Journal: Molecular Medicine Reports

Article Title: Penehyclidine hydrochloride alleviates lipopolysaccharide-induced acute respiratory distress syndrome in cells via regulating autophagy-related pathway

doi: 10.3892/mmr.2020.11739

Figure Lengend Snippet: 3-MA reverses the effects of PHC on LPS-induced BEAS-2B cell proliferation and apoptosis. Following treatment with 10 ng/ml LPS, BEAS-2B cells were treated with 0.05 µM PHC, 100 nM Rapa or 10 mM 3-MA. (A) Cell proliferation was examined by performing the Cell Counting Kit-8 assay. (B) Apoptosis was assessed via Hoechst staining. Red arrows indicate apoptotic cells. magnification, ×200. (C) Flow cytometry analysis was conducted to evaluate the effects of PHC, Rapa and 3-MA on LPS-induced BEAS-2B cell apoptosis and the rate of apoptosis was calculated. (D) Bax, Bcl-2 and caspase-3 expression levels were measured by western blotting and semi-quantified. **P<0.01 vs. control group; ## P<0.01 vs. LPS group; && P<0.01 vs. LPS + PHC group. 3-MA, 3-methyladenine; PHC, penehyclidine hydrochloride; LPS, lipopolysaccharide; Rapa, rapamycin.

Article Snippet: Alternatively, following treatment with 0.05 μM PHC, LPS-induced BEAS-2B cells were treated with 100 nM autophagy agonist rapamycin (Rapa; cat. no. IMT1001-100MG; Gene Operation) or 10 mM autophagy inhibitor (3-MA; cat. no. S2767-1; Selleck Chemicals).

Techniques: Cell Counting, Staining, Flow Cytometry, Expressing, Western Blot, Control

Journal: Molecular Medicine Reports

Article Title: Penehyclidine hydrochloride alleviates lipopolysaccharide-induced acute respiratory distress syndrome in cells via regulating autophagy-related pathway

doi: 10.3892/mmr.2020.11739

Figure Lengend Snippet: PHC weakens ARDS damage by regulating the autophagy pathway. LPS-induced BEAS-2B cells were treated with PHC, Rapa or 3-MA. ELISAs were performed to analyze alterations to (A) IFN-γ, (B) TNF-α, (C) IL-1β, (D) IL-6 and (E) NO levels. *P<0.05 and **P<0.01 vs. control group; # P<0.05 and ## P<0.01 vs. LPS group; & P<0.05 and && P<0.01 vs. LPS + PHC group. PHC, penehyclidine hydrochloride; ARDS, acute respiratory distress syndrome; LPS, lipopolysaccharide; Rapa, rapamycin; IFN, interferon; TNF, tumor necrosis factor; IL, interleukin; NO, nitric oxide.

Article Snippet: Alternatively, following treatment with 0.05 μM PHC, LPS-induced BEAS-2B cells were treated with 100 nM autophagy agonist rapamycin (Rapa; cat. no. IMT1001-100MG; Gene Operation) or 10 mM autophagy inhibitor (3-MA; cat. no. S2767-1; Selleck Chemicals).

Techniques: Control

Journal: Molecular Medicine Reports

Article Title: Penehyclidine hydrochloride alleviates lipopolysaccharide-induced acute respiratory distress syndrome in cells via regulating autophagy-related pathway

doi: 10.3892/mmr.2020.11739

Figure Lengend Snippet: 3-MA reverses the effects of PHC on LPS-induced BEAS-2B cell autophagy. BEAS-2B cells were treated with 0.05 µM PHC, 100 nM Rapa or 10 mM 3-MA after treatment with 10 ng/ml LPS. (A) Immunofluorescence assays were conducted to assess LC3B expression. Green indicates LC3B expression and blue (DAPI) indicates the nuclei (magnification, ×200). (B) Autophagosomes were observed using a transmission electron microscope at ×40,000 magnification. LC3B, p62 and Beclin-1 expression levels were (C) determined by western blotting and (D) semi-quantified. **P<0.01 vs. control group; # P<0.05 and ## P<0.01 vs. LPS group; & P<0.05 and && P<0.01 vs. LPS + PHC group. 3-MA, 3-methyladenine; PHC, penehyclidine hydrochloride; LPS, lipopolysaccharide; Rapa, rapamycin; LC3, light chain 3.

Article Snippet: Alternatively, following treatment with 0.05 μM PHC, LPS-induced BEAS-2B cells were treated with 100 nM autophagy agonist rapamycin (Rapa; cat. no. IMT1001-100MG; Gene Operation) or 10 mM autophagy inhibitor (3-MA; cat. no. S2767-1; Selleck Chemicals).

Techniques: Immunofluorescence, Expressing, Transmission Assay, Microscopy, Western Blot, Control