Chalcone derivatives ameliorate lipopolysaccharide-induced acute lung injury and inflammation by targeting MD2.

Acute lung injury (ALI) and its severe form acute respiratory distress syndrome (ARDS) are known as the common causes of respiratory failure in critically ill patients. Myeloid differentiation 2 (MD2), a co-receptor of toll like receptor 4 (TLR4), plays an important role in LPS-induced ALI in mice. Since MD2 inhibition by pharmacological inhibitors or gene knockout significantly attenuates ALI in animal models, MD2 has become an attractive target for the treatment of ALI. In this study we identified two chalcone-derived compounds, 7w and 7x, as new MD2 inhibitors, and investigated the therapeutic effects of 7x and 7w in LPS-induced ALI mouse model. In molecular docking analysis we found that 7w and 7x, formed pi-pi stacking interactions with Phe151 residue of the MD2 protein. The direct binding was confirmed by surface plasmon resonance analysis (with KD value of 96.2 and 31.2 µM, respectively) and by bis-ANS displacement assay. 7w and 7x (2.5, 10 µM) also dose-dependently inhibited the interaction between lipopolysaccharide (LPS) and rhMD2 and LPS-MD2-TLR4 complex formation. In mouse peritoneal macrophages, 7w and 7x (1.25-10 µM) dose-dependently inhibited LPS-induced inflammatory responses, MAPKs (JNK, ERK and P38) phosphorylation as well as NF-κB activation. Finally, oral administration of 7w or 7x (10 mg ·kg-1 per day, for 7 days prior LPS challenge) in ALI mouse model significantly alleviated LPS-induced lung injury, pulmonary edema, lung permeability, inflammatory cells infiltration, inflammatory cytokines expression and MD2/TLR4 complex formation. In summary, we identify 7w and 7x as new MD2 inhibitors to inhibit inflammatory response both in vitro and in vivo, proving the therapeutic potential of 7w and 7x for ALI and inflammatory diseases.

TLR2 regulates angiotensin II-induced vascular remodeling and EndMT through NF-κB signaling.

Excessive vascular remodeling has been shown in hypertensive patients. In experimental models of hypertensive vascular injury, such as angiotensin II (Ang II) challenged mice, toll like receptor 2 (TLR2) initiates inflammatory responses. More recently, studies have reported atypical endothelial to mesenchymal transition (EndMT) in vascular injuries and inflammatory conditions. Here, we aimed to investigate whether TLR2 mediates Ang II-induced vascular inflammation and initiates EndMT. In a mouse model of angiotensin II-induced hypertension, we show that aortas exhibit increased medial thickening, fibrosis, and features of EndMT. These alterations were not observed in TLR2 knockout mice in response to Ang II. TLR2 silencing in cultured endothelial cells confirmed the essential role of TLR2 in Ang II-induced inflammatory factor induction, and EndMT-associated phenotypic change. Mechanistically, we found Ang II activates nuclear factor-κB signaling, inducing pro-inflammatory cytokine production, and mediates EndMT in both cultured endothelial cells and in mice. These studies illustrate a novel role of TLR2 in regulating Ang II-induced deleterious vascular remodeling through the induction of EndMT. The studies also suggest that TLR2 may be targeted to alleviate hypertension-associated vascular injury.

Protective effects of ferroptosis inhibition on high fat diet-induced liver and renal injury in mice.

As a complex and highly prevalent global public health problem, obesity is associated with multiple diseases, including liver and renal injury. As an iron-dependent form of cell death, ferroptosis is different from apoptosis and necrosis, which has been reported to participate in pathologic processes of many diseases. However, whether ferroptosis is involved in obesity-induced liver and renal injury remains unclear. Male C57BL/6 mice were fed with high-fat diet (HFD) or control diet for 16 weeks and treated with 5 mg/kg or 10 mg/kg ferroptosis inhibitor, ferrostatin-1 (Fer-1), for the last 8 weeks with results indicating that glutathione peroxidase 4 (GPX4) gene expression decreased in the liver and renal tissue of obese mice. Additionally, Fer-1 pretreatment prevented the obesity-induced decline of GPX4. More importantly, Fer-1 treatment attenuated HFD-induced pathological and functional impairment, fibrosis, inflammatory cell infiltration, and inflammatory cytokine expression in liver and renal tissues. In short, our results indicate that obesity can induce ferroptosis and ferroptosis inhibitor, Fer-1, thereby inhibiting obesity-induced liver and renal injury in mice. The study provides a new therapeutic direction for the complications of obesity.

Inhibition of MyD88 by LM8 Attenuates Obesity-Induced Cardiac Injury.

Obesity-induced cardiomyopathy involves chronic and sustained inflammation. The toll-like receptor 4 (TLR4) signaling pathway can associate innate immunity with obesity. Myeloid differentiation primary response 88 (MyD88), an indispensable downstream adaptor molecule of TLR4, has been reported to mediate obesity complications. However, whether inhibition of MyD88 can mitigate obesity-induced heart injury remains unclear. LM8, a new MyD88 inhibitor, exhibits prominent anti-inflammatory activity in lipopolysaccharide-treated macrophages. In this study, the protective effects of LM8 on a high-fat diet (HFD)-induced heart injury were assessed in a mouse model of obesity. As suggested from the achieved results, LM8 treatment alleviated HFD-induced pathological and functional damages of the heart in mice. Meantime, the treatment of mice with LM8 could significantly inhibit myocardial hypertrophy, fibrosis, inflammatory cytokines expression, and inflammatory cell infiltration induced by HFD. Besides, LM8 administration inhibited the formation of MyD88/TLR4 complex, phosphorylation of ERK, and activation of nuclear factor-κB induced by HFD. According to the achieved results, MyD88 inhibitor LM8 ameliorated obesity-induced heart injury by inhibiting MyD88-ERK/nuclear factor-κB dependent cardiac inflammatory pathways. Furthermore, targeting MyD88 might be a candidate of a therapeutic method to treat obesity-induced heart injury.