Jieduquyuziyin prescription (JP) has been used to treat lupus nephritis (LN) and its effectiveness in the treatment of LN has been clinically proven, but the underlying mechanisms have yet to be completely understood. This aim of this study was to clarify the efficacy of JP on the epithelial-mesenchymal transition (EMT) of renal tubular epithelial cells and the molecular mechanisms of JP in MRL/lpr mice. In vivo, we observed the therapeutic actions of JP in MRL/lpr mice as well as its antifibrosis effect and potential mechanism. In vitro, we evaluated the role of JP in EMT and its possible mechanism through the EMT of human renal proximal tubular epithelial cells (HK-2) induced by transforming growth factor-beta 1 (TGF-β1) and M2c macrophages. HK-2 cells were treated with JP-treated serum, and MRL/lpr mice were treated by JP for 8 weeks. The results showed that JP alleviated disease activity, improved renal function, decreased proteinuria, and improved renal injury and fibrosis in MRL/lpr mice. Furthermore, JP suppressed the activation of the TGF-β1/Smad2/3 signaling pathway, upregulated the E-cadherin levels, and downregulated the Vimentin and mesenchymal α-smooth muscle actin (α-SMA) levels in the kidney of MRL/lpr mice. JP was further found to prevent the TGF-β1 and M2c macrophages-induced EMT of HK-2 cells. Collectively, JP could alleviate the disease activity of MRL/lpr mice, improve renal function, and attenuate renal fibrosis, and its underlying mechanisms may be related to the inhibition of EMT and TGF-β1/Smad2/3 signaling pathway.

Interleukin-1receptor-associated kinase 1 (IRAK1) plays a critical role in systemic lupus erythematosus (SLE). It was reported that SLE was associated with an inflammatory response mediated by defective immune tolerance, including overproduction of autoantibodies, chronic inflammation, and organ damage. Previous reports stated paeoniflorin (PF) had an immunosuppressive effect. The purpose of this study was to determine the anti-inflammatory effect of PF in SLE and its underlying mechanisms. Followed by induced with lipopolysaccharide (LPS), the splenocytes and the isolated CD4⁺ T lymphocytes of MRL/lpr mice were divided into three groups: control group, LPS group, and LPS+PF group, respectively. MRL/MP mice were used as the control group (treated with distilled water). The MRL/lpr mice were randomly divided into three groups: the model group (treated with distilled water), the prednisone group, and the PF group. The MRL/lpr mice were treated with prednisone acetate (5mg/kg) and PF (25, 50, and 75mg/kg) for eight weeks. Subsequently, ELISA, qRT-PCR, western blotting, HE, and Masson staining were performed to detect various indicators. The results of Cell Counting Kit-8 (CCK-8) showed that 10μg/mL of LPS had the optimum effect on cell viability, and 50μmol/L of PF had no obvious cytotoxicity to LPS-treated cells. PF reduced the expression level of IRAK1-nuclearfactor-κB (NF-κB) and its downstream inflammatory cytokines in the splenocytes and CD4⁺ T lymphocytes of MRL/lpr mice stimulated by LPS, especially in the latter. The serum antibody contents in the PF group mice were reduced, and the kidney damage was also alleviated accordingly. Moreover, the IRAK1/inhibitor of the nuclear factor-κB kinase (IKK)/NF-κB inhibitor (IκB)/NF-κB pathways was found to be involved in the anti-inflammation effect of PF in the kidney and spleen. In conclusion, it is thought that PF may have the potential to be used as a therapeutic agent to reduce the inflammatory activity of SLE. Inhibition of the IRAK1-NF-κB pathway may help formulate novel therapeutic tactics for SLE.

Bushen Jianpi Quyu Formula (BSJPQYF), an experienced formula, has been used to treat aplastic anemia (AA) more than three decades. To determinate the effect of BSJPQYF on AA, we constructed an immune-mediated AA mouse model. All mice were divided into four groups: control, model, low dose (0.85g/mL), and high dose (1.7g/mL BSJPQYF) group. They were administered with different concentrations of BSJPQYF or normal saline for 14 days. Besides, components of BSJPQYF were analyzed by electrospray ionization and mass spectrometry (ESI-MS). Subsequently, mouse peripheral blood and femurs were collected, and bone marrow mesenchymal stem cells (BMSCs) were isolated by fluorescence-activated cell sorting (FACS). Among them, tumor necrosis factor-α (TNF-α), transforming growth factor-β (TGF-β), and interferon-γ (IFN-γ) were measured by ELISA assay, PI3K, AKT, p-AKT, NF-κB, p-NF-κB, TNF-α, and cleaved caspase-3 proteins were detected by western blot. Compared with standard compounds, we identified three compounds of BSJPQYF, namely, icariin, kaempferol and tanshinone iia, as potentially effective compounds for the treatment of AA. Through an in vivo study, we found the administration of BSJPQYF in high dose for 14 days could significantly increase peripheral blood count and bone marrow (BM) cells, meanwhile decrease TNF-α, TGF-β, and IFN-γ levels. Besides, it could suppress the protein expression of PI3K and the phosphorylation of AKT and NF-κB to restrict the protein expression of TNF-α, eventually reduce the protein expression of cleaved caspase-3. This study demonstrated the therapeutic effects of BSJPQYF in AA, which could alleviate myelosuppression through inhibiting the expression of the PI3K/AKT/NF-κB signaling pathway.

Background: Systemic lupus erythematosus (SLE) is an autoimmune inflammatory connective tissue disease involving multiple organs. As one of the serious complications of SLE, sepsis (SEP) has a high risk of death. Here, the goal of this study was to identify vulnerable biomarkers that could be used to diagnose SLE and SEP. Methods: We used the Limma R software tool and the Gene Expression Omnibus (GEO) database to find differentially expressed genes (DEGs) in SLE. Additionally, genes associated with oxidative stress and immune system function were chosen from the MSigDB database and the Genecard database, respectively. Weighted gene Coexpression network analysis (WGCNA) was used to identify the important module genes associated with SEP. With the help of WGCNA, machine learning, and logistic regression, immunological and oxidative stress-related hub genes were discovered and validated by an external validation set. The analysis was put to the test using consensus clustering. Immune cell infiltration was investigated in SLE and SEP patients. Results: We obtained 957 genes from the GSE6163 dataset and 2559 genes from the significant module of WGCNA, which yielded 46 genes after taking intersection with immune and oxidative stress-related genes. According to the enrichment analysis's findings, the two diseases share a lot of similar immunological and inflammation-related pathways. Machine learning was utilized to pick 11 hub genes, and ROC was employed to evaluate the diagnostic effectiveness. Furthermore, the expression profiles of the hub genes revealed by logistic regression modeling have a significant diagnostic value. Moreover, consensus clustering revealed a favorable correlation between the severity of immunological and oxidative stress and disease activity in SLE and SEP. Analysis of immune infiltration revealed a more consistent immune cell infiltration behavior between SLE and SEP. Conclusion: In this study, the expression of 11 potential hub genes, including TLR2, IL1RN, IRF9, ISG20, TXK, SH2D1A, IL7R, CD28, ITK, CD3E, and CCR7, were thoroughly analyzed using bioinformatics. An efficient logistic regression model was created, and it was possible to identify a correlation between the progression of SLE and SEP disease and the expression of immunological and oxidative stress by consensus clustering. In addition, there is a similar immunoinvasive behavior between the two diseases. It is helpful to identify the biological markers with potential diagnostic value.