Discovery based on SPR drug chip that corilagin alleviates acute lung injury in mice by inhibiting necroptosis through targeting RIPK1/RIPK3/MLKL pathway.

Fucosterol attenuates lipopolysaccharide-induced acute lung injury in mice

In this study, we found that chloroform fraction (CF) from TJP ethanolic extract inhibited lipopolysaccharide (LPS)-induced production of nitric oxide (NO) and intracellular ROS in RAW264.7 cells. In addition, expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) genes was reduced, as evidenced by western blot. Our results indicate that CF exerts anti-inflammatory effects by down-regulating expression of iNOS and COX-2 genes through inhibition of MAPK (ERK, JNK and p38) and NF-κB signaling. Similarly we also evaluated the effects of CF on LPS-induced acute lung injury. Male Balb/c mice were pretreated with dexamethasone or CF 1 h before intranasal instillation of LPS. Eight hours after LPS administration, the inflammatory cells in the bronchoalveolar lavage fluid (BALF) were determined. The results indicated that CF inhibited LPS-induced TNF-α and IL-6 production in a dose dependent manner. It was also observed that CF attenuated LPS-induced lung histopathologic changes. In conclusion, these data demonstrate that the protective effect of CF on LPS-induced acute lung injury (ALI) in mice might relate to the suppression of excessive inflammatory responses in lung tissue. Thus, it can be suggested that CF might be a potential therapeutic agent for ALI.

In this study, the anti-inflammatory effects of the methanolic extract (TE) of Plumeria obtusa L. (aerial parts) and its fractions were evaluated in vitro, and active fraction was evaluated in vivo. Among tested extracts, dichloromethane fraction (DCM-F) exhibited the strongest inhibition of lipopolysaccharide (LPS)-induced nitric oxide (NO) in RAW 264.7 macrophages. The effect of DCM-F on LPS-induced acute lung injury (ALI) in mice was studied. The animals were divided into five groups (n = 7) randomly; Gp I: negative control, GP II: positive control (LPS group), GP III: standard (dexamethasone, 2 mg/kg b.wt), GP IV and V: DCM-F (100 mg/kg), and DEM-F (200 mg/kg), respectively. DCM-F at a dose of 200 mg/kg suppressed the ability of LPS to increase the levels of nitric oxide synthase (iNOS), NO, tumor necrosis factor-α (TNF-α), and interleukin 6 (IL-6), as measured by ELISA. In addition, the expression of cyclooxygenase-2 (COX-2) was reduced (determined by immunohistochemistry) and the level of malondialdehyde (MDA) was decreased while that of catalase was restored to the normal values. Furthermore, the histopathological scores of inflammation induced by LPS were reduced. Twenty-two compounds were tentatively identified in DCM-F using LC/ESI-QToF with iridoids, phenolic derivatives and flavonoids as major constituents. Identified compounds were subjected to two different molecular docking processes against iNOS and prostaglandin E synthase-1 target receptors. Notably, protoplumericin A and 13-O-coumaroyl plumeride were the most promising members compared to the co-crystallized inhibitor in each case. These findings suggested that DCM-F attenuates the LPS-induced ALI in experimental animals through its anti-inflammatory and antioxidant potential.

The neurotoxicant PCB-95 by increasing the neuronal transcriptional repressor REST down-regulates caspase-8 and increases Ripk1, Ripk3 and MLKL expression determining necroptotic neuronal death

Euphorbia cuneata (EC; Euphorbiaceae), which widely grows in Saudi Arabia and Yemen, is used traditionally to treat pain and inflammation. This study aimed to evaluate the protective anti-inflammatory effect of a standardized extract of EC against lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice and the possible underlying mechanism(s) of this pharmacologic activity. ALI was induced in male Balb/c mice using intraperitoneal injection of LPS. A standardized total methanol extract of EC or dexamethasone was administered 5 days prior to LPS challenge. Bronchoalveolar fluid (BALF) and lung samples were collected for analysis. The results demonstrated the protective anti-inflammatory effect of EC against LPS-induced ALI in mice. Standardized EC contained 2R-naringenin-7-O-β-glucoside (1), kaempferol-7-O-β-glucoside (2), cuneatannin (3), quercetin (4), and 2R-naringenin (5) in concentrations of 6.16, 4.80, 51.05, 13.20, and 50.00 mg/g of extract, respectively. EC showed a protective effect against LPS-induced pulmonary damage. EC reduced lung wet/dry weight (W/D) ratio and total protein content in BALF, indicating attenuation of the pulmonary edema. Total and differential cell counts were decreased in EC-treated animals. Histopathological examination confirmed the protective effect of EC, as indicated by an amelioration of LPS-induced lesions in lung tissue. EC also showed a potent anti-oxidative property as it decreased lipid peroxidation and increased the antioxidants in lung tissue. Finally, the anti-inflammatory activity of EC was obvious through its ability to suppress the activation of nuclear factor-κB (NF-κB), and hence its reduction of the levels of downstream inflammatory mediators. In conclusion, these results demonstrate the protective effects of EC against LPS-induced lung injury in mice, which may be due to its antioxidative and anti-inflammatory activities.

Suppression of NF-κB pathway by crocetin contributes to attenuation of lipopolysaccharide-induced acute lung injury in mice

Protective effects of total alkaloids from Dendrobium crepidatum against LPS-induced acute lung injury in mice and its chemical components

Aim of the study: Receptor-interacting protein (RIP) kinase family members are involved in several biological processes. However, their role in acute lung injury (ALI) is still unclear. In the present study, we aim to determine the expression and function of RIP kinase family in ALI. Materials and methods: In the present study, ALI was induced in BALB/c male mice by intravenously injecting lipopolysaccharide (LPS). The expression levels of the RIP kinase family in ALI mice were determined using western blotting and immunohistochemical staining. The specific RIP-1 inhibitor, necrostatin-1, was used to treat LPS-induced ALI mice, followed by survival time recording, as well as histopathological and immunohistochemical staining of lung tissues, western blotting, myeloperoxidase (MPO) assay and enzyme-linked immunosorbent assay (ELISA) of related cytokines and downstream target expression. Results: We found that RIP-1 expression was upregulated in the lung of ALI mice and inhibition of RIP-1 by necrostatin-1 significantly prolonged the survival time of mice, which was accompanied by less serve lung injury. Furthermore, lower expression of pro-inflammatory cytokines (interleukin [IL]-6, tumor necrosis factor [TNF]-α, IL-8, cyclooxygenase [COX]-2, monocyte chemoattractant protein [MCP]-1, and IL-1β), MPO and nuclear factor (NF)-κB activation were found in bronchoalveolar lavage fluid (BALF) and lung tissues of necrostatin-1-treated ALI mice. Necrostatin-1 also attenuated LPS-induced pro-inflammatory cytokine expression and NF-κB activation in RAW 264.7 cells. Conclusions: In summary, necrostatin-1 protected against LPS-induced ALI in mice by inhibiting inflammation and pulmonary NF-κB activation. Thus, necrostatin-1 could be a novel therapeutic strategy for ALI.

Objective To investigate the protective effect of neurotensins on inflammatory responses in mice with LPS-induced acute lung injury. Methods 100 specific-pathogen free C57BL/6 mice (20-25 g, 6-8 weeks, 50 males and 50 females) were obtained from the Shanghai Laboratory Animal Center (SLAC) (Shanghai, China). All animals were randomly divided into five groups:① Normal control group: mice were treated with 60 μl PBS intratracheally;② LPS-induced acute lung injury group: mice were treated with 60 μg LPS intratracheally;③ 20 mg/kg neurotensins treated ALI group: mice were subjected to LPS-induced ALI and treated with 20 mg/kg Nts via tail vein injection 1 hour after LPS challenge;④ 40 mg/kg neurotensins treated ALI group: mice were subjected to LPS-induced ALI and treated with 40 mg/kg Nts via tail vein injection 1 hour after LPS challenge;⑤ 80 mg/kg neurotensins treated ALI group: mice were subjected to LPS-induced ALI and received 80 mg/kg Nts treatment via tail vein injection. The severity of lung injury, MPO acitvity, neutrophils infilatration, lung edema, and pro-inflammatory cytokines concentration in BALF were assessed 24 hours after ALI. Results Compared with the control group of mice, LPS induction significantly increased the severity of lung injury, including the lung edema, inflammatory cell infiltration, and the production of inflammatory cytokines in BALF (TNF-α, IL-6, IL-1β, and MCP-1). However, Neurotensins treatment obviously attenuated the lung injury caused by LPS induction, including the lung edema, the infiltration of inflammatory cells , and the secretion of inflammatory cytokines (TNF-α, IL-6, IL-1β, and MCP-1). Meanwhile, the protective effect is dosage dependent. Conclusion Neurotensins have a protective effect on LPS-induced acute lung injury in mice, and the protective mechanisms may be related to block the tachykinins mediated inflammatory pathways activation. Key words: Neurotensins; Tachykinins; Inflammatory response; Acute lung injury

Necroptosis is a mechanism by which cells can kill themselves that does not require caspase activity or the presence of the pro-apoptotic Bcl-2 family members Bax or Bak. It has been reported that RIPK3 (receptor interacting protein kinase 3) activates MLKL (mixed lineage kinase domain-like) to cause cell death that requires dynamin-related protein 1 (Drp1), because survival was increased in cells depleted of Drp1 or treated with the Drp1 inhibitor mdivi-1. To analyze necroptosis in a system that does not require addition of tumor necrosis factor (TNF), we used a construct that allows RIPK3 to be induced in cells, and then dimerized via an E. coli gyrase domain fused to its carboxyl-terminus, using the dimeric gyrase binding antibiotic coumermycin. We have previously shown elsewhere that RIPK3 dimerized in this manner not only induces necroptosis but also apoptosis, which can be inhibited by the broad-spectrum caspase inhibitor Q-VD-OPh (QVD). In response to RIPK3 dimerization, wild-type mouse embryonic fibroblasts (MEFs) underwent cell death that was reduced but not completely blocked by QVD. In contrast, death upon dimerization of RIPK3 in Mlkl−/− MEFs was completely inhibited with QVD, confirming that MLKL is required for necroptosis. Similar to wild-type MEFs, most Drp1−/− MEFs died when RIPK3 was activated, even in the presence of QVD. Furthermore, overexpression of wild-type MLKL or dominant active mutants of MLKL (Q343A or S345E/S347E) caused death of wild-type and Drp1−/− MEFs that was not inhibited with QVD. These results indicate that necroptosis caused by RIPK3 requires MLKL but not Drp1.

Plantamajoside ameliorates lipopolysaccharide-induced acute lung injury via suppressing NF-κB and MAPK activation

Small extracellular vesicles (sEVs) have been recognized to be more effective than direct stem cell differentiation into functional target cells in preventing tissue injury and promoting tissue repair. Our previous study demonstrated the protective effect of adipose-derived stem cells (ADSCs) on lipopolysaccharide (LPS)-induced acute lung injury and the effect of autophagy on ADSC functions, but the role of ADSC-derived sEVs (ADSC-sEVs) and autophagy-mediated regulation of ADSC-sEVs in LPS-induced pulmonary microvascular barrier damage remains unclear. After treatment with sEVs from ADSCs with or without autophagy inhibition, LPS-induced human pulmonary microvascular endothelial cell (HPMVECs) barrier damage was detected. LPS-induced acute lung injury in mice was assessed in vivo after intravenous administration of sEVs from ADSCs with or without autophagy inhibition. The effects of autophagy on the bioactive miRNA components of ADSC-sEVs were assessed after prior inhibition of cell autophagy. We found that ADSC-sEV effectively alleviated LPS-induced apoptosis, tight junction damage and high permeability of PMVECs. Moreover, in vivo administration of ADSC-sEV markedly inhibited LPS-triggered lung injury. However, autophagy inhibition, markedly weakened the therapeutic effect of ADSC-sEVs on LPS-induced PMVECs barrier damage and acute lung injury. In addition, autophagy inhibition, prohibited the expression of five specific miRNAs in ADSC-sEVs -under LPS-induced inflammatory conditions. Our results indicate that ADSC-sEVs protect against LPS-induced pulmonary microvascular barrier damage and acute lung injury. Autophagy is a positive mediator of sEVs function, at least in part through controlling the expression of bioactive miRNAs in sEVs.

The protective effect of hyperin on LPS-induced acute lung injury in mice

The aim of this study was to evaluate the effects of single oral doses of D-005 (a lipid extract obtained from the fruit oil of Acrocomia crispa) on LPS-induced acute lung injury (ALI) in mice. D-005 batch composition was: lauric (35.8%), oleic (28.4%), myristic (14.2%), palmitic (8.9%), stearic (3.3%), capric (1.9%), caprylic (1.2%), and palmitoleic (0.05%) acids, for a total content of fatty acids of 93.7%. D-005 (200 mg/kg) significantly reduced lung edema (LE) (≈ 28% inhibition) and Lung Weight/Body Weight ratio (LW/BW) (75.8% inhibition). D-005 (25, 50, 100 and 200 mg/kg) produced a significant reduction of Histological score (59.9, 56.1, 53.5 and 73.3% inhibition, respectively). Dexamethasone, as the reference drug, was effective in this experimental model. In conclusion, pretreatment with single oral doses of D-005 significantly prevented the LPS-induced ALI in mice.

Monoammonium glycyrrhizinate inhibited the inflammation of LPS-induced acute lung injury in mice