摘要
目的 研究血必净注射液调控高迁移率族蛋白1(HMGB1)/Toll样受体4(TLR4)/核因子-κB(NF-κB)通路对脓毒症小鼠肺损伤的保护作用.方法 雄性C57BL/6小鼠随机分为对照组,模型组和低、中、高剂量血必净组,阴性对照(NC)组,NC+模型组,NC+高剂量血必净组,HMGB1+高剂量血必净组.采用盲肠结扎穿孔术建立脓毒症肺损伤模型,造模前给予NC慢病毒或HMGB1慢病毒尾静脉注射,造模当天给予血必净注射液(剂量5、10、15 mL/kg)腹腔注射,2次/d,连续3 d,末次给药后24 h进行取材和检测.比较各组间肺组织病理改变,湿重(W)/干重(D)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、丙二醛(MDA)、超氧化物歧化酶(SOD)含量,裂解型Caspase-3、HMGB1、TLR4、NF-κB表达水平的差异.结果 模型组小鼠肺组织出现肺损伤的病理改变,W/D、TNF-α、IL-1β、IL-6、MDA的含量及裂解型Caspase-3、HMGB1、TLR4、NF-κB的表达水平均高于对照组,SOD的含量低于对照组(P＜0.05).不同剂量血必净组小鼠肺损伤的病理改变减轻,W/D、TNF-α、IL-1β、IL-6、MDA 的含量及裂解型 Caspase-3、HMGB1、TLR4、NF-κB 的表达水平低于模型组,SOD的含量高于模型组(P＜0.05).HMGB1+高剂量血必净组小鼠肺损伤的病理改变加重,W/D、TNF-α、IL-1β、IL-6、MDA的含量及裂解型Caspase-3、HMGB1、TLR4、NF-κB的表达水平均高于NC+高剂量血必净组,SOD的含量低于NC+高剂量血必净组(P＜0.05).结论 血必净注射液对脓毒症小鼠肺损伤具有保护作用,并减轻炎症反应、氧化应激、细胞凋亡,其相关的分子机制为抑制HMGB1/TLR4/NF-κB通路.
Abstract
Objective To investigate the protective effect of Xuebijing injection on lung injury in septic mice by reg-ulating the high mobility group protein box 1(HMGB1)/Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)pathway.Methods Male C57BL/6 mice were randomly divided into control,model,low,medium and high dose of Xuebijing groups,negative control(NC)group,NC+model group,NC+high-dose of Xuebijing group,and HMGB1+high-dose of Xuebijing group.A sepsis-induced lung injury model was established using cecal perforation.Before mod-eling,the mice were intravenously injected with NC lentivirus or HMGB1 lentivirus via the tail vein.On the day of mod-eling,the mice were intraperitoneally injected with Xuebijing injection(5,10 and 15 mL/kg)twice a day for three con-secutive days.Samples were collected and detected 24 h after the last administration.The pathological changes of lung tissue,wet weight(W)/dry weight(D),tumor necrosis factor-α(TNF-α),interleukin-1 β(IL-1β),interleukin-6(IL-6),malondialdehyde(MDA)and superoxide dismutase(SOD)contents,the expression levels of Cleaved caspase-3,HMGB1,TLR4 and NF-κB were compared among all groups.Results Pathological changes of lung injury were ob-served in the model group,with increases in the levels of W/D,TNF-α,IL-1β,IL-6,MDA,and expression of Cleaved caspase-3,HMGB1,TLR4,NF-κB,and decreased SOD content compared with the control group(P＜0.05).In Xuebijing treatment groups,pathological changes in lung injury were alleviated,with lower levels of W/D,TNF-α,IL-1β,IL-6,MDA and expression of Cleaved caspase-3,HMGB1,TLR4,NF-κB,and higher SOD content than the model group(P＜0.05).The HMGB1+high-dose of Xuebijing group showed aggravated lung injury pathology,with higher levels of W/D,TNF-α,IL-1β,IL-6 and MDA,and expression of Cleaved caspase-3,HMGB1,TLR4 and NF-κB,and lower SOD content than the NC+high-dose of Xuebijing group(P＜0.05).Conclusions Xuebijing injection has protective effect on lung injury in septic mice and alleviates inflammation,oxidative stress and apoptosis.The molec-ular mechanism related to this effect is the inhibition of HMGB1/TLR4/NF-κB pathway.
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