基于宏基因组探讨青蒿鳖甲汤对脓毒症急性肺损伤大鼠的影响

Effects of Qinghao Biejia Decoction in acute lung injury of sepsis rats based on metagenome

目的:基于宏基因组探讨青蒿鳖甲汤对肠缺血再灌注诱发脓毒症急性肺损伤微生物多样性及碳水化合物酶类(CAZy)功能谱的影响。方法:32只SPF级Wistar雄性大鼠随机分为假手术组、模型组、青蒿鳖甲汤组、地塞米松组。夹闭肠系膜上动脉建立肠缺血再灌注模型。HE染色观察大鼠肺、肠组织形态变化,免疫组化法检测大鼠肺、肠组织中MyD88的表达。采集结肠内容物,提取DNA,进行宏基因组分析。结果:与模型组比较,青蒿鳖甲汤组减轻细支气管及肺泡扩张充血,肺泡巨噬细胞浸润;显著抑制肺泡、胃黏膜MyD88信号表达(P<0.01,P<0.05),减轻炎症反应;宏基因组学分析表明青蒿鳖甲汤组能够有效恢复大鼠肠道菌群丰富度和多样性,提升GH2、GH3、GH13、GH43水平(P<0.05,P<0.01),并抑制β-葡萄糖醛酸酶表达(P<0.01)。结论:微生物群的变化有助于调节CAZy的丰度和β-葡萄糖醛酸酶活性,青蒿鳖甲汤可能作为酶抑制剂发挥抑制肠缺血再灌注过度炎症反应作用,从而减轻肠缺血再灌注引起的脓毒症肺损伤。

Objective:To investigate the effects of Qinghao Biejia Decoction(QHBJT) on microbial diversity and carbohydrate enzymes(CAZy) function spectrum of acute lung injury of sepsis induced by intestinal ischemia-reperfusion.Methods:Thirty-two rats were randomly divided into sham operation group,model group,QHBJT group and dexamethasone group.The superior mesenteric artery was clipped to establish an intestinal is chemia-reperfusion model.HE staining was used to observe the morphological changes of lung and intestinal tissues of the rats,and immunohistochemical method was used to detect the expression of MyD88 in lung and intestinal tissues.Colon contents were collected and DNA was extracted for metagenomic analysis.Results:Compared with the model group,QHBJT reduced bronchiole and alveolar dilatation and congestion,and alveolar macrophage infiltration;could significantly inhibit MyD88 signal expression(P<0.01,P<0.05) and reduce inflammatory response;Metagenomic analysis showed that QHBJT could effectively restore the richness and diversity of intestinal flora,increase the levels of GH2,GH3,GH13 and GH43(P<0.05,P<0.01),and inhibit the expression of β-glucuronase in rats(P<0.01).Conclusion:The changes of microflora can regulate the abundance of CAZy and the activity of β-glucuronase.QHBJT may play an enzyme inhibitor role in inhibiting the excessive inflammatory response of intestinal ischemia reperfusion,so as to reduce the intestinal ischemia reprefusion-induced lung injury of sepsis.