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目的研究大环内酯类药联合糖皮质激素对哮喘大鼠肺组织的支气管肺泡灌洗液(bronchoalveolar lavagefluid,BALF)中嗜酸性粒细胞(eosinophil,EOS)凋亡相关蛋白cleaved caspase-9、Bak表达的影响。方法以卵白蛋白致敏制作大鼠哮喘模型,将大鼠分为5组:对照组、哮喘模型组、干预组A1(红霉素)、干预组A2(地塞米松)、干预组A3(红霉素+地塞米松)组,每组10只。并对其BALF中嗜酸性粒细胞进行计数,TUNEL法检测EOS凋亡;ELISA法测定BALF中IL-5、IL-8水平;Western blot检测EOS中cleaved caspase-9、Bak蛋白的表达。结果模型组BALF中细胞总数、EOS所占比例均显著高于对照组和干预组(P<0.01);凋亡率显著低于对照组和干预组(P<0.01)。对照组及干预组BALF中IL-5和IL-8浓度均低于模型组(P<0.01);干预组A3中两者浓度又分别低于干预组A1和干预组A2(P<0.01)。对照组、干预组分别与模型组相比,cleaved caspase-9、Bak蛋白表达的差异均有统计学意义(P<0.01);干预组A3分别与干预组A1、A2相比,cleaved caspase-9、Bak蛋白表达均有统计学差异(P<0.01)。结论大环内酯类药可协同糖皮质激素通过上调cleaved caspase-9、Bak的表达促进EOS细胞凋亡,从而发挥抗炎作用。
Objective To investigate the effect of macrolide combined with glucocorticoids on apoptosis of eosinophils (EOS) in bronchoalveolar lavage fluid (BALF) of asthmatic rats. The expressions of cleaved caspase-9, Bak The impact of expression. Methods The rat model of asthma was induced by ovalbumin. The rats were divided into 5 groups: control group, asthma model group, intervention group A1 (erythromycin), intervention group A2 (dexamethasone), intervention group A3 ADM + dexamethasone group, with 10 in each group. The number of eosinophils in BALF was counted. The apoptosis of EOS was detected by TUNEL method. The levels of IL-5 and IL-8 in BALF were measured by ELISA. The expression of cleaved caspase-9 and Bak in EOS were detected by Western blot. Results The total number of cells in the BALF and the percentage of EOS in the model group were significantly higher than those in the control group and the intervention group (P <0.01). The apoptosis rate was significantly lower in the model group than in the control group and the intervention group (P <0.01). The concentrations of IL-5 and IL-8 in BALF of the control group and the intervention group were lower than those of the model group (P <0.01). The concentration of A3 in the intervention group and the intervention group were lower than A1 and A2, respectively (P <0.01). Compared with the model group, the expression of cleaved caspase-9 and Bak protein in the control group and the intervention group were significantly different (P <0.01); Compared with the intervention group A1 and A2, the cleaved caspase-9 , Bak protein expression were statistically significant (P <0.01). Conclusion Macrolides can synergize with glucocorticoid to promote the apoptosis of EOS cells by up-regulating the expression of cleaved caspase-9 and Bak, thereby exerting anti-inflammatory effects.