摘要
目的 探讨氯气所致小鼠急性呼吸窘迫综合征(ARDS)中细胞外组蛋白对肝素酶的诱导激活作用。方法 将无特定病原体级C57BL/6雄性小鼠随机分为对照组、氯气损伤组、组蛋白损伤组、抗组蛋白抗体组和肝素酶抑制剂组,每组6只。采用动式吸入染毒法,对照组和组蛋白损伤组小鼠予洁净空气,其余3组小鼠予剂量为580.0 mg/m;的氯气一次性染毒30 min。组蛋白损伤组小鼠经尾静脉注射剂量为50 mg/kg体质量的小牛胸腺组蛋白。在染毒前1 h,采用尾静脉注射法,抗组蛋白抗体组小鼠予剂量为20 mg/kg体质量的抗组蛋白H4抗体,肝素酶抑制剂组小鼠予剂量为2 mg/kg体质量的肝素酶抑制剂OGT2115;其余3组小鼠注射等体积的0.9%氯化钠溶液。染毒后24 h,采集各组小鼠动脉血进行血气分析,取肺组织进行病理组织学检查,采用酶联免疫吸附实验检测肺组织中的肝素酶蛋白水平,采用乙酰肝素降解产物分析方法检测肝素酶的活力,采用免疫印迹法检测肝素酶原和活性肝素酶的蛋白相对表达水平。结果 与对照组比较,氯气损伤组和组蛋白损伤组小鼠、均出现呼吸困难,肺组织均出现弥漫性炎症损伤,动脉血氧合指数均下降(P值均<0.05),且肺组织中肝素酶的蛋白水平、酶活力和肝素酶原、活性肝素酶的蛋白相对表达水平均升高(P值均<0.05)。与氯气损伤组和组蛋白损伤组比较,抗组蛋白抗体组小鼠呼吸困难、低氧血症和急性肺损伤均减轻,小鼠肺组织中肝素酶蛋白水平和肝素酶原、活性肝素酶的蛋白相对表达水平均下降(P值均<0.05);且该组小鼠肺组织肝素酶活力低于氯气损伤组(P<0.05)。与氯气损伤组比较,肝素酶抑制剂组小鼠呼吸困难症状、低氧血症和急性肺损伤均减轻,小鼠肺组织中肝素酶活力和肝素酶原蛋白相对表达水平均下降(P值均<0.05)。结论 在氯气所致小鼠ARDS发生发展过程中,细胞外组蛋白可诱导激活肝素酶而促进肺损伤,抑制肝素酶诱导激活可减轻肺损伤。
Objective To investigate the induction and activation of heparinase by extracellular histones in acute respiratory distress syndrome(ARDS) induced by chlorine in mice.Methods The specific pathogen free adult male C57 BL/6 mice were randomly divided into control group, chlorine injured group, histone injured group, anti-histone antibody group and heparinase inhibitor group, with six mice in each group.The mice in the control group and histone injured group were exposed to clean air, and the mice in the other three groups were exposed to chlorine gas at a dose of 580.0 mg/m;for 30 minutes by systemic dynamic inhalation.Mice in the histone injured group were injected with 50 mg/kg body weight calf thymus histone by tail vein.One hour before exposure, mice in the anti-histone antibody group were pretreated with 20 mg/kg body weight anti-histone H4 antibody by tail vein injection, and mice in the heparinase inhibitor group were injected with 2 mg/kg body weight OGT2115(heparinase inhibitor). The other three groups were given equal volume of 0.9% sodium chloride solution by tail vein injection. After 24 hours of exposure, arterial blood was collected for blood gas analysis and the lung tissue was collected for histopathological examination. The protein level of heparinase in lung tissue were detected using enzyme-linked immunosorbent assay, and the activity of heparinase were detected by measuring the product of heparan degradation. The protein expression of pro-heparinase and active heparinase were detected by Western blotting.Results The dyspnea developed of mice in the chlorine injured group and histone injured group, diffuse inflammation occurred in lung tissue, the oxygenation index in arterial blood decreased(all P<0.05), and the protein level and activity of heparinase in lung tissue, as well as the relative expression of pro-heparinase and active heparinase were increased compared with the control group(all P<0.05). The dyspnea, hypoxemia and acute lung injury of mice in the anti-histone antibody group were alleviated, and the protein level of heparinase in lung tissue, as well as the relative expression levels of pro-heparinase and active heparinase were decreased(all P<0.05), compared with chlorine injury group and histone injury group.The dyspnea, hypoxemia and acute lung injury were alleviated in the heparinase inhibitor group, and the activity of heparinase and the relative expression of pro-heparinase in the lung tissue were decreased compared with the chlorine injury group(all P<0.05). Conclusion During the occurrence and development of chlorine-induced ARDS in mice, extracellular histones aggravate lung injury by inducing the expression and activation of heparinase. Acute lung injury can be alleviated by inhibiting the expression and activation of heparinase.
作者
张雁林
郭利霞
陈明
赵怡然
李晓
郑亦沐
关里
赵赞梅
关晓旭
李树强
赵金垣
ZHANG Yan-lin;GUO Li-xia;CHEN Ming;ZHAO Yi-ran;LI Xiao;ZHENG Yi-mu;GUAN Li;ZHAO Zan-mei;GUAN Xiao-xu;LI Shu-qiang;ZHAO Jin-yuan(Department of Occupational Medicine,Peking University Third Hospital,Beijing 100191,China)
出处
《中国职业医学》
CAS
北大核心
2021年第5期488-494,共7页
China Occupational Medicine
基金
国家自然科学基金面上项目(81773374)
北京市自然科学基金面上项目(7182179)
北京大学第三医院优秀留学回国人员科研启动基金(Y7551305)。
关键词
氯气
急性肺损伤
急性呼吸窘迫综合征
细胞外组蛋白
肝素酶
诱导
激活
炎症
Chlorine gas
Acute lung injury
Acute respiratory distress syndrome
Extracellular histones
Heparinase
Induction
Activation
Inflammation
