大黄蒽醌类提取物对脓毒症小鼠中性粒细胞胞外诱捕网形成的影响OA

Objective To investigate the effect of Rhubarb anthraquinone extracts on neutrophil extracellular traps(NETs)formation in septic mice and to explore the underlying mechanisms.Methods Animal experiment:Forty male Balb/c mice were randomly divided into a control group(CG),a lipopolysaccharide(LPS)group,a LPS+free anthraquinone group(FA),and a LPS+combined anthraquinone group(CA)using a random number table,with 10 mice in each group.Twelve hours before LPS treatment,the LPS+FA group and the LPS+CA group were administered with FA and CA(100 mg/kg each)via gavage,respectively,while the CG group and the LPS group were administered with 0.5%sodium carboxymethyl cellulose solution via gavage.Twelve hours later,the LPS group,the LPS+FA group,and the LPS+CA group received LPS(15 mg/kg)via an intraperitoneal injection,inducing a sepsis model.Simultaneously,the CG group received 0.9%sterile saline of an equivalent volume.Post-modeling,the LPS+FA group and the LPS+CA group received treatment with their corresponding extracts(100 mg/kg each)via gavage,while the CG group and the LPS group were re-administered 0.5%sodium carboxymethyl cellulose solution of an equal volume.Then,serum tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)levels were measured using enzyme-linked immunosorbent assay(ELISA).Histopathological changes in liver and ileum tissues were observed via hematoxylin and eosin(HE)staining.Citrullinated histone H3(CitH3)and myeloperoxidase(MPO)expression levels in the liver and ileum were detected by Western blot(WB).Cell experiment:HL-60 cells were treated with dimethyl sulfoxide for 5 days,induced into mature granulocytes.Following maturation verification,these cells were divided into a CG group,a phorbol 12-myristate 13-acetate(PMA)group,a diphenyleneiodonium chloride(DPI)+PMA group,a FA+PMA group,and a CA+PMA group.Before PMA intervention,the FA+PMA group,the CA+PMA group,and the DPI+PMA group were pretreated with FA(800 μmol/L),CA(750 μmol/L),and DPI(10 μM),respectively,for 30 min.Subsequently,these groups were intervened with PMA for 4 h,after which the medium was replaced and incubation continued for another 20 h.Then,cellular reactive oxygen species(ROS)levels were assessed using flow cytometry,while CitH3 expression level was evaluated by immunofluorescence staining.Results Animal experiment:Compared with the CG group,the LPS group exhibited significantly elevated serum TNF-α and IL-1β expression levels(P<0.0001),aggravated pathological damage in the liver and ileum,alongside increased CitH3 and MPO expression levels(P<0.01,P<0.001,P<0.0001).Compared with the LPS group,the LPS+FA group and the LPS+CA group showed decreased serum TNF-α and IL-1β expression levels(P<0.001,P<0.0001),ameliorated histopathological damage in the liver and ileum,along with reduced CitH3 and MPO expression levels(P<0.05,P<0.01,P<0.001).Cell experiment:Compared with the CG group,the PMA group demonstrated significantly increased ROS and CitH3 release(P<0.01,P<0.001).Compared with the PMA group,ROS release and the expression of CitH3 were decreased in the FA+PMA and the CA+PMA group(P<0.01,P<0.001).Conclusion FA and CA can inhibit the NETs formation in septic mice,with a potential mechanism related to reduced ROS release.