Objective: To investigate the effect of Tangeretin (TAN) on high glucose (HG)-induced apoptosis in
trophoblast cells by regulating the Ras homologous gene family member A (RhoA)/Rho-associated coiled-coil
forming protein kinase (ROCK) signaling pathway. Methods: Human chorionic trophoblast cells (HTR8-SVneo)
were cultured in vitro and randomly divided into control group (conventional culture), HG group (25 mmol/L
glucose), L-TAN, M-TAN, H-TAN groups (25, 50, and 100 μg/mL TAN, respectively), and PMA group (100 μg/mL
TAN+100 ng/mL RhoA/ROCK signaling pathway activator PMA). MTT method and colony formation assay were
applied to detect cell proliferation. ELISA was applied to detect the levels of interleukin (IL) 1β (IL-1β), tumor
necrosis factor-α (TNF-α), IL-6 and IL-8. Flow cytometry was applied to detect apoptosis. Western blotting was used
to detect the expression of proliferating cell nuclear antigen (PCNA), apoptosis-associated proteins (Bcl-2, Bax,
cleaved caspase-3), and proteins related to the RhoA/ROCK signaling pathway (RhoA, ROCK). Results: Compared
with the control group, the A490 of cells, clone number, the PCNA, Bcl-2 protein expression in the HG group
were decreased, while the IL-1β, TNF-α, IL-6, IL-8, apoptosis rate, cleaved caspase-3, Bax, RhoA, ROCK protein
expression were elevated. Compared with the HG group, the A490 of cells, clone number, the PCNA, Bcl-2 protein
expression in the L-TAN, M-TAN, and H-TAN groups
were increased, while the IL-1β, TNF-α, IL-6, IL-8,
apoptosis rate, cleaved caspase-3, Bax, RhoA, ROCK
protein expression were decreased. Compared withH-TAN group, the A490 of cells, clone number, the PCNA, Bcl-2 protein expression in PMA group were decreased,
while the IL-1β, TNF-α, IL-6, IL-8, apoptosis rate, cleaved caspase-3, Bax, RhoA, ROCK protein expression were
elevated. Conclusion: Tangeretin may attenuate high glucose-induced inflammatory responses in trophoblast cells by
inhibiting the RhoA/ROCK signaling pathway, thereby promoting cell proliferation and inhibiting cell apoptosis.
