背景:双丹解毒汤(SJD)是由六种具有解热解毒功能的中药组成的配方,抗菌和抗炎作用,临床上用于治疗包括COVID-19在内的各种肺部炎症性疾病,但其作用的治疗物质基础及其分子机制尚不清楚。
目的:本研究试图确定SJD对LPS诱导的急性肺损伤(ALI)的治疗作用,以及研究其作用机制并评估其在临床上治愈炎症相关疾病的治疗潜力。
方法:我们通过气管滴注LPS建立了ALI模型,在SJD管理之后,收集小鼠支气管肺泡灌洗液和肺组织,检测其中炎症因子的表达。此外,我们通过免疫印迹评估了SJD对环磷酸鸟苷-磷酸腺苷合成酶-干扰素基因(cGAS-STING)和炎性体的刺激物的影响,和实时定量聚合酶链反应(RT-qPCR)。
结果:我们证明了SJD通过抑制支气管肺泡灌洗液(BALF)中的促炎细胞因子水平,可有效缓解LPS诱导的ALI,改善肺组织病理学水平和中性粒细胞数量,以及降低炎症因子相关基因的表达。重要的是,我们发现SJD可以抑制多种刺激驱动的cGAS-STING和炎性小体的激活。进一步的研究表明,在配方剂量下,SJD中的中草药对cGAS-STING途径和单独的炎性小体没有影响。通过增加这些草药的浓度,我们观察到对cGAS-STING途径和炎症小体的抑制作用,当六种草药混合使用时,效果最大,表明这些草药之间的协同作用在SJD的抗炎作用中起着至关重要的作用。
结论:我们的研究表明SJD对ALI具有良好的保护作用,其作用机制可能与6种中药抑制cGAS-STING和炎性小体异常激活的协同作用有关。这些结果有利于SJD在临床上的广泛应用以及从草药配方中开发ALI药物。
BACKGROUND: Shuangdan Jiedu Decoction (SJD) is a formula composed of six Chinese herbs with heat-removing and detoxifying, antibacterial, and anti-inflammatory effects, which is clinically used in the therapy of various inflammatory diseases of the lungs including COVID-19, but the therapeutic material basis of its action as well as its molecular mechanism are still unclear.
OBJECTIVE: The study attempted to determine the therapeutic effect of SJD on LPS-induced acute lung injury (ALI), as well as to investigate its mechanism of action and assess its therapeutic potential for the cure of inflammation-related diseases in the clinical setting.
METHODS: We established an ALI model by tracheal drip LPS, and after the administration of SJD, we collected the bronchoalveolar lavage fluid (BALF) and lung tissues of mice and examined the expression of inflammatory factors in them. In addition, we evaluated the effects of SJD on the cyclic guanosine monophosphate-adenosine monophosphate synthase -stimulator of interferon genes (cGAS-STING) and inflammasome by immunoblotting and real-time quantitative polymerase chain reaction (RT-qPCR).
RESULTS: We demonstrated that SJD was effective in alleviating LPS-induced ALI by suppressing the levels of pro-inflammatory cytokines in the BALF, improving the level of lung histopathology and the number of neutrophils, as well as decreasing the inflammatory factor-associated gene expression. Importantly, we found that SJD could inhibit multiple stimulus-driven activation of cGAS-STING and inflammasome. Further studies showed that the Chinese herbal medicines in SJD had no influence on the cGAS-STING pathway and inflammasome alone at the formulated dose. By increasing the concentration of these herbs, we observed inhibitory effects on the cGAS-STING pathway and inflammasome, and the effect exerted was maximal when the six herbs were combined, indicating that the synergistic effects among these herbs plays a crucial role in the anti-inflammatory effects of SJD.
CONCLUSIONS: Our research demonstrated that SJD has a favorable protective effect against ALI, and its mechanism of effect may be associated with the synergistic effect exerted between six Chinese medicines to inhibit the cGAS-STING and inflammasome abnormal activation. These results are favorable for the wide application of SJD in the clinic as well as for the development of drugs for ALI from herbal formulas.