Prednisone combined with Dihydroartemisinin attenuates systemic lupus erythematosus by regulating M1/M2 balance through the MAPK signaling pathway.

OBJECTIVE: Dihydroartemisinin (DHA) plays a very important role in various diseases. However, the precise involvement of DHA in systemic lupus erythematosus (SLE), relation to the equilibrium between M1 and M2 cells, remains uncertain. Therefore, we aimed to investigate the role of DHA in SLE and its effect on the M1/M2 cells balance. METHODS: SLE mice model was established by pristane induction. Flow cytometry was employed to measure the abundance of M1 and M2 cells within the peripheral blood of individuals diagnosed with SLE. The concentrations of various cytokines, namely TNF-α, IL-1ß, IL-4, IL-6, and IL-10, within the serum of SLE patients or SLE mice were assessed via ELISA. Immunofluorescence staining was utilized to detect the deposition of IgG and complement C3 in renal tissues of the mice. We conducted immunohistochemistry analysis to assess the expression levels of Collagen-I, a collagen protein, and α-SMA, a fibrosis marker protein, in the renal tissues of mice. Hematoxylin-eosin staining, Masson's trichrome staining, and Periodic acid Schiff staining were used to examine histological alterations. In this study, we employed qPCR and western blot techniques to assess the expression levels of key molecular markers, namely CD80 and CD86 for M1 cells, as well as CD206 and Arg-1 for M2 cells, within kidney tissue. Additionally, we investigated the involvement of the MAPK signaling pathway. The Venny 2.1 online software tool was employed to identify shared drug-disease targets, and subsequently, the Cytoscape 3.9.2 software was utilized to construct the "disease-target-ingredient" network diagram. Protein-protein interactions of the target proteins were analyzed using the String database, and the network proteins underwent enrichment analysis for Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathways. RESULTS: The results showed that an increase in M1 cells and a decrease in M2 cells within the peripheral blood of individuals diagnosed with SLE. Further analysis revealed that prednisone (PDN) combined with DHA can alleviate kidney damage and regulate the balance of M1 and M2 cells in both glomerular mesangial cells (GMC) and kidney. The MAPK signaling pathway was found to be involved in SLE kidney damage and M1/M2 balance in the kidney. Furthermore, PDN and/or DHA were found to inhibit the MAPK signaling pathway in GMC and kidney. CONCLUSION: We demonstrated that PDN combined with DHA attenuates SLE by regulating M1/M2 balance through MAPK signaling pathway. These findings propose that the combination of PDN and DHA could serve as a promising therapeutic strategy for SLE, as it has the potential to mitigate kidney damage and reinstate the equilibrium of M1 and M2 cells.

Shikonin combined with methotrexate regulate macrophage polarization to treat psoriasis.

This study aimed to investigate whether shikonin combined with methotrexate could inhibit psoriasis progression by regulating the polarization of macrophages through in vivo and in vitro experiments. Imiquimod was administrated to the exposed skin of BALB/c mice, and shikonin and methotrexate suspension were also given by gavage. The erythema, scales and thickness were scored for mice lesions in each group, and the total score was obtained by adding the above three scores, and calculated as psoriasis area and severity index (PASI) score. The skin lesion tissue from mice was isolated and used for hematoxylin-eosin staining and immunohistochemistry assay. Drug-containing serum was prepared and administrated into mouse macrophage RAW264.7 cells, followed by simulation of LPS. The levels of tumor necrosis factor-α (TNF-α), Interleukin (IL)-1ß, and IL-6 in cell supernatant were assessed using ELISA Kits and real-time PCR. In imiquimod-induced psoriasis mice, shikonin combined with methotrexate exerted protective effects by reducing erythema and PASI scores, decreasing backer score and epidermal thickness, and particularly regulating macrophage polarization. In LPS-stimulated RAW264.7 cells, shikonin combined with methotrexate regulated M1/M2 polarization and altered the levels of M1 markers. Shikonin combined with methotrexate inhibit psoriasis progression by regulating the polarization of macrophages, which may be useful in the treatment of psoriasis.

Dihydroartemisinin attenuated the symptoms of mice model of systemic lupus erythematosus by restoring the Treg/Th17 balance.

The Treg/Th17 imbalance is associated with the development of systemic lupus erythematosus (SLE). Dihydroartemisinin (DHA), a semi-synthetic derivative of artemisinin, is isolated from the traditional Chinese herb Artemisia annua Artemisia annua L. This study aims to evaluate the effects of DHA alone or in combination with prednisone in immunodeficiency of SLE. In vivo, the therapeutical effect of DHA alone or in combination with prednisone was assessed in the pristane-induced SLE mouse model. Then, the level of serum antibodies, creatinine (Cre), blood urea nitrogen (BUN), urine protein, kidney histopathology, interleukin (IL)-17, IL-6, transforming growth factor (TGF)-ß, the expression of RORγt and Foxp3, the percentages of Treg and Th17 in peripheral blood and spleen were assayed. In vitro, the mouse spleen lymphocytes were separated and treated with DHA alone or DHA in combination with prednisone. Then the percentages of Treg and Th17, the concentration of IL-17, IL-6, TGF-ß, and the expression of RORγt and Foxp3 were assayed. It was shown that DHA alone or in combination with prednisone treatment significantly alleviated the manifestations of pristane-induced SLE mice, suppressed inflammation and restored the Treg/Th17 balance. DHA alone or in combination with prednisone significantly inhibited Th17 cell differentiation while induced Treg cell differentiation in vitro. DHA alone or in combination with prednisone also reduced the transcription of RORγt and increased Foxp3 in lymphocytes, as well as IL-17 and TGF-ß levels. Our data indicated that DHA can produce synergistic effect with prednisone to attenuate the symptoms of SLE by restoring Treg/Th17 balance.

[Soft tissue regeneration after implantation of two novel citric acid-based biodegradable materials in rats].

OBJECTIVE: To observe the soft tissue regeneration after implantation of two novel citric acid-based biodegradable materials in the skull defects in rats. METHODS: Two novel citric acid-based biodegradable materials were implanted in the muscular tissues in the thigh and harvested 2 weeks later. Another 40 rats with surgically induced cranial defect were randomized into control group, autograft group, CUPE-HA group, and POC-HA group (n=10), and 3 months after implantation, the materials were harvested for histological and morphometric analyses. RESULTS: Soft tissue regeneration was stimulated by the two biodegradable materials in the muscular tissues. The implants also stimulated angiogenesis and soft tissue regeneration in the cranial defect and accelerated of intramembranous ossification. CONCLUSION: The 2 novel citric acid-based biodegradable materials can induce angiogenesis and soft tissue regeneration and accelerate intramembranous ossification in rats with cranial defects.