datasheet1_Shenling Baizhu Powder Inhibits RV-SA11-Induced Inflammation and Rotavirus Enteritis via TLR4/MyD88/NF-κB Signaling Pathway.pdf

Rotavirus enteritis (RVE) is a common acute intestinal infectious disease caused by rotavirus infection. It is an important cause of death in children younger than 5 years worldwide. Shenling baizhu powder (SBP), a classic traditional Chinese formulation, is one of the most popularly prescribed medicines for digestive diseases. Clinical studies have revealed the protective effects of SBP on RVE. However, the potential mechanism is still unclear. In this study, we aimed to evaluate the anti-rotavirus effect of SBP and its mechanism, focusing on the TLR4/MyD88/NF-κB signaling pathway. Our results demonstrated that, based on the inhibition of the virus-induced cytopathic effect in Caco-2 cells, the concentration for 50% of maximal effect (EC50) and selectivity index (SI) of SBP for RV-SA11 in the serum were 5.911% and 11.63, respectively. A total of 219 active compounds with oral bioavailability ≥30% and drug-likeness ≥ 0.18 were selected from the 10 ingredients present in the formulation of SBP, which acted on 471 potential targets. A total of 226 target genes of RVE were obtained from the GeneCards database. The protein-protein interaction (PPI) network showed that there was a close interaction between 44 common targets of SBP and RVE. The results of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that SBP acted on RVE through various inflammatory pathways and the intestinal immune network. Subsequently, we investigated the effect of SBP on TLR4/MyD88/NF-κB signaling pathway in vitro. After infection with RV- SA11, the expression of TLR4, MyD88, and NF-κB mRNA and protein increased significantly, which could be abolished by SBP treatment. In addition, the IL-1β, TNF-α, IL-6, and IFN-β levels increased markedly in Caco-2 cells infected with RV-SV11. Treatment with SBP partly reversed the changes of IL-1β, TNF-α, and IL-6, while further increased the level of IFN-β. In conclusion, our study revealed that SBP can significantly inhibit rotavirus replication and proliferation in vitro. The antiviral effect may be related to the regulation of the TLR4/MyD88/NF-κB signaling pathway, followed by the down regulation of inflammatory cytokines and up regulation of IFN-β induced by rotavirus.

轮状病毒肠炎（Rotavirus enteritis, RVE）是由轮状病毒感染引发的常见急性肠道传染病，是全球5岁以下儿童死亡的重要诱因之一。参苓白术散（Shenling baizhu powder, SBP）作为经典中药方剂，是临床治疗消化系统疾病最常用的药物之一。已有临床研究证实SBP对RVE具有保护作用，但其潜在作用机制仍未明确。
本研究旨在评估SBP的抗轮状病毒效应及其作用机制，重点聚焦TLR4/MyD88/NF-κB信号通路。研究结果显示，基于对Caco-2细胞中病毒诱导细胞病变效应的抑制作用，SBP在血清中针对RV-SA11的半数有效浓度（EC50）和选择性指数（SI）分别为5.911%和11.63。
研究团队从SBP的10味组方药材中，筛选得到219个口服生物利用度≥30%、类药性≥0.18的活性成分，这些成分共作用于471个潜在靶点。从GeneCards数据库中获取到226个RVE相关靶基因。蛋白质-蛋白质相互作用（PPI）网络分析显示，SBP与RVE的44个共同靶点之间存在紧密的相互关联。
基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析结果表明，SBP可通过多种炎症通路及肠道免疫网络发挥抗RVE作用。
随后本研究体外验证了SBP对TLR4/MyD88/NF-κB信号通路的调控效应：RV-SA11感染后，Caco-2细胞中TLR4、MyD88及NF-κB的mRNA与蛋白表达水平显著升高，而SBP干预可逆转这一变化。此外，RV-SV11感染的Caco-2细胞中IL-1β、TNF-α、IL-6及IFN-β水平均显著升高。SBP干预可部分逆转IL-1β、TNF-α及IL-6的异常升高，却进一步提升了IFN-β的表达水平。
综上，本研究证实SBP可在体外显著抑制轮状病毒的复制与增殖，其抗病毒效应可能通过调控TLR4/MyD88/NF-κB信号通路，进而下调轮状病毒诱导的炎症细胞因子表达，并上调IFN-β水平实现。