Journal: Respiratory Research

Article Title: Lipoxin A4 receptor agonist BML-111 induces autophagy in alveolar macrophages and protects from acute lung injury by activating MAPK signaling

doi: 10.1186/s12931-018-0937-2

Figure Lengend Snippet: BML-111 alleviated ALI in vivo. ALI model was established in rats by intratracheal instillation of LPS and rats were either not treated (ALI), or treated with vehicle (PBS + ALI) or BML-111 (BML-111 + ALI). As controls, rats not through ALI induction and treated with either vehicle (PBS) or BML-111 were used. a Upon sacrifice, the lung tissue from each group was examined by HE staining and assessed for ALI score. b The lung tissue was measured for wet/dry weight ratio and compared among different groups. n = 6, * P

Article Snippet: Taken together, the data suggest that BML-111 is sufficient to promote autophagy of AMs and thus may protect these cells from LPS-induced apoptosis.

Techniques: In Vivo, Staining

Journal: Respiratory Research

Article Title: Lipoxin A4 receptor agonist BML-111 induces autophagy in alveolar macrophages and protects from acute lung injury by activating MAPK signaling

doi: 10.1186/s12931-018-0937-2

Figure Lengend Snippet: The benefits of BML-111 were associated with reduced inflammation and enhanced autophagy in vivo. Bronchoalveolar lavage was collected from rats of each group and the levels of TNF-α ( a ) and IL-6 ( b ) was measured using ELISA. AMs were isolated from rats of each group. c The expressions of TNF-α and IL-6 on the steady-state mRNA level were measured by RT-qPCR. d The expressions of BECN1, SQSTM1/p62, LC3-I, and LC3-II in isolated AM were examined by Western blot. Representative Western blot image was shown on the left and the quantification of each protein level relative to that of the internal control (GAPDH) shown on the right ( e ). n = 6, * P

Article Snippet: Taken together, the data suggest that BML-111 is sufficient to promote autophagy of AMs and thus may protect these cells from LPS-induced apoptosis.

Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Affinity Magnetic Separation, Isolation, Quantitative RT-PCR, Western Blot

Journal: Respiratory Research

Article Title: Lipoxin A4 receptor agonist BML-111 induces autophagy in alveolar macrophages and protects from acute lung injury by activating MAPK signaling

doi: 10.1186/s12931-018-0937-2

Figure Lengend Snippet: BML-111 inhibited LPS-induced apoptosis. AM were isolated from rats and treated with either vehicle (PBS), LPS (to induce ALI), BML-111, BML-111 + LPS. a At 24 h after the treatment, the cell viability was examined by MTT assay. b The apoptosis of cells was determined by flow cytometry following staining the cells with Annexin V and PI. c The expression of different apoptosis biomarkers, including cleaved-Caspase 3, cleaved-Caspase 8,cleaved-Caspase 9, cleaved-PARP, Bcl-2, and Bax was detected by Western blot. Representative Western blot image was shown on the left and the quantification of each protein level relative to that of the internal control (GAPDH) shown on the right. n = 3, * P

Article Snippet: Taken together, the data suggest that BML-111 is sufficient to promote autophagy of AMs and thus may protect these cells from LPS-induced apoptosis.

Techniques: Isolation, MTT Assay, Flow Cytometry, Cytometry, Staining, Expressing, Western Blot

Journal: Respiratory Research

Article Title: Lipoxin A4 receptor agonist BML-111 induces autophagy in alveolar macrophages and protects from acute lung injury by activating MAPK signaling

doi: 10.1186/s12931-018-0937-2

Figure Lengend Snippet: BML-111 elevated LC3-II level in AMs. AMs were treated with increasing concentrations of BML-111 for 2 h ( a ) or with 100 nM of BML-111 for indicated time periods ( b ). The expression of LC3-I and LC3-II was examined by Western blot. Representative Western blot image was shown on the top and the LC3-II/LC3-I ratio shown on the bottom. n = 3, * P

Article Snippet: Taken together, the data suggest that BML-111 is sufficient to promote autophagy of AMs and thus may protect these cells from LPS-induced apoptosis.

Techniques: Affinity Magnetic Separation, Expressing, Western Blot

Journal: Respiratory Research

Article Title: Lipoxin A4 receptor agonist BML-111 induces autophagy in alveolar macrophages and protects from acute lung injury by activating MAPK signaling

doi: 10.1186/s12931-018-0937-2

Figure Lengend Snippet: BML-111 targeted MAPK1 pathway but mTOR-independent mechanism to induce autophagy. a The activation of MAPK1 and MAPK8 was detected by Western blot in AM treated as indicated. Representative Western blot image was shown on the left and the quantification of each protein level relative to that of the internal control (GAPDH) shown on the right. b AM were treated as indicated, in autophagy inhibitor MHY-1485 and mTOR inhibitor Rapamycin. LC3-II expression was examined by immunofluorescence (green signals). All cells were counter stained with DAPI (blue signal). Representative immunofluorescence images from indicated cells were shown on the bottom and the percentage of LC3-II+ cells quantified and shown as histogram on the top. n = 3, * P

Article Snippet: Taken together, the data suggest that BML-111 is sufficient to promote autophagy of AMs and thus may protect these cells from LPS-induced apoptosis.

Techniques: Activation Assay, Western Blot, Expressing, Immunofluorescence, Staining

Journal: Respiratory Research

Article Title: Lipoxin A4 receptor agonist BML-111 induces autophagy in alveolar macrophages and protects from acute lung injury by activating MAPK signaling

doi: 10.1186/s12931-018-0937-2

Figure Lengend Snippet: BML-111 elevated autophagy level in LPS-treated AM. AMs were treated as indicated. a The expression of LC3-I and LC3-II was examined by Western blot. Representative Western blot image was shown on the top and the LC3-II/LC3-I ratio shown on the bottom. b The expression of LC3-II in AM was detected by immunofluorescence (green signal). All cells were counter stained with DAPI (blue signal). Representative immunofluorescence images from indicated cells were shown on the left and the percentage of LC3-II+ cells quantified and shown as histogram on the right. c The expression of different autophagy and apoptosis biomarkers, including BECN1, SQSTM1/p62 was detected by Western blot. n = 3, * P

Article Snippet: Taken together, the data suggest that BML-111 is sufficient to promote autophagy of AMs and thus may protect these cells from LPS-induced apoptosis.

Techniques: Affinity Magnetic Separation, Expressing, Western Blot, Immunofluorescence, Staining

Journal: Toxicology Research

Article Title: Development of a BALB/c mouse model for food allergy: comparison of allergy-related responses to peanut agglutinin, β-lactoglobulin and potato acid phosphatase

doi: 10.1039/c6tx00371k

Figure Lengend Snippet: The histopathology results of jejunum segments in mice (sensitized with 0.05 mg and 0 mg protein) i.p. and i.g. challenged with OVA, PNA, β-LG and PAP, respectively. Severe jejunum lesions are linked to clinical signs of systemic anaphylaxis in this model. The solid arrow in (a) indicates inflammatory cell infiltration in submucosa; the solid arrow in (c) indicates villi swelling; the solid arrows in (e) and (i) indicate structure looseness. (a), (e), (i), (m) Ear segments from mice i.p. sensitized and i.p. challenged with OVA, PNA, β-LG and PAP, respectively. (b), (f), (j), (n) Ear segments from control mice challenged with OVA, PNA, β-LG and PAP, respectively. (c), (g), (k), (o) Ear segments from mice i.p. sensitized and i.g. challenged with OVA, PNA, β-LG and PAP, respectively. (d), (h), (l), (p) Ear segments from control mice i.g. challenged with OVA, PNA, β-LG and PAP, respectively.

Article Snippet: Ovalbumin (OVA, catalogue number: A5503), peanut agglutinin (PNA, catalogue number: L0881), β-lactoglobulin (β-LG, catalogue number: L3908) and potato acid phosphatase (PAP, catalogue number: P3752) were obtained from Sigma-Aldrich Co. LLC. (St Louis, USA), and reconstituted in phosphate buffered saline (PBS) at 0.2 mg mL–1 (10 mg protein was solubilized in 50 mL PBS), 4 mg mL–1 (40 mg protein was solubilized in 10 mL PBS) and 200 mg mL–1 (1000 mg protein was solubilized in 5 mL PBS), respectively.

Techniques: Histopathology, Mouse Assay

Journal: Toxicology Research

Article Title: Development of a BALB/c mouse model for food allergy: comparison of allergy-related responses to peanut agglutinin, β-lactoglobulin and potato acid phosphatase

doi: 10.1039/c6tx00371k

Figure Lengend Snippet: The histopathology results of lung segments in mice (sensitized with 0.05 mg and 0 protein) i.p. and i.g. challenged with OVA, PNA, β-LG and PAP, respectively. Severe lung lesions are bound up with clinical signs of systemic anaphylaxis in this model. Solid arrows indicate inflammatory cell infiltration. (a), (e), (i), (m) Ear segments from mice i.p. sensitized and i.p. challenged with OVA, PNA, β-LG and PAP, respectively. (b), (f), (j), (n) Ear segments from control mice challenged with OVA, PNA, β-LG and PAP, respectively. (c), (g), (k), (o) Ear from i.p. sensitized and i.g. challenged with OVA, PNA, β-LG and PAP, respectively. (d), (h), (l), (p) Ear segments from control mice i.g. challenged with OVA, PNA, β-LG and PAP, respectively.

Article Snippet: Ovalbumin (OVA, catalogue number: A5503), peanut agglutinin (PNA, catalogue number: L0881), β-lactoglobulin (β-LG, catalogue number: L3908) and potato acid phosphatase (PAP, catalogue number: P3752) were obtained from Sigma-Aldrich Co. LLC. (St Louis, USA), and reconstituted in phosphate buffered saline (PBS) at 0.2 mg mL–1 (10 mg protein was solubilized in 50 mL PBS), 4 mg mL–1 (40 mg protein was solubilized in 10 mL PBS) and 200 mg mL–1 (1000 mg protein was solubilized in 5 mL PBS), respectively.

Techniques: Histopathology, Mouse Assay

Journal: Toxicology Research

Article Title: Development of a BALB/c mouse model for food allergy: comparison of allergy-related responses to peanut agglutinin, β-lactoglobulin and potato acid phosphatase

doi: 10.1039/c6tx00371k

Figure Lengend Snippet: The histopathology results of mice ear segments (sensitized with 0.05 mg and 0 mg protein) i.p. and i.g. challenged with OVA, PNA, β-LG and PAP, respectively. Severe ear damage is linked to clinical signs of systemic anaphylaxis in this model. Solid arrows indicate edema. (a), (e), (i), (m) Ear segments from mice i.p. sensitized and i.p. challenged with OVA, PNA, β-LG and PAP, respectively. (b), (f), (j), (n) Ear segments from control mice challenged with OVA, PNA, β-LG and PAP, respectively. (c), (g), (k), (o) Ear segments from i.p. sensitized and i.g. challenged with OVA, PNA, β-LG and PAP, respectively. (d), (h), (l), (p) Ear segments from control mice i.g. challenged with OVA, PNA, β-LG and PAP, respectively.

Article Snippet: Ovalbumin (OVA, catalogue number: A5503), peanut agglutinin (PNA, catalogue number: L0881), β-lactoglobulin (β-LG, catalogue number: L3908) and potato acid phosphatase (PAP, catalogue number: P3752) were obtained from Sigma-Aldrich Co. LLC. (St Louis, USA), and reconstituted in phosphate buffered saline (PBS) at 0.2 mg mL–1 (10 mg protein was solubilized in 50 mL PBS), 4 mg mL–1 (40 mg protein was solubilized in 10 mL PBS) and 200 mg mL–1 (1000 mg protein was solubilized in 5 mL PBS), respectively.

Techniques: Histopathology, Mouse Assay

Journal: Toxicology Research

Article Title: Development of a BALB/c mouse model for food allergy: comparison of allergy-related responses to peanut agglutinin, β-lactoglobulin and potato acid phosphatase

doi: 10.1039/c6tx00371k

Figure Lengend Snippet: Plasma histamine levels in the protein and control groups. Female BALB/c mice were treated five times intraperitoneally with 0.05 mg and 0 mg protein in 0.25 mL PBS. On day 30, two methods of elicitation of allergic reaction were compared: intraperitoneal injection and intragastric administration. The mice were challenged by intraperitoneal injection (10 mice in the protein group and 10 control mice) with 1 mg protein (OVA, PNA, β-LG and PAP, respectively) in 0.25 mL PBS (that is, 4 mg mL –1 of protein solution), and were challenged by intragastric administration (10 mice in the protein group and 10 control mice) of 50 mg of protein (OVA, PNA, β-LG and PAP, respectively) in 0.25 mL PBS (that is, 0.25 mL of 200 mg mL –1 protein solution). ** represents P

Article Snippet: Ovalbumin (OVA, catalogue number: A5503), peanut agglutinin (PNA, catalogue number: L0881), β-lactoglobulin (β-LG, catalogue number: L3908) and potato acid phosphatase (PAP, catalogue number: P3752) were obtained from Sigma-Aldrich Co. LLC. (St Louis, USA), and reconstituted in phosphate buffered saline (PBS) at 0.2 mg mL–1 (10 mg protein was solubilized in 50 mL PBS), 4 mg mL–1 (40 mg protein was solubilized in 10 mL PBS) and 200 mg mL–1 (1000 mg protein was solubilized in 5 mL PBS), respectively.

Techniques: Mouse Assay, Injection

Journal: Toxicology Research

Article Title: Development of a BALB/c mouse model for food allergy: comparison of allergy-related responses to peanut agglutinin, β-lactoglobulin and potato acid phosphatase

doi: 10.1039/c6tx00371k

Figure Lengend Snippet: Schematic of the sensitization and challenge in this study. Groups of mice ( n = 30) were intraperitoneally sensitized with 0.25 mL of 0.2 mg mL –1 of protein (OVA, PNA, β-LG and PAP, respectively) solutions on days 0, 3, 6, 9, 12. On day 28, ten mice were randomly chosen from each group, then their blood samples were collected, spleen cells were cultured and cytokines were analyzed. On day 30, the challenge was performed and two methods of elicitation of allergic reaction (10 mice for intraperitoneal injection and 10 mice for intragastric administration in each protein group) were compared through the following parameters: symptoms of hypersensitivity, plasma histamine level and histopathologic analysis. The mice in the control group were intraperitoneally sensitized with PBS and challenged with the corresponding proteins in two methods of elicitation.

Article Snippet: Ovalbumin (OVA, catalogue number: A5503), peanut agglutinin (PNA, catalogue number: L0881), β-lactoglobulin (β-LG, catalogue number: L3908) and potato acid phosphatase (PAP, catalogue number: P3752) were obtained from Sigma-Aldrich Co. LLC. (St Louis, USA), and reconstituted in phosphate buffered saline (PBS) at 0.2 mg mL–1 (10 mg protein was solubilized in 50 mL PBS), 4 mg mL–1 (40 mg protein was solubilized in 10 mL PBS) and 200 mg mL–1 (1000 mg protein was solubilized in 5 mL PBS), respectively.

Techniques: Mouse Assay, Cell Culture, Injection