Anti-inflammatory and antioxidative effects of gallic acid on experimental dry eye: in vitro and in vivo studies

Abstract Background To investigate the anti-inflammatory and antioxidative effects of gallic acid (GA) on human corneal epithelial cells (HCECs) and RAW264.7 macrophages as well as its therapeutic effects in an experimental dry eye (EDE) mouse model. Methods A cell counting kit-8 (CCK-8) assay was u...

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Main Authors: Kexin Li, Qianwen Gong, Bin Lu, Kaiyan Huang, Yixuan Tong, Tinashe Emmanuel Mutsvene, Meng Lin, Zhiqiang Xu, Fan Lu, Xingyi Li, Liang Hu
Format: Article
Language:English
Published: BMC 2023-05-01
Series:Eye and Vision
Subjects:
Online Access:https://doi.org/10.1186/s40662-023-00334-5
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author Kexin Li
Qianwen Gong
Bin Lu
Kaiyan Huang
Yixuan Tong
Tinashe Emmanuel Mutsvene
Meng Lin
Zhiqiang Xu
Fan Lu
Xingyi Li
Liang Hu
author_facet Kexin Li
Qianwen Gong
Bin Lu
Kaiyan Huang
Yixuan Tong
Tinashe Emmanuel Mutsvene
Meng Lin
Zhiqiang Xu
Fan Lu
Xingyi Li
Liang Hu
author_sort Kexin Li
collection DOAJ
description Abstract Background To investigate the anti-inflammatory and antioxidative effects of gallic acid (GA) on human corneal epithelial cells (HCECs) and RAW264.7 macrophages as well as its therapeutic effects in an experimental dry eye (EDE) mouse model. Methods A cell counting kit-8 (CCK-8) assay was used to test the cytotoxicity of GA. The effect of GA on cell migration was evaluated using a scratch wound healing assay. The anti-inflammatory and antioxidative effects of GA in vitro were tested using a hypertonic model (HCECs) and an inflammatory model (RAW264.7 cells). The in vivo biocompatibility of GA was detected by irritation tests in rabbits, whereas the preventive and therapeutic effect of GA in vivo was evaluated using a mouse model of EDE. Results In the range of 0–100 μM, GA showed no cytotoxicity in RAW264.7 cells or HCECs and did not delay the HCECs monolayer wound healing within 24 h. Ocular tolerance to GA in the in vivo irritation test was good after seven days. In terms of antioxidative activity, GA significantly reduced the intracellular reactive oxygen species (ROS) in lipopolysaccharide (LPS) activated RAW264.7 macrophages and HCECs exposed to hyperosmotic stress. Furthermore, after pre-treatment with GA, the expression levels of nuclear factor E2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and NADPH quinone oxidoreductase-1 (NQO-1) were significantly upregulated in RAW264.7 macrophages. GA also exhibits excellent anti-inflammatory properties. This is mainly demonstrated by the ability of GA to effectively downregulate the nuclear transcription factor-κB (NF-κB) pathway in LPS-activated RAW264.7 macrophages and to reduce inflammatory factors, such as nitric oxide (NO), interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-α). In vivo efficacy testing results in a mouse model of EDE showed that GA can effectively prevent and inhibit the apoptosis of corneal epithelial cells (CECs), reduce inflammatory factors in the cornea and conjunctiva as well as protect goblet cells. Conclusion In vitro and in vivo results indicate that GA possesses potent anti-inflammatory and antioxidative properties with no apparent cytotoxicity within the range of 0–100 μM. It is a promising eye drop formulation for the effective prevention and treatment of dry eye disease (DED).
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spelling doaj.art-921687cb775d4dbf8000bfedacb077392023-05-07T11:05:45ZengBMCEye and Vision2326-02542023-05-0110111710.1186/s40662-023-00334-5Anti-inflammatory and antioxidative effects of gallic acid on experimental dry eye: in vitro and in vivo studiesKexin Li0Qianwen Gong1Bin Lu2Kaiyan Huang3Yixuan Tong4Tinashe Emmanuel Mutsvene5Meng Lin6Zhiqiang Xu7Fan Lu8Xingyi Li9Liang Hu10National Clinical Research Center for Ocular Diseases, Eye Hospital, Wenzhou Medical UniversityNational Clinical Research Center for Ocular Diseases, Eye Hospital, Wenzhou Medical UniversityNational Clinical Research Center for Ocular Diseases, Eye Hospital, Wenzhou Medical UniversityNational Clinical Research Center for Ocular Diseases, Eye Hospital, Wenzhou Medical UniversityNational Clinical Research Center for Ocular Diseases, Eye Hospital, Wenzhou Medical UniversityNational Clinical Research Center for Ocular Diseases, Eye Hospital, Wenzhou Medical UniversityNational Clinical Research Center for Ocular Diseases, Eye Hospital, Wenzhou Medical UniversityNational Clinical Research Center for Ocular Diseases, Eye Hospital, Wenzhou Medical UniversityNational Clinical Research Center for Ocular Diseases, Eye Hospital, Wenzhou Medical UniversityInstitute of Biomedical Engineering, School of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical UniversityNational Clinical Research Center for Ocular Diseases, Eye Hospital, Wenzhou Medical UniversityAbstract Background To investigate the anti-inflammatory and antioxidative effects of gallic acid (GA) on human corneal epithelial cells (HCECs) and RAW264.7 macrophages as well as its therapeutic effects in an experimental dry eye (EDE) mouse model. Methods A cell counting kit-8 (CCK-8) assay was used to test the cytotoxicity of GA. The effect of GA on cell migration was evaluated using a scratch wound healing assay. The anti-inflammatory and antioxidative effects of GA in vitro were tested using a hypertonic model (HCECs) and an inflammatory model (RAW264.7 cells). The in vivo biocompatibility of GA was detected by irritation tests in rabbits, whereas the preventive and therapeutic effect of GA in vivo was evaluated using a mouse model of EDE. Results In the range of 0–100 μM, GA showed no cytotoxicity in RAW264.7 cells or HCECs and did not delay the HCECs monolayer wound healing within 24 h. Ocular tolerance to GA in the in vivo irritation test was good after seven days. In terms of antioxidative activity, GA significantly reduced the intracellular reactive oxygen species (ROS) in lipopolysaccharide (LPS) activated RAW264.7 macrophages and HCECs exposed to hyperosmotic stress. Furthermore, after pre-treatment with GA, the expression levels of nuclear factor E2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and NADPH quinone oxidoreductase-1 (NQO-1) were significantly upregulated in RAW264.7 macrophages. GA also exhibits excellent anti-inflammatory properties. This is mainly demonstrated by the ability of GA to effectively downregulate the nuclear transcription factor-κB (NF-κB) pathway in LPS-activated RAW264.7 macrophages and to reduce inflammatory factors, such as nitric oxide (NO), interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-α). In vivo efficacy testing results in a mouse model of EDE showed that GA can effectively prevent and inhibit the apoptosis of corneal epithelial cells (CECs), reduce inflammatory factors in the cornea and conjunctiva as well as protect goblet cells. Conclusion In vitro and in vivo results indicate that GA possesses potent anti-inflammatory and antioxidative properties with no apparent cytotoxicity within the range of 0–100 μM. It is a promising eye drop formulation for the effective prevention and treatment of dry eye disease (DED).https://doi.org/10.1186/s40662-023-00334-5Corneal epithelial cellsMacrophagesGallic acidDry eyeInflammationOxidative stress
spellingShingle Kexin Li
Qianwen Gong
Bin Lu
Kaiyan Huang
Yixuan Tong
Tinashe Emmanuel Mutsvene
Meng Lin
Zhiqiang Xu
Fan Lu
Xingyi Li
Liang Hu
Anti-inflammatory and antioxidative effects of gallic acid on experimental dry eye: in vitro and in vivo studies
Eye and Vision
Corneal epithelial cells
Macrophages
Gallic acid
Dry eye
Inflammation
Oxidative stress
title Anti-inflammatory and antioxidative effects of gallic acid on experimental dry eye: in vitro and in vivo studies
title_full Anti-inflammatory and antioxidative effects of gallic acid on experimental dry eye: in vitro and in vivo studies
title_fullStr Anti-inflammatory and antioxidative effects of gallic acid on experimental dry eye: in vitro and in vivo studies
title_full_unstemmed Anti-inflammatory and antioxidative effects of gallic acid on experimental dry eye: in vitro and in vivo studies
title_short Anti-inflammatory and antioxidative effects of gallic acid on experimental dry eye: in vitro and in vivo studies
title_sort anti inflammatory and antioxidative effects of gallic acid on experimental dry eye in vitro and in vivo studies
topic Corneal epithelial cells
Macrophages
Gallic acid
Dry eye
Inflammation
Oxidative stress
url https://doi.org/10.1186/s40662-023-00334-5
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