Anti-inflammatory effects of Torin2 on lipopolysaccharide-treated RAW264.7 murine macrophages and potential mechanisms
Context: Torin2 has various pharmacological properties. However, its anti-inflammatory activity has not been reported. Objective: This study focused on the potential anti-inflammatory properties of Torin2 in lipopolysaccharide (LPS)-evoked RAW264.7 murine macrophages. The study aimed to shed light o...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-07-01
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| Series: | Heliyon |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405844022012051 |
| _version_ | 1818017634442018816 |
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| author | Shanshan Wei Ting Wang Qi Cao Xue Chen Zhiyong Chu Zhenzhen Zhang |
| author_facet | Shanshan Wei Ting Wang Qi Cao Xue Chen Zhiyong Chu Zhenzhen Zhang |
| author_sort | Shanshan Wei |
| collection | DOAJ |
| description | Context: Torin2 has various pharmacological properties. However, its anti-inflammatory activity has not been reported. Objective: This study focused on the potential anti-inflammatory properties of Torin2 in lipopolysaccharide (LPS)-evoked RAW264.7 murine macrophages. The study aimed to shed light on the molecular mechanisms that ameliorate these effects. Methods: Torin2 was applied to 100 ng/mL lipopolysaccharide-induced RAW 264.7 macrophages in vitro. Nitric oxide (NO) levels were detected using the Griess reagent kit. Prostaglandin E2 (PGE2), pro-inflammatory cytokines interleukin (IL)-1β, interleukin (IL)-6, and tumor necrosis factor in the supernatant fraction were determined using enzyme-linked immunosorbent assay (ELISA). Gene expression of pro-inflammatory cytokines, cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS) were tested using real-time quantitative polymerase chain reaction (qPCR). Cyclooxygenase-2 and inducible nitric oxide synthase proteins, phosphorylation of mitogen-activated protein kinase (MAPK) subgroups, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), p38, I-kappa-B-alpha (IκBα), and nuclear factor-kappa-B (NF-κB), and activation in extracts were detected via western blotting. Nuclear factor-kappa-B/p65 nuclear translocation was tested using an immunofluorescence assay. Results: The results demonstrated that pre-treatment with Torin2 profoundly attenuated the lipopolysaccharide-stimulated levels of nitric oxide and prostaglandin E2, pro-inflammatory cytokines, messenger ribonucleic acid (mRNA), and protein expression of cyclooxygenase-2 and inducible nitric oxide synthase. Collectively, Torin2 pre-treatment notably weakened lipopolysaccharide-induced damage by reducing the phosphorylation of nuclear factor-kappa-B, p38, c-Jun N-terminal kinase, extracellular signal-regulated kinase proteins, and nuclear factor-kappa-B/p65 nuclear translocation. Conclusion: Numerous pieces of evidence indicated that Torin2 reversed inflammatory activation by regulating nuclear factor-kappa-B and mitogen-activated protein kinase signaling pathways and provided a tentative potential candidate for preventing and treating inflammatory diseases. |
| first_indexed | 2024-04-14T07:29:28Z |
| format | Article |
| id | doaj.art-bbc8bd55d7e947d5a300cda38ea2cf03 |
| institution | Directory Open Access Journal |
| issn | 2405-8440 |
| language | English |
| last_indexed | 2024-04-14T07:29:28Z |
| publishDate | 2022-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Heliyon |
| spelling | doaj.art-bbc8bd55d7e947d5a300cda38ea2cf032022-12-22T02:05:54ZengElsevierHeliyon2405-84402022-07-0187e09917Anti-inflammatory effects of Torin2 on lipopolysaccharide-treated RAW264.7 murine macrophages and potential mechanismsShanshan Wei0Ting Wang1Qi Cao2Xue Chen3Zhiyong Chu4Zhenzhen Zhang5Naval Medical Research Institute, Naval Medical University, Shanghai, 200433, ChinaNaval Medical Research Institute, Naval Medical University, Shanghai, 200433, ChinaSchool of Pharmacy, Naval Medical University, Shanghai, 200433, ChinaSchool of Pharmacy, Naval Medical University, Shanghai, 200433, ChinaNaval Medical Research Institute, Naval Medical University, Shanghai, 200433, China; Corresponding author.Naval Medical Research Institute, Naval Medical University, Shanghai, 200433, China; Corresponding author.Context: Torin2 has various pharmacological properties. However, its anti-inflammatory activity has not been reported. Objective: This study focused on the potential anti-inflammatory properties of Torin2 in lipopolysaccharide (LPS)-evoked RAW264.7 murine macrophages. The study aimed to shed light on the molecular mechanisms that ameliorate these effects. Methods: Torin2 was applied to 100 ng/mL lipopolysaccharide-induced RAW 264.7 macrophages in vitro. Nitric oxide (NO) levels were detected using the Griess reagent kit. Prostaglandin E2 (PGE2), pro-inflammatory cytokines interleukin (IL)-1β, interleukin (IL)-6, and tumor necrosis factor in the supernatant fraction were determined using enzyme-linked immunosorbent assay (ELISA). Gene expression of pro-inflammatory cytokines, cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS) were tested using real-time quantitative polymerase chain reaction (qPCR). Cyclooxygenase-2 and inducible nitric oxide synthase proteins, phosphorylation of mitogen-activated protein kinase (MAPK) subgroups, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), p38, I-kappa-B-alpha (IκBα), and nuclear factor-kappa-B (NF-κB), and activation in extracts were detected via western blotting. Nuclear factor-kappa-B/p65 nuclear translocation was tested using an immunofluorescence assay. Results: The results demonstrated that pre-treatment with Torin2 profoundly attenuated the lipopolysaccharide-stimulated levels of nitric oxide and prostaglandin E2, pro-inflammatory cytokines, messenger ribonucleic acid (mRNA), and protein expression of cyclooxygenase-2 and inducible nitric oxide synthase. Collectively, Torin2 pre-treatment notably weakened lipopolysaccharide-induced damage by reducing the phosphorylation of nuclear factor-kappa-B, p38, c-Jun N-terminal kinase, extracellular signal-regulated kinase proteins, and nuclear factor-kappa-B/p65 nuclear translocation. Conclusion: Numerous pieces of evidence indicated that Torin2 reversed inflammatory activation by regulating nuclear factor-kappa-B and mitogen-activated protein kinase signaling pathways and provided a tentative potential candidate for preventing and treating inflammatory diseases.http://www.sciencedirect.com/science/article/pii/S2405844022012051Torin2RAW264.7 murine macrophagesLipopolysaccharideAnti-inflammationMechanisms |
| spellingShingle | Shanshan Wei Ting Wang Qi Cao Xue Chen Zhiyong Chu Zhenzhen Zhang Anti-inflammatory effects of Torin2 on lipopolysaccharide-treated RAW264.7 murine macrophages and potential mechanisms Heliyon Torin2 RAW264.7 murine macrophages Lipopolysaccharide Anti-inflammation Mechanisms |
| title | Anti-inflammatory effects of Torin2 on lipopolysaccharide-treated RAW264.7 murine macrophages and potential mechanisms |
| title_full | Anti-inflammatory effects of Torin2 on lipopolysaccharide-treated RAW264.7 murine macrophages and potential mechanisms |
| title_fullStr | Anti-inflammatory effects of Torin2 on lipopolysaccharide-treated RAW264.7 murine macrophages and potential mechanisms |
| title_full_unstemmed | Anti-inflammatory effects of Torin2 on lipopolysaccharide-treated RAW264.7 murine macrophages and potential mechanisms |
| title_short | Anti-inflammatory effects of Torin2 on lipopolysaccharide-treated RAW264.7 murine macrophages and potential mechanisms |
| title_sort | anti inflammatory effects of torin2 on lipopolysaccharide treated raw264 7 murine macrophages and potential mechanisms |
| topic | Torin2 RAW264.7 murine macrophages Lipopolysaccharide Anti-inflammation Mechanisms |
| url | http://www.sciencedirect.com/science/article/pii/S2405844022012051 |
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