摘要
目的 本研究旨在探讨血必净注射液(简称血必净)对脓毒症相关急性呼吸窘迫综合征(acute respiratory distress syndrome,ARDS)的保护作用机制.方法 ①动物实验:将100只小鼠随机(随机数字法)分为4组,包括假手术(Sham)组、盲肠结扎穿孔术(CLP)组、CLP+低剂量血必净(L-XBJ)组、CLP+高剂量血必净(H-XBJ)组,测定小鼠的存活率、肺组织学改变、肺湿/干(W/D)比、支气管肺泡灌洗液(bronchoalveolar lavage fluids,BALF)的细胞计数和蛋白浓度、血清中炎症因子水平、氧化应激指标、细胞凋亡、HIF-1α/p38 MAPK/NF-κB信号通路关键蛋白;②细胞实验:体外培养小鼠肺泡巨噬细胞(MH-S),分为6组,包括对照(Con)组、脂多糖(LPS)组、LPS+L-XBJ 组、LPS+H-XBJ 组、LPS+H-XBJ+二甲基草酰甘氨酸(DMOG,HIF-1 α激动剂)组、LPS+H-XBJ+二甲氧基雌二醇(2ME2,H1F-1 α抑制剂)组,检测血必净对炎症因子、氧化应激、细胞凋亡的影响及其与HIF-1α/p38 MAPK/NF-κB信号通路的关系.结果 血必净能够提高脓毒症相关ARDS小鼠的存活率,减轻肺组织损伤[肺损伤评分:CLP组(8.778±0.588)、CLP+L-XBJ 组(5.833±0.310)、CLP+H-XBJ 组(4.750±0.246)],降低肺 W/D 比,减少肺炎细胞浸润与蛋白渗出(均P＜0.05).此外,血必净还可以减少LPS诱导的MH-S细胞和CLP诱导的脓毒症相关ARDS小鼠炎症因子(TNF-α,IL-1 β和IL-6)的表达,降低体内外细胞内活性氧(reactive oxygen species,ROS)的积累,丙二醛(malondialdehyde,MDA)的形成,超氧化物歧化酶(superoxide dismutase,SOD)的消耗和细胞凋亡(均P＜0.05).机制研究进一步阐明了血必净通过HIF-1α/p38 MAPK/NF-κB信号通路对炎症、氧化应激和细胞凋亡的影响.结论 血必净可通过抑制HIF-1α/p38 MAPK/NF-κB信号通路,发挥抗炎、抗氧化和抗凋亡作用,从而对脓毒症相关ARDS产生保护作用.
Abstract
Objective To explore the protective mechanism of Xuebijing injection(referred to as Xuebijing)on sepsis-associated acute respiratory distress syndrome(ARDS).Methods ① Animal experiments:100 mice were randomly(random number)divided into 4 groups,including sham operation(Sham)group,cecal ligation and puncture(CLP)group,CLP+low-dose Xuebijing(L-XBJ)group,and CLP+high-dose Xuebijing(H-XBJ)group.The survival rate,lung histological changes,lung wet/dry(W/D)ratio,cell count and protein concentration in bronchoalveolar lavage fluids(BALF),inflammatory factors levels in serum,oxidative stress indicators,cell apoptosis,and key proteins of HIF-1α/p38 MAPK/NF-κB signaling pathway were measured.(2)Cell experiments:Mouse alveolar macrophages(MH-S)were cultured in vitro and divided into 6 groups,including control(Con)group,lipopolysaccharide(LPS)group,LPS+L-XBJ group,and LPS+H-XBJ group,LPS+H-XBJ+dimethyloxallyl glycine(DMOG,HIF-1α activator)group,LPS+H-XBJ+2-methoxyestradiol(2ME2,HIF-1α inhibitor)group.The effects of Xuebijing on inflammatory factors,oxidative stress,and cell apoptosis and their relationship with HIF-1α/p38 MAPK/NF-κB signaling pathway were detected.Results Xuebijing increased the survival rate of mice with sepsis-associated ARDS,relieved lung tissue damage[lung injury score:CLP group(8.778±0.588),CLP+L-XBJ group(5.833±0.310),and CLP+H-XBJ group(4.750±0.246)],alleviated lung W/D ratio,and decreased pneumonia cell infiltration and protein exudation(all P＜0.05).Additionally,Xuebijing treatment also diminished the expression of inflammatory factors(TNF-α,IL-1β,and IL-6),intracellular reactive oxygen species(ROS)accumulation,malondialdehyde(MDA)formation,superoxide dismutase(SOD)depletion,and cell apoptosis in LPS-induced MH-S cells and CLP-induced sepsis-associated ARDS mice(all P＜0.05).Furthermore,mechanistic investigation further clarified the effects of Xuebijing on inflammation,oxidative stress,and cell apoptosis through the HIF-1α/p38 MAPK/NF-κB signaling pathway.Conclusions Xuebijing can exert anti-inflammatory,anti-oxidative,and anti-apoptotic effects by suppressing the HIF-1α/p38 MAPK/NF-κB signaling pathway,thereby conferring protection against sepsis-associated ARDS.
基金项目
国家自然科学基金(82172182)
国家自然科学基金(82102311)
江苏省自然科学基金面上项目(BK20211136)
江苏省徐州市科技计划项目(KC22136)
徐州医科大学附属医院科技发展基金项目(XYFY202232)
北京协和医学基金-睿E(睿意)急诊医学研究专项基金项目(22222012001)
NETL
NSTL
万方数据