAKL, on cell proliferation, migration, tube-forming ability and permeability using CCK-8 assay, Transwell chamber assay, tube formation assay and permeability assay. Cellular expressions of VEGF and Notch were detected using ELISA and laser confocal immunofluorescence analysis, and the expressions of HIF-1α, VEGFR2, Notch1, ERK and P-ERK1/2 proteins were detected with Western blotting. Results OGD/R injury significantly decreased viability of BMECs. NLXTD treatment of the cells with OGD/R could significantly promoted cell proliferation, migration and tube formation ability, but these effects were strongly attenuated by application of 2ME2. NLXTD treatment also significantly increased the percentages of VEGF- and Notch-positive cells in the cell models and obviously enhanced the expression levels of HIF-1α, VEGFR2, Notch1 and P-ERK1/2. Conclusion NLXTD promotes proliferation, migration, and tube formation of rat BMECs after OGD/R injury possibly by activating the HIF-1α/VEGF signaling pathway.

Key words: ischemic stroke, Naoluo Xintong Decoction, HIF-1α/VEGF signaling pathway, oxygen-glucose deprivation/reoxygenation injury, cerebral microvascular endothelial cells

Yu ZHANG, Yinqi HU, Peipei LI, Xiao SHI, Wei XU, Jianpeng HU. Naoluo Xintong Decoction promotes proliferation of rat brain microvascular endothelial cells after oxygen-glucose deprivation by activating the HIF-1α/VEGF signaling pathway[J]. Journal of Southern Medical University, 2025, 45(9): 1980-1988.

Figures/Tables 6

Fig.1 Effect of NLXTD-medicated serum on viability of BMECs with OGD/R injury (n=6). ##P<0.01 vs normal group; &&P<0.01 vs OGD/R group.

Fig.2 Effect of NLXTD-medicated serum on migration capacity of BMECs with OGD/R injury (Original magnification: ×100, n=6). ##P<0.01 vs normal group; &&P<0.01 vs OGD/R group; *P<0.01 vs OGD/R+10%NLXTD group.

Fig.4 Effect of NLXTD-medicated serum on permeability of BMECs after OGD/R injury (n=6). ##P<0.01 vs normal group; &&P<0.01 vs OGD/R group;*P<0.05 vs OGD/R+10%NLXTD+2ME2 group.

Fig.5 Effect of NLXTD-medicated serum on supernatant VEGF level in BMECs after OGD/R injury. ##P<0.01 vs normal group; &&P<0.01 vs OGD/R group;*P<0.05 vs OGD/R+10%NLXTD group.

Fig.6 Effect of NLXTD-medicated serum on expressions of VEGF and Notch fluorescent protein in BMECs after OGD/R injury ( n=6). ##P<0.01 vs normal group; &&P<0.01 vs OGD/R group;*P<0.01 vs OGD/R+10%NLXTD group.

Fig.7 Effect of NLXTD-medicated serum on expressions of HIF-1a, VEGFR2, Notch1, ERK1/2, and P-ERK1/2 proteins in BMECs after OGD/R injury (n=4). ##P<0.05 vs normal group; &&P<0.01 vs OGD/R group; *P<0.01 vs OGD/R+10%NLXTD+2ME2 group.

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