Modulation of macrophages by a phillyrin-loaded thermosensitive hydrogel promotes skin wound healing in mice.

BACKGROUND: Impaired wound healing in traumatic skin injuries remains a severe clinical challenge due to impaired re-vascularization, harmful bacteria infection, and inflammation dysregulation. Macrophages are recognized as prominent immune cells in tissue regeneration and wound healing. Consequently, the modulation of macrophages provides a promising therapeutic target for wound healing disorders. Here, we aimed to explore whether a novel constructed combination of thermosensitive hydrogel Pluronic F-127 (PF-127) and phillyrin (PH, the main active compound of forsythia suspensa) could improve skin wound healing. METHODS: Firstly, the biological effects of pH on the phenotype and inflammation of macrophages were assessed by flow cytometry and ELISA. The biocompatibility of the PF-127 plus PH combination was investigated on keratinocytes and red blood cells. The biological effect of PF-127/PH hydrogel on the migratory ability of keratinocytes in vitro was evaluated using the scratch and transwell migration assays. In addition,S. aureusandE. coliwere employed to test the antibacterial properties of the PF-127 plus PH combination. Finally, PF-127 plus PH scaffold was appliedto the full-thickness skin defect in mice. Histomorphological evaluation and immunochemistry were performed to explore the wound-healing activity of PF-127/PH hydrogel. RESULTS: PH can promote the polarization of macrophages from the M1 (pro-inflammatory) phenotype to the M2 (anti-inflammatory) phenotype. The PF-127/PH hydrogel was highly biocompatible and showed a potent stimulative effect on the migration of keratinocytesin vitro. The combination of PF-127 and PH exerted a pronounced antibacterial activity onS. aureusandE. coli in vitro.PF-127/PH hydrogel potently accelerates the healing of full-thickness skin defects by promoting skin cell proliferation, accelerating angiogenesis, and inhibiting inflammation. CONCLUSIONS: Our study suggests that PF-127/PH hydrogel has excellent potential for treating traumatic skin defects.

Phillyrin sensitizes lung cancer cells to ferroptosis through inhibiting FTH1/SLC7A11 axis.

Ferroptosis is a newly found form of non-apoptotic regulated cell death that is essential for cancer progression. Phillyrin (PHN), an effective lignan glycoside of Forsythia suspensa, has been explored as a potential anticancer agent in some cancer types. However, it is unclear whether and how PHN could promote the death of non-small cell lung cancer (NSCLC) cells by inducing ferroptosis. Our study showed for the first time that PHN induced cell death and attenuated cell proliferation in NSCLC cells in vitro and in vivo. Functional assays showed that ferroptosis was the predominant form that contributed to PHN-induced cell death of NSCLC cells. Mechanistically, NSCLC cells exposed to PHN had a decreased ferritin heavy chain 1 (FTH1) and SLC7A11 protein expression. Exogenetic overexpression of FTH1 substantially abrogated the tumor-inhibiting effects of PHN and further upregulated the expression of SLC7A11 in NSCLC cells. In conclusion, our findings suggest that the natural product PHN exerted its antitumor activity in NSCLC by promoting ferroptosis, and the FTH1/SLC7A11 axis plays an indispensable role in PHN-induced ferroptotic cell death.

Syringaresinol inhibits cardiorenal fibrosis through HSP90 in a cardiorenal syndrome type 2.

BACKGROUND: Syringaresinol processes anti-inflammatory and antioxidative activity. However, the effects of syringaresinol on cardiorenal fibrosis caused by cardiorenal syndrome type 2 (CRS2) are unclear. METHODS: Molecular docking predicted binding activity of syringaresinol to heat shock protein 90 (HSP90). The toxicity of a 4-weeks treatment with 20 mg/kg of syringaresinol was observed by measuring serum pro-inflammatory cytokines levels and by cardiorenal pathology. A CRS2 rad model was established by myocardial infarction using ligation over an 8 week-period. Rats were divided into five groups, including sham, CRS2, pimitespib, syringaresinol, and HSP90 + syringaresinol. Rats were received a 4-weeks daily treatment with 10 mg/kg pimitespib (a HSP90 inhibitor) or 20 mg/kg syringaresinol. Recombinant adeno-associated virus (rAAV) carrying a periostin (PE) promoter driving the expression of wild-type HSP90 (rAAV9-PE-HSP90, 1 × 1011 µg) was treated intravenously once in CRS2 model rats. Cardiorenal function and pathology were assessed. Expressions of HSP90 and TGF-ß1 in the myocardium and kidney were measured by immunohistochemistry and western blotting. RESULTS: Syringaresinol showed good binding activity with HSP90, and no signs of toxicity in rats following treatment. Pimitespib or syringaresinol significantly improved the cardiorenal function and fibrosis in rats with CRS2. Meanwhile, the rAAV9-PE-HSP90 injection obviously blocked the effects of syringaresinol. CONCLUSIONS: Syringaresinol targets HSP90 to suppress CRS2-induced cardiorenal fibrosis, providing a promising therapeutic drug for CRS2.

miR­146a­5p protects against renal injury in MRL/lpr mice via improvement of the Treg/Th17 imbalance by targeting the TRAF6/NF­κB axis.

Dysregulated microRNA (miRNA or miR) expression is an important cause of immune homeostasis disorder in patients with systemic lupus erythematosus and lupus nephritis (LN). The present study evaluated the possibility of using miR-146a-5p as a therapeutic target for treating LN. The effects of miR-146a-5p on lupus syndrome in MRL/lpr mice were evaluated. MRL/lpr mice were injected with miR-146a-5p agomir (M146AG) or agomir negative control (NC). Renal function index, pathology and protein expression levels of inflammatory factors in MRL/lpr mice were evaluated after M146AG or agomir NC treatment. Reverse transcription-quantitative PCR, western blotting and immunofluorescence were used to assess the effect of M146AG on mRNA and protein expression levels of (tumor necrosis factor receptor-associated factor 6) TRAF6/NF-κB axis components. A luciferase dual reporter system was used to assess the mechanism of regulation of TRAF6/NF-κB axis expression. Finally, flow cytometry was used to assess the regulatory effect of M146AG on regulatory T cell (Treg)/T helper 17 (Th17) balance. The findings demonstrated that M146AG ameliorated renal lesions and the inflammatory response in MRL/lpr mice. TRAF6 was demonstrated to be targeted and significantly negatively regulated by miR-146a-5p. M146AG intervention significantly increased expression of miR-146a-5p and significantly downregulated the mRNA and protein expression levels of TRAF6 and NF-κB in CD4+ T cells of MRL/lpr mice. Furthermore, M146AG intervention alleviated Treg/Th17 imbalance in MRL/lpr mice peripheral blood. The present findings demonstrated that M146AG improved Treg/Th17 imbalance and alleviated renal lesions in MRL/lpr mice by targeting the TRAF6/NF-κB axis. This may provide a new theoretical basis for the clinical diagnosis and treatment of LN.