Aim. Ginkgolide B is a Ginkgo biloba leaf extract that has been identified as a natural platelet‐activating factor receptor (PAFR) antagonist. We investigated the effect of ginkgolide B on high glucose‐induced TLR4 activation in human umbilical vein endothelial cells (HUVECs). Methods. Protein expression was analyzed by immunoblotting. Small‐interfering RNA (siRNA) was used to knock down PAFR and TLR4 expression. Results. Ginkgolide B suppressed the expression of TLR4 and MyD88 that was induced by high glucose. Ginkgolide B also reduced the levels of platelet endothelial cell adhesion molecule‐1, interleukin‐6, and monocyte chemotactic protein 1. Further, we examined the association between PAFR and TLR4 by coimmunoprecipitation. The result showed that high glucose treatment caused the binding of PAFR and TLR4, whereas ginkgolide B abolished this binding. The functional analysis indicated that PAFR siRNA treatment reduced TLR4 expression, and TLR4 siRNA treatment decreased PAFR expression in high glucose‐treated HUVECs, further supporting the coimmunoprecipitation data. Ginkgolide B inhibited the phosphorylation of Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) and p38 mitogen‐activated protein kinase (MAPK). Conclusion. Ginkgolide B exerted protective effects by inhibiting the TLR4‐mediated inflammatory response in high glucose‐treated endothelial cells. The mechanism of action of ginkgolide B might be associated with inhibition of the JAK2/STAT3 and p38 MAPK phosphorylation.

Introduction The complex pathophysiology and diverse clinical manifestations of sepsis and septic shock continue to make early diagnosis and severity assessment challenging. Previous studies revealed the distinct roles of histone H3 lysine 18 lactylation (H3K18la) and H3 lysine 18 acetylation (H3K18ac) in infection. However, the functions and interactions of these modifications remain unclear. This study aimed to investigate the expression and roles of H3K18la and H3K18ac in patients with sepsis and septic shock. Methods This ambispective cohort study enrolled 86 critically ill patients (13 sepsis, 37 septic shock, and 36 noninfectious) and 12 healthy volunteers. Baseline information and laboratory data were collected. H3K18la and H3K18ac levels in peripheral blood mononuclear cells were detected via Western blotting. Serum cytokines, arginase-1 (ARG1), and Krüppel-like factor 4 (KLF4) mRNA were assayed via microsphere immunofluorescence and quantitative real-time polymerase chain reaction. The potential value of H3K18la, H3K18ac, and their ratio (H3K18la/ac) in the diagnosis and severity assessment was analyzed using logistic regression, receiver operating characteristic curve, and correlation analysis. Results Compared with the noninfectious group, the infectious group presented increased H3K18la and H3K18la/ac and decreased H3K18ac levels, with H3K18la/ac as an independent diagnosis biomarker. Compared with the sepsis group, the septic shock group presented higher H3K18la and H3K18la/ac and lower H3K18ac levels. H3K18la and H3K18la/ac levels correlated positively with sequential organ failure assessment (SOFA) scores, length of intensive care unit (ICU) stay, and mechanical ventilation time. H3K18ac levels correlated negatively with SOFA scores and mechanical ventilation time. H3K18la levels correlated negatively with interferon-α (IFN-α) and interleukin‑5 (IL‑5) and positively with IL-10 expression. H3K18ac levels correlated negatively with IL-6, IL-1β, IL-8, and IL-10 expression. H3K18la/ac levels correlated negatively with IFN-α and IL-5 and positively with IL-6, IL-8, and IL-10 expression. H3K18la and H3K18la/ac correlated positively, whereas H3K18ac correlated negatively with ARG1 and KLF4 mRNA expression. Conclusions H3K18la, H3K18ac, and H3K18la/ac can serve as biomarkers for the diagnosis and severity assessment of sepsis and septic shock through their involvement in inflammatory responses and macrophage polarization, thereby informing targeted therapies to modulate immune responses and improve patient outcomes.

Endothelial cells are sensitive to changes in both blood components and mechanical stimuli. Endothelial cells may undergo phenotypic changes, such as changes in adhesion protein expression, under different shear stress conditions. Such changes may impact platelet and monocyte adhesion to endothelial cells. This phenomenon is linked to chronic vascular inflammation and the development of atherosclerosis. In the present study, we investigated the effects of ginkgolide B on platelet and monocyte adhesion to human umbilical vein endothelial cells (HUVECs) under different conditions of laminar shear stress. Platelet and monocyte adhesion to endothelial cells was determined by the Bioflux 1000. HUVECs were incubated with ginkgolide B or aspirin for 12 h, and then TNFα was added for 2 h to induce the inflammatory response under conditions of 1 and 9 dyn/cm2 laminar shear stress. The protein expression was analyzed by Western blot. The number of platelets that adhered was greater under conditions of 1 dyn/cm2 than under conditions of 9 dyn/cm2 of laminar shear stress (74.8 ± 19.2 and 59.5 ± 15.1, respectively). Ginkgolide B reduced the tumor necrosis factor α (TNFα)-induced increase in platelet and monocyte adhesion to HUVECs at 1 and 9 dyn/cm2 of laminar shear stress. In TNFα-treated HUVECs, the number of monocytes that adhered was greater under conditions of 1 dyn/cm2 of laminar shear stress compared with 9 dyn/cm2 (29.1 ± 4.9 and 22.7 ± 3.7, respectively). Ginkgolide B inhibited the TNFα-induced expression of vascular cell adhesion molecule-1(VCAM-1), VE-cadherin, and Cx43 in HUVECs at 1 and 9 dyn/cm2. The expression of these proteins was not different between 1 and 9 dyn/cm2. Ginkgolide B suppressed platelet and monocyte adhesion under different conditions of laminar shear stress. Moreover, ginkgolide B reduced VCAM-1, VE-cadherin and Cx43 expression in TNFα-treated HUVECs under laminar shear stress. This suggested that ginkgolide B might shed light on the treatment of inflammation in atherosclerosis.