SS-31抑制O_(3)介导的小鼠气道高反应和黏液高分泌

SS-31 inhibits O_(3)-mediated airway hyperresponsiveness andmucus hypersecretion in mice

目的探讨线粒体靶向抗氧化肽SS-31是否能够抑制臭氧(O_(3))介导的小鼠肺部气道高反应性和黏液高分泌。方法将8周C57BL/6健康小鼠随机分为磷酸缓冲盐溶液组(PBS组)+空气组(Air组)、SS-31+Air组、PBS+O_(3)组、SS-31+O_(3)组。O_(3)暴露前1 h,C57BL/6小鼠接受腹腔注射SS-31(10 mg/kg),小鼠单次暴露于O_(3)环境,浓度5.01×10^(-6) mol/m 3,时间持续3 h。24 h后,测定小鼠的气道高反应(AHR),支气管肺泡灌洗液(BALF)细胞计数,肺组织过碘酸雪夫染色(PAS)、肺组织丙二醛(MDA)、白介素(IL)-1β、IL-6、IL-18、单核细胞趋化因子-1(MCP-1)及黏液分子(MUC5B)的mRNA表达水平,Western blot检测NOD样受体热蛋白结构域相关蛋白3(NLRP3)、半胱氨酸蛋白酶-1前体(pro-Caspase 1)/血清半胱氨酸蛋白酶-1剪切体Caspase 1(p20)、Gasdermin家族蛋白D(GSDMD)/Gasdermin家族蛋白D剪切体(Cleaved GSDMD)蛋白水平。结果O_(3)暴露引起小鼠气道高反应和黏液分泌增加。而SS-31能够抑制O_(3)介导的气道高反应和黏液分泌,降低O_(3)诱导的氧化应激水平和炎症因子mRNA水平表达,下调NLRP3表达水平、Caspase 1和GSDMD活化形式。结论SS-31通过抑制NLRP3/Caspase 1/GSDMD信号通路以抑制O_(3)介导的小鼠气道高反应和黏液高分泌。

Objective To investigate whether Mitochondria-targeted antioxidant peptide SS-31 can inhibit the ozone(O_(3))-induced mice lung airway hyperresponsiveness and mucus hypersecretion.Methods Eight-week C57BL/6 mice were randomized into four groups,including phosphate buffer saline(PBS)+Air group,SS-31+Air group,PBS+O_(3)group and SS-31+O_(3)group.C57BL/6 mice were injected intraperitoneally with SS-31(10 mg/kg)one hour before ozone exposure,and then single-exposed to ozone at a concentration of 5.01×10^(-6) mol/m^(3) for 3 hours.After 24 hours,airway hyperresponsiveness(AHR)and bronchoalveolar lavage fluid(BALF)cells numbers were measured.Lung tissue schiff periodic acid shiff(PAS)staining,malondialdehyde(MDA),inflammatory factors(interleukin,IL)-1β,IL-6,IL-18 and monocyte chemoattractant protein-1(MCP-1))and mucin factor(MUC5B)were detected,and the protein expression levels of NOD-like receptor thermal protein domain associated protein 3(NLRP3),pro-Caspase 1/Caspase 1(p20),Gasdermin D(GSDMD)and Cleaved GSDMD were determined by Western blot.Results O_(3)exposure caused both mice lung airway hyperresponsiveness and mucus hypersecretion.However,SS-31 could inhibit the O_(3)-induced airway hyperresponsiveness and mucus secretion,reduce the levels of oxidative stress and inflammatory factor mRNA expression,and downregulate the protein expression level of NLRP3 and the activated forms of Caspase 1 and GSDMD.Conclusion SS-31 could suppress O_(3)-induced mice airway hyperresponsiveness and mucus hypersecretion by inhibiting the NLRP3/Caspase 1/GSDMD signaling pathway.