COVID-19 is often characterized by dysregulated inflammatory and immune responses. It has been shown that the Traditional Chinese Medicine formulation Qing-Fei-Pai-Du decoction (QFPDD) is effective in the treatment of the disease, especially for patients in the early stage. Our network pharmacology analyses indicated that many inflammation and immune-related molecules were the targets of the active components of QFPDD, which propelled us to examine the effects of the decoction on inflammation. We found in the present study that QFPDD effectively alleviated dextran sulfate sodium-induced intestinal inflammation in mice. It inhibited the production of pro-inflammatory cytokines IL-6 and TNFα, and promoted the expression of anti-inflammatory cytokine IL-10 by macrophagic cells. Further investigations found that QFPDD and one of its active components wogonoside markedly reduced LPS-stimulated phosphorylation of transcription factor ATF2, an important regulator of multiple cytokines expression. Our data revealed that both QFPDD and wogonoside decreased the half-life of ATF2 and promoted its proteasomal degradation. Of note, QFPDD and wogonoside down-regulated deubiquitinating enzyme USP14 along with inducing ATF2 degradation. Inhibition of USP14 with the small molecular inhibitor IU1 also led to the decrease of ATF2 in the cells, indicating that QFPDD and wogonoside may act through regulating USP14 to promote ATF2 degradation. To further assess the importance of ubiquitination in regulating ATF2, we generated mice that were intestinal-specific KLHL5 deficiency, a CUL3-interacting protein participating in substrate recognition of E3s. In these mice, QFPDD mitigated inflammatory reaction in the spleen, but not intestinal inflammation, suggesting CUL3-KLHL5 may function as an E3 for ATF2 degradation.

中文翻译：

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清肺排毒汤与汉黄芩苷通过下调USP14促进活化转录因子2降解发挥抗炎作用

COVID-19 通常以炎症和免疫反应失调为特征。研究表明，中药清肺排毒汤（QFPDD）对该病的治疗效果显着，尤其是对早期患者。我们的网络药理学分析表明，许多炎症和免疫相关分子是 QFPDD 活性成分的靶点，这促使我们检查汤剂对炎症的影响。我们在本研究中发现 QFPDD 有效减轻了葡聚糖硫酸钠诱导的小鼠肠道炎症。抑制促炎细胞因子IL-6和TNFα的产生，促进巨噬细胞表达抗炎细胞因子IL-10。进一步的研究发现，QFPDD 及其一种活性成分汉黄芩苷显着降低了 LPS 刺激的转录因子 ATF2 的磷酸化，转录因子 ATF2 是多种细胞因子表达的重要调节因子。我们的数据显示，QFPDD 和汉黄芩苷都降低了 ATF2 的半衰期并促进了其蛋白酶体降解。值得注意的是，QFPDD 和汉黄芩苷下调去泛素化酶 USP14，同时诱导 ATF2 降解。小分子抑制剂IU1对USP14的抑制也导致细胞中ATF2的减少，表明QFPDD和wogonoside可能通过调节USP14来促进ATF2降解。为了进一步评估泛素化在调节 ATF2 中的重要性，我们生成了肠道特异性 KLHL5 缺陷的小鼠，一种参与 E3 底物识别的 CUL3 相互作用蛋白。在这些小鼠中，QFPDD 减轻了脾脏的炎症反应，但不能减轻肠道炎症，这表明 CUL3-KLHL5 可能作为 E3 来降解 ATF2。