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目的观察卵白蛋白(OVA)诱导的大鼠急性支气管哮喘(简称哮喘)模型,内源性硫化氢(H_2S)生成的变化以及应用外源性硫氢化钠(NaHS,H_2S 供体)处理对哮喘大鼠的影响,探讨气体信号分子 H_2S 在哮喘发病中的作用。方法 24只健康 SD 大鼠按随机数字表法分为正常对照组、哮喘组和 NaHS 干预组,每组8只。致敏后28 d 测定所有大鼠肺功能并观察大鼠支气管周围炎性细胞浸润程度并进行评分;采用敏感硫电极测定血浆及肺组织 H_2S 的生成量;采用酶促反应法测定大鼠肺组织匀浆中胱硫醚-γ-裂解酶(CSE)活性;用 Western blot 法测定大鼠肺组织中 CSE 蛋白含量(每组3只)。结果哮喘组大鼠呼气峰流量(PEF)、血浆及肺组织中 H_2S 分别为(2.90±0.70)L/s、(10±3)、(4.9±1.3)μmol/L,对照组分别为(6.50±0.10)L/s、(54±10)、(24.1±8.0)μmol/L,NaHS 干预组大鼠分别为(5.70±0.50)L/s、(17±4)、(15.3±4.0)μmol/L,3组间比较差异有统计学意义(F 值分别为112.13、110.10、27.34,P 均<0.01);哮喘组大鼠肺组织匀浆每毫克蛋白中 CSE 活性和肺组织匀浆中 CSE 蛋白含量[用相对吸光度(A)值表示]分别为(1.00±0.10)nmol·min~(-1)·mg~(-1)、0.20±0.10,正常对照组分别为(1.80±0.10)nmol·min~(-1)·mg~(-1)、0.90±0.30,NaHS 干预组大鼠分别为(1.60±0.20)nmol·min~(-1)·mg~(-1)、1.10±0.20,3组间比较差异有统计学意义(F 值分别为79.39、12.28,P 均<0.05);光镜下支气管周围炎性细胞浸润程度评分[用中位数(四分位数)]表示,正常对照组为1(0～1)分,哮喘组为3(2～4)分,NaHS 干预组为1(1～2)分,3组间比较差异有统计学意义(H=16.93,P<0.01);哮喘组肺组织 H_2S 含量与 PEF 呈正相关(r=0.74,P<0.01);与光镜下支气管周围炎性细胞浸润程度评分呈负相关(r=-0.64,P<0.01)。结论内源性 H_2S 参与了大鼠急性哮喘发病过程,外源性 NaHS 可以减轻哮喘气道炎症,对哮喘急性发病起到保护作用。
Objective To observe the changes of endogenous hydrogen sulfide (H 2 S) induced by ovalbumin (OVA) -induced acute asthma in rats and the effects of exogenous sodium hydrosulfide (NaHS, H 2 S donor) on asthma Mouse, to explore the role of gas signaling molecule H_2S in the pathogenesis of asthma. Methods Twenty-four healthy SD rats were divided into normal control group, asthma group and NaHS intervention group by random number table method, with 8 rats in each group. The lung function of all the rats was measured at 28 days after sensitization and the infiltration of inflammatory cells around the bronchus was observed and scored. The production of H 2 S in plasma and lung tissue was measured by sensitive sulfur electrode. The activity of cystathionine-γ-lyase (CSE) in homogenate was determined. The contents of CSE protein in lung tissue were determined by Western blot (3 in each group). Results The peak expiratory flow (PEF) and H 2 S in plasma and lung tissue of asthmatic rats were (2.90 ± 0.70) L / s, (10 ± 3) and (4.9 ± 1.3) μmol / L, (5.70 ± 0.50) L / s, (17 ± 4), (15.3 ± 4.0) and (4.50 ± 0.10) L / s, μmol / L, the difference between the three groups was statistically significant (F values were 112.13,110.10,27.34, P all <0.01); CSE activity per milligram of protein in the lung tissue homogenate of asthma group and in lung tissue homogenate The contents of CSE protein [(relative absorbance (A)] were (1.00 ± 0.10) nmol · min -1 · mg -1 and 0.20 ± 0.10 respectively, and were 1.80 ± 0.10 in the normal control group respectively (1.60 ± 0.20) nmol · min ~ (-1) · mg -1, 1.10 ± 0.45, respectively. The levels of TNFα, 0.20,3, the difference was statistically significant (F = 79.39,12.28, P <0.05). The score of bronchial inflammatory cell infiltration in bronchus was expressed by median (quartile) (0 ~ 1) in normal control group, 3 (2 ~ 4) in asthma group and 1 (1 ~ 2) in NaHS intervention group.The difference between the three groups was statistically significant (H = 16.93, P <0.01); H_2S lung breath group content was positively correlated with PEF (r = 0.74, P <0.01); ambient light microscope bronchial inflammatory cell infiltration score was a negative correlation (r = -0.64, P <0.01). Conclusions Endogenous H 2 S is involved in the pathogenesis of acute asthma in rats. Exogenous NaHS can reduce airway inflammation in asthma and play a protective role in acute asthma.