Investigating the Anti-Inflammatory Effects of RCI001 for Treating Ocular Surface Diseases: Insight Into the Mechanism of Action
The ocular surface is continuously exposed to various environmental factors, and innate and adaptive immunity play crucial roles in ocular surface diseases (OSDs). Previously, we have reported that the topical application of RCI001 affords excellent anti-inflammatory and antioxidant effects in dry e...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-03-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2022.850287/full |
| _version_ | 1818776742584647680 |
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| author | Seunghoon Kim Seunghoon Kim Ye Won Jang Young-ah Ku Yungyeong Shin Md Mahbubur Rahman Myung-Hee Chung Yong Ho Kim Yong Ho Kim Dong Hyun Kim Dong Hyun Kim |
| author_facet | Seunghoon Kim Seunghoon Kim Ye Won Jang Young-ah Ku Yungyeong Shin Md Mahbubur Rahman Myung-Hee Chung Yong Ho Kim Yong Ho Kim Dong Hyun Kim Dong Hyun Kim |
| author_sort | Seunghoon Kim |
| collection | DOAJ |
| description | The ocular surface is continuously exposed to various environmental factors, and innate and adaptive immunity play crucial roles in ocular surface diseases (OSDs). Previously, we have reported that the topical application of RCI001 affords excellent anti-inflammatory and antioxidant effects in dry eye disease and ocular chemical burn models. In this study, we examined the inhibitory effects of RCI001 on the Rac1 and NLRP3 inflammasomes in vitro and in vivo. Following RCI001 application to RAW264.7 and Swiss 3T3 cells, we measured Rac1 activity using a glutathione-S-transferase (GST) pull-down assay and G-protein activation assay kit. In addition, we quantified the expression of inflammatory cytokines (interleukin [IL]-1β, IL-6, and tumor necrosis factor [TNF]-α) in lipopolysaccharide (LPS)-stimulated RAW264.7 cells using ELISA and real-time PCR. In the mouse ocular alkali burn model, RCI001 was administered via eye drops (10 mg/mL, twice daily) for 5 days, and 1% prednisolone acetate (PDE) ophthalmic suspension was used as a positive control. Corneal epithelial integrity (on days 0-5) and histological examinations were performed, and transcript and protein levels of Rac1, NLRP3, caspase-1, and IL-1β were quantified using real-time PCR and western blotting in corneal tissues collected on days 3 and 5. We observed that RCI001 dose-dependently inhibited Rac1 activity and various inflammatory cytokines in LPS-stimulated murine macrophages. Furthermore, RCI001 restored corneal epithelial integrity more rapidly than corticosteroid treatment in chemically injured corneas. Compared to the saline group, activation of Rac1 and the NLRP3 inflammasome/IL-1β axis was suppressed in the RCI001 group, especially during the early phase of the ocular alkali burn model. Topical RCI001 suppressed the expression of activated Rac1 and inflammatory cytokines in vitro and rapidly restored the injured cornea by inhibiting activation of Rac1 and the NLRP inflammasome/IL-1β axis in vivo. Accordingly, RCI001 could be a promising therapeutic agent for treating OSDs. |
| first_indexed | 2024-12-18T11:17:46Z |
| format | Article |
| id | doaj.art-3a523164942448928af3fdb190976b63 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-18T11:17:46Z |
| publishDate | 2022-03-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Immunology |
| spelling | doaj.art-3a523164942448928af3fdb190976b632022-12-21T21:09:54ZengFrontiers Media S.A.Frontiers in Immunology1664-32242022-03-011310.3389/fimmu.2022.850287850287Investigating the Anti-Inflammatory Effects of RCI001 for Treating Ocular Surface Diseases: Insight Into the Mechanism of ActionSeunghoon Kim0Seunghoon Kim1Ye Won Jang2Young-ah Ku3Yungyeong Shin4Md Mahbubur Rahman5Myung-Hee Chung6Yong Ho Kim7Yong Ho Kim8Dong Hyun Kim9Dong Hyun Kim10RudaCure Co. Ltd., Incheon, South KoreaGachon Pain Center and Department of Physiology, Gachon University College of Medicine, Incheon, South KoreaRudaCure Co. Ltd., Incheon, South KoreaRudaCure Co. Ltd., Incheon, South KoreaGachon Pain Center and Department of Physiology, Gachon University College of Medicine, Incheon, South KoreaGachon Pain Center and Department of Physiology, Gachon University College of Medicine, Incheon, South KoreaRudaCure Co. Ltd., Incheon, South KoreaRudaCure Co. Ltd., Incheon, South KoreaGachon Pain Center and Department of Physiology, Gachon University College of Medicine, Incheon, South KoreaRudaCure Co. Ltd., Incheon, South KoreaDepartment of Ophthalmology, Gil Medical Center, Gachon University College of Medicine, Incheon, South KoreaThe ocular surface is continuously exposed to various environmental factors, and innate and adaptive immunity play crucial roles in ocular surface diseases (OSDs). Previously, we have reported that the topical application of RCI001 affords excellent anti-inflammatory and antioxidant effects in dry eye disease and ocular chemical burn models. In this study, we examined the inhibitory effects of RCI001 on the Rac1 and NLRP3 inflammasomes in vitro and in vivo. Following RCI001 application to RAW264.7 and Swiss 3T3 cells, we measured Rac1 activity using a glutathione-S-transferase (GST) pull-down assay and G-protein activation assay kit. In addition, we quantified the expression of inflammatory cytokines (interleukin [IL]-1β, IL-6, and tumor necrosis factor [TNF]-α) in lipopolysaccharide (LPS)-stimulated RAW264.7 cells using ELISA and real-time PCR. In the mouse ocular alkali burn model, RCI001 was administered via eye drops (10 mg/mL, twice daily) for 5 days, and 1% prednisolone acetate (PDE) ophthalmic suspension was used as a positive control. Corneal epithelial integrity (on days 0-5) and histological examinations were performed, and transcript and protein levels of Rac1, NLRP3, caspase-1, and IL-1β were quantified using real-time PCR and western blotting in corneal tissues collected on days 3 and 5. We observed that RCI001 dose-dependently inhibited Rac1 activity and various inflammatory cytokines in LPS-stimulated murine macrophages. Furthermore, RCI001 restored corneal epithelial integrity more rapidly than corticosteroid treatment in chemically injured corneas. Compared to the saline group, activation of Rac1 and the NLRP3 inflammasome/IL-1β axis was suppressed in the RCI001 group, especially during the early phase of the ocular alkali burn model. Topical RCI001 suppressed the expression of activated Rac1 and inflammatory cytokines in vitro and rapidly restored the injured cornea by inhibiting activation of Rac1 and the NLRP inflammasome/IL-1β axis in vivo. Accordingly, RCI001 could be a promising therapeutic agent for treating OSDs.https://www.frontiersin.org/articles/10.3389/fimmu.2022.850287/fullRCI001ocular surface disease (OSD)inflammationRac1NLRP3 inflammasome |
| spellingShingle | Seunghoon Kim Seunghoon Kim Ye Won Jang Young-ah Ku Yungyeong Shin Md Mahbubur Rahman Myung-Hee Chung Yong Ho Kim Yong Ho Kim Dong Hyun Kim Dong Hyun Kim Investigating the Anti-Inflammatory Effects of RCI001 for Treating Ocular Surface Diseases: Insight Into the Mechanism of Action Frontiers in Immunology RCI001 ocular surface disease (OSD) inflammation Rac1 NLRP3 inflammasome |
| title | Investigating the Anti-Inflammatory Effects of RCI001 for Treating Ocular Surface Diseases: Insight Into the Mechanism of Action |
| title_full | Investigating the Anti-Inflammatory Effects of RCI001 for Treating Ocular Surface Diseases: Insight Into the Mechanism of Action |
| title_fullStr | Investigating the Anti-Inflammatory Effects of RCI001 for Treating Ocular Surface Diseases: Insight Into the Mechanism of Action |
| title_full_unstemmed | Investigating the Anti-Inflammatory Effects of RCI001 for Treating Ocular Surface Diseases: Insight Into the Mechanism of Action |
| title_short | Investigating the Anti-Inflammatory Effects of RCI001 for Treating Ocular Surface Diseases: Insight Into the Mechanism of Action |
| title_sort | investigating the anti inflammatory effects of rci001 for treating ocular surface diseases insight into the mechanism of action |
| topic | RCI001 ocular surface disease (OSD) inflammation Rac1 NLRP3 inflammasome |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2022.850287/full |
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