清肺通络方通过胱天蛋白酶-1/Gasdermin D蛋白细胞焦亡途径对支原体肺炎小鼠的干预机制

目的:探讨清肺通络方通过胱天蛋白酶-1(Caspase-1)/Gasdermin D(GSDMD)蛋白细胞焦亡途径对支原体肺炎小鼠干预机制。方法:将30只BALB/c小鼠随机分为空白对照组、模型组、清肺通络方组，每组10只。采用肺炎支原体菌株滴鼻法建立支原体肺炎小鼠模型。造模同时予灌胃治疗，连续治疗3 d,实验第4天处死各组小鼠。PCR检测小鼠肺组织MP-DNA含量，HE染色观察小鼠肺组织病理改变，Western blot检测胱天蛋白酶-1前体(pro-Caspase-1)、Caspase-1、GSDMD、Gasdermin D氮端活性结构域多肽(GSDMD-N)表达，ELISA检测肺泡灌洗液中白介素-1β(IL-1β)、白介素-18(IL-18)含量，生化法检测肺组织细胞乳酸脱氢酶(LDH)和Na⁺-K⁺-ATP酶活性。结果:镜下观察模型组小鼠肺间质出现炎性细胞浸润、渗出明显。与空白对照组比较，模型组小鼠pro-Caspase-1、Caspase-1、GSDMD、GSDMD-N表达及IL-1β、IL-18含量均升高，LDH活性上升，Na

Intervention effects of Qingfei Tongluo recipe on Caspase-1/GSDMD cell death pathway in mice with mycoplasma pneumonia

Objective:To investigate the intervention effect of Qingfei Tongluo recipe on mycoplasma pneumonia(MP)mice through Caspase-1/GSDMD cell pyrodeath pathway.Methods:30 BALB/c mice were randomly assigned to an blank control group,model group and Qingfei Tongluo recipe group,with 10 mice in each group.The mouse model of mycoplasma pneumoniae was established by nasal drip of mycoplasma pneumoniae strains.The mice in each group were killed on the fourth day of the experiment.The content of MP-DNA in mouse lung tissue was detected by PCR.The pathological changes of lung tissue in each group were observed by HE staining.The expression of pro-Caspase-1,Caspase-1,GSDMD and GSDMD-N was detected by Western blot.The content of IL-1β and IL-18 in alveolar lavage fluid was detected by ELISA.The activity of LDH and Na⁺-K⁺-ATPase in lung tissue cells were detected by biochemical method.Results:The infiltration and exudation of inflammatory cells in lung interstitium of model group were observed under microscope.Compared with blank control group,the expressions of pro-Caspase-1,Caspase-1,GSDMD,GSDMD-N and the contents of IL-1β and IL-18 were increased in model group,while the activity of LDH was increased and the activity of Na⁺-K⁺-ATPase was decreased(P<0.05).Compared with model group,the infiltration of lung interstitial inflammatory cells in Qingfei Tonglu recipe group was significantly reduced,the expressions of Caspase-1,GSDMD,GSDMD-N and the contents of IL-1β and IL-18 in lung tissue were decreased(P<0.05),the activity of LDH was decreased,and the activity of Na⁺-K⁺-ATPase was increased(P<0.05).There was no significant difference in pro-Caspase-1 expression between the two groups(P>0.05).Conclusion:Qingfei Tongluo recipe alleviates pyrodeath and lung tissue inflammation in mice by inhibiting Caspase-1/GSDMD pathway.