硫化氢通过P物质介导促进小鼠肺组织的神经源性炎性反应  

Hydrogen Sulfide Induces the Neurogenic Inflammatory Reaction of Lung Tissues in Mice Mediated by P Substance

作　　者：赵倩[1] 王长谦[1] 乔建瓯[2] 张绘莉[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院心内科,上海200011 [2]上海交通大学医学院附属第九人民医院呼吸内科,上海200011

出　　处：《中国临床医学》2015年第3期262-267,共6页Chinese Journal of Clinical Medicine

基　　金：上海市自然科学基金(编号:13ZR1424200)

摘　　要：目的:探讨硫化氢对小鼠肺组织神经源性炎性反应的影响及其作用机制。方法:饲养8周龄的雄性Balb/c小鼠,分别予以CP-96345(2.5 mg/kg,腹腔注射)、辣椒素(50 mg/kg,皮下注射)或capsazepine(15 mg/kg,皮下注射)预处理,然后随机腹腔注射0.9%氯化钠溶液或硫氢化钠(NaHS;10 mg/kg),即分为对照组、NaHS组、CP-96345+NaHS组、辣椒素+NaHS组和capsazepine+NaHS组。用药1 h后处死小鼠,采用酶联免疫吸附试验(ELISA)法检测血浆P物质以及肺组织匀浆中肿瘤坏死因子-α(tumor necrosis factor-alpha,TNF-α)和白细胞介素-1β(interleukine-1 beta,IL-1β)水平,并用四甲基联苯胺反应液测定肺组织匀浆中髓过氧化物酶(myeloperoxidase,MPO)活性,通过HE染色法观察肺组织的病理学变化。另选8周龄、雄性、前速激肽原A(pre-pro-totchykinin A,PPT-A)基因敲除小鼠(PPT-A-/-)和相同遗传背景的同龄、野生型Balb/c小鼠(PPTA+/+),分别随机腹腔注射0.9%氯化钠液或NaHS(2.5 mg/kg),1 h后处死小鼠,检测肺组织匀浆中MPO活性、TNF-α和IL-1β水平,并观察肺组织的病理学变化。结果:NaHS可显著升高小鼠血浆P物质浓度(P<0.05),提高肺组织匀浆中的MPO活性以及TNF-α和IL-1β水平(均P<0.05),导致肺组织炎性损伤。敲除小鼠编码P物质的PPT-A基因或者予以CP-96345、辣椒素或capsazepine预处理阻断P物质信号通路后,NaHS不能显著升高肺组织中MPO活性以及TNF-α和IL-1β水平,与单纯NaHS处理组相比差异有统计学意义(均P<0.05);而且NaHS所致的肺组织损伤也明显减轻。结论:硫化氢可能通过刺激神经末梢纤维释放P物质,促进肺组织发生神经源性炎性反应,最终导致肺组织损伤。Objective：To investigate the effect of hydrogen sulfide on the neurogenic inflammation in mouse lung tissues,and its mechanism.Methods：Eight-week old,male Balb/c mice were randomly divided into the following groups：control group,NaHS group,CP-96345 ＋ NaHS group、capsaicin＋ NaHS group and capsazepine＋ NaHS group.The mice were pretreated with vehicle,CP-96345（2.5 mg/kg,i.p.）,capsaicin（50 mg/kg,s.c.）or capsazepine（15 mg/kg,s.c.）and then received NaHS（10 mg/kg,i.p.）or 0.9% NaCl solution.One hour after intervention,the mice were sacrificed,then the concentration of substance P in plasma and the levels of tumor necrosis factor-alpha（TNF-α）and interleukine-1 beta（IL-1β）in lung homogenate were measured by ELISA assay.Myeloperoxidase（MPO）activity in lung tissues was detected by tetramethyl benzidine（TMB）.The pathological changes of lung tissues were stained with hematoxylin and eosin.On the other hand,the preprotachykinin-A（PPT-A）gene knockout mice（eight-week old,female,PPT-A-/-）and their corresponding wild-type Balb/c mice（eight-week old,female,PPT-A＋/＋）were randomly given 0.9% NaCl solution or NaHS（10 mg/kg,i.p.）.One hour after intervention,the mice were sacrificed,then MPO activity,TNF-αand IL-1βlevels in lung tissues as well as the pathological changes of lung tissues were examined.Results：Hydrogen sulfide donor drug NaHS elevated significantly plasma substance P concentration（P〈0.05）,lung MPO activity as well as lung TNF-αand IL-1βlevels（all P〈0.05）,resulting in lung inflammatory injury.The knockout of PPT-A gene,or pretreatment with CP-96345,capsaicin or capsazepine abolished significantly the elevation of lung MPO activity,lung TNF-αand IL-1βlevels,as well as the lung injury induced by NaHS（all P〈0.05）.Conclusions：Hydrogen sulfide may induce lung inflammatory injury through stimulating the release of substance P in nerve endings.

关 键 词：硫化氢 炎性反应 P物质 肺损伤 

分 类 号：R563.1[医药卫生—呼吸系统]