Reduning Attenuates LPS-Induced Human Unmilical Vein Endothelial Cells (HUVECs) Apoptosis Through PI3K-AKT Signaling Pathway

Abstract

The molecular mechanism of Reduning (RDN) in the treatment of sepsis was analyzed based on network pharmacology. The system pharmacology method was administered to search the active ingredients and targets of RDN, identify the sepsis-related genes, and determine the targets of RDN in the treatment of sepsis. Cytoscape was used to build a “drug component-target” network to screen key compounds. A protein-protein interaction (PPI) network was constructed using STRING, and core targets were revealed through topological analysis. 404 shared targets of RDN and sepsis were introduced into DAVID Bioinformatics Resources 6.8 for GO and KEGG enrichment analysis to predict their possible signaling pathways and explore their molecular mechanisms. GO enrichment analysis highlighted that they were largely related to protein phosphorylation, inflammatory reaction, and positive regulation of mitogen-activated protein kinase (MAPK) cascade. KEGG enrichment analysis outlined that they were enriched in PI3K-AKT signaling pathway, calcium signaling pathway, rhoptry-associated protein 1 (Rap1) signaling pathway, and advanced glycation end products and receptors for advanced glycation end products (AGE-RAGE) signaling pathway. Molecular biological validation results exposed that RDN could significantly improve the protein expression of p-AKT and p-PI3K, alleviate apoptosis-related proteins expression level and decrease apoptosis rate in LPS-induced HUVECs. In conclusion, it was illustrated that RDN could considerably constrain LPS-induced apoptosis by activating the PI3K-AKT signaling pathway, which advocated a basis for fundamental mechanism research and clinical application of RDN in the treatment of sepsis.

Feasibility and mechanism analysis of Reduning in the prevention of sepsis-induced pulmonary fibrosis

“…Reduning is a TCM injection composed of Artemisia annulosa, honeysuckle, and gardenia, which is recommended for the treatment of critical COVID-19 patients by "the Diagnosis and Treatment Protocol for ." Our previous studies confirmed that Reduning could improve sepsis-induced acute lung injury (SALI) by downregulating LPS-induced apoptosis of human umbilical vein endothelial cells (HUVECs) through the PI3K-AKT pathway and thereby improving vascular endothelial barrier function (Wang et al, 2022a). Existing studies have shown that, the basic treatment of Western medicine, supplemented with Reduning, could significantly improve the clinical symptoms, pulmonary function, and prognosis of patients with idiopathic fibrosis (Wang et al, 2020;Shi et al, 2022).…”

In silico high-throughput screening system for AKT1 activators with therapeutic applications in sepsis acute lung injury

“…Activation of the PI3K-AKT-mTOR pathway could alleviate the apoptosis of HUVECs ( Lin et al., 2020 ; Chen et al., 2021 ). In addition, our previous study revealed that various active components in Reduning, a traditional Chinese medicine, could alleviate LPS-induced apoptosis of HUVECs by targeting AKT1 and activating AKT phosphorylation ( Wang et al., 2022 ). The discovery of AKT dates back to the 1970s, when an oncogene sequence, named AKT8, was identified in murine leukemia viruses, and two homologous oncogenes of AKT8, named AKT1 and AKT2 (also known as PKKB αand PKKB β, respectively), were subsequently identified in human chromosomes ( Pastorino et al., 2005 ).…”

“…There are 3 isoforms of AKT-AKT1, AKT2, and AKT3-among which AKT1 is most closely connected to apoptosis (Green et al, 2013;Gong et al, 2017;Kim et al, 2020). Previous studies have confirmed that pyrotoxin could improve apoptosis of LPSinduced HUVECs by promoting AKT1 phosphorylation (Wang et al, 2022). Currently, the only activator targeting AKT is SC79, which binds to the pleckstrin homology (PH) domain of AKT at residue 25 (arginine) (Jo et al, 2012).…”

Reduning alleviates sepsis-induced acute lung injury by reducing apoptosis of pulmonary microvascular endothelial cells

“…For example, Tang et al stated that RDN can improve lung inflammation and oxidative stress by down-regulating the nuclear factor (NF)-kB pathway (15). Yang et al found that RDN ameliorated LPS-induced ALI through decreased phosphorylation of extracellular regulated protein kinase (ERK)1/2 to suppress the formation of neutrophil extracellular traps (NETs) (16). In our previous research, we found that the activation of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) pathway by RDN could reduce the lipopolysaccharide (LPS)induced apoptosis of human umbilical vein endothelial cells (HUVECs) (17).…”

“…found that RDN ameliorated LPS-induced ALI through decreased phosphorylation of extracellular regulated protein kinase (ERK)1/2 to suppress the formation of neutrophil extracellular traps (NETs) ( 16 ). In our previous research, we found that the activation of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) pathway by RDN could reduce the lipopolysaccharide (LPS)-induced apoptosis of human umbilical vein endothelial cells (HUVECs) ( 17 ). In this study, we further explored the underlining mechanisms by which RDN can inhibit apoptosis through the AKT pathway.…”