Antioxidant and anti-inflammatory function of walnut green husk aqueous extract (WNGH-AE) on human hepatocellular carcinoma cells (HepG2) treated with t-BHP.
Oxidative damage, oxidative inflammation, and a range of downstream diseases represent significant threats to human health. The application of natural antioxidants and anti-inflammatory agents can help prevent and mitigate these associated diseases. In this study, we aimed to investigate the effecti...
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Public Library of Science (PLoS)
2025-01-01
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| Online Access: | https://doi.org/10.1371/journal.pone.0318005 |
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| author | Zheng-Qiang Li Yu-Ting Gao Chen-Fa Zhao Rui An Yan-Lv Wu Zhi-Pang Huang Ping Ma Xu Yang Rong She Xiao-Yan Yang |
| author_facet | Zheng-Qiang Li Yu-Ting Gao Chen-Fa Zhao Rui An Yan-Lv Wu Zhi-Pang Huang Ping Ma Xu Yang Rong She Xiao-Yan Yang |
| author_sort | Zheng-Qiang Li |
| collection | DOAJ |
| description | Oxidative damage, oxidative inflammation, and a range of downstream diseases represent significant threats to human health. The application of natural antioxidants and anti-inflammatory agents can help prevent and mitigate these associated diseases. In this study, we aimed to investigate the effectiveness of walnut green husk (WNGH) as an antioxidant and anti-inflammatory agent in an in vitro setting. HepG2 cells were treated with tert-butyl hydroperoxide (t-BHP) to establish a cellular model of oxidative damage and inflammation. We assessed the biocompatibility of walnut green husk aqueous extract (WNGH-AE) on HepG2 cells using MTT and LDH assays (WNGH-AE concentration: 0.05, 0.1, 0.2, 0.4, 0.8, 1.6, 3.2, 6.4, and 12.8 mg/mL). Additionally, we measured intracellular oxidative stress indicators, such as ROS and 8-OHdG, along with inflammatory factors TNF-α and IL-1β through ELISA to evaluate the antioxidant and anti-inflammatory capacity of WNGH-AE (concentration: 0.025, 0.05, 0.1 mg/mL) in HepG2 cells. We also determined the free radical scavenging capacity of various extracts of WNGH using DPPH and ·OH methods. The total phenols, total polysaccharides, and total flavonoids in WNGH-AE were analyzed using the Folin-Ciocalteu's reagent, the phenol-sulfuric acid method, and the spectrophotometry, respectively. The bioactive components of WNGH-AE were analyzed using LC-MS/MS. Our results demonstrated that WNGH-AE was highly biocompatible with HepG2 cells. The antioxidant effect of WNGH-AE involved the scavenging of intracellular ROS, while its anti-inflammatory effect was linked to the down-regulation of the NF-κB pathway. Compared to other extractants (ethyl acetate, n-butanol, 75% ethanol, and petroleum ether), WNGH-AE exhibited the strongest free radical scavenging ability. Through LC-MS/MS analysis, we identified 403 compounds in WNGH-AE, with gentisic acid being the most abundant and possessing high antioxidant capacity, suggesting it may be a key active component contributing to the antioxidant activity of WNGH-AE. In conclusion, our findings indicate that WNGH-AE is a natural, high-quality antioxidant and anti-inflammatory biomaterial deserving further research and development, with significant potential applications in healthcare. (311 words). |
| format | Article |
| id | doaj-art-cf7cfffc2c8d4284a373c73af2585d95 |
| institution | Kabale University |
| issn | 1932-6203 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Public Library of Science (PLoS) |
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| spelling | doaj-art-cf7cfffc2c8d4284a373c73af2585d952025-02-05T05:32:00ZengPublic Library of Science (PLoS)PLoS ONE1932-62032025-01-01201e031800510.1371/journal.pone.0318005Antioxidant and anti-inflammatory function of walnut green husk aqueous extract (WNGH-AE) on human hepatocellular carcinoma cells (HepG2) treated with t-BHP.Zheng-Qiang LiYu-Ting GaoChen-Fa ZhaoRui AnYan-Lv WuZhi-Pang HuangPing MaXu YangRong SheXiao-Yan YangOxidative damage, oxidative inflammation, and a range of downstream diseases represent significant threats to human health. The application of natural antioxidants and anti-inflammatory agents can help prevent and mitigate these associated diseases. In this study, we aimed to investigate the effectiveness of walnut green husk (WNGH) as an antioxidant and anti-inflammatory agent in an in vitro setting. HepG2 cells were treated with tert-butyl hydroperoxide (t-BHP) to establish a cellular model of oxidative damage and inflammation. We assessed the biocompatibility of walnut green husk aqueous extract (WNGH-AE) on HepG2 cells using MTT and LDH assays (WNGH-AE concentration: 0.05, 0.1, 0.2, 0.4, 0.8, 1.6, 3.2, 6.4, and 12.8 mg/mL). Additionally, we measured intracellular oxidative stress indicators, such as ROS and 8-OHdG, along with inflammatory factors TNF-α and IL-1β through ELISA to evaluate the antioxidant and anti-inflammatory capacity of WNGH-AE (concentration: 0.025, 0.05, 0.1 mg/mL) in HepG2 cells. We also determined the free radical scavenging capacity of various extracts of WNGH using DPPH and ·OH methods. The total phenols, total polysaccharides, and total flavonoids in WNGH-AE were analyzed using the Folin-Ciocalteu's reagent, the phenol-sulfuric acid method, and the spectrophotometry, respectively. The bioactive components of WNGH-AE were analyzed using LC-MS/MS. Our results demonstrated that WNGH-AE was highly biocompatible with HepG2 cells. The antioxidant effect of WNGH-AE involved the scavenging of intracellular ROS, while its anti-inflammatory effect was linked to the down-regulation of the NF-κB pathway. Compared to other extractants (ethyl acetate, n-butanol, 75% ethanol, and petroleum ether), WNGH-AE exhibited the strongest free radical scavenging ability. Through LC-MS/MS analysis, we identified 403 compounds in WNGH-AE, with gentisic acid being the most abundant and possessing high antioxidant capacity, suggesting it may be a key active component contributing to the antioxidant activity of WNGH-AE. In conclusion, our findings indicate that WNGH-AE is a natural, high-quality antioxidant and anti-inflammatory biomaterial deserving further research and development, with significant potential applications in healthcare. (311 words).https://doi.org/10.1371/journal.pone.0318005 |
| spellingShingle | Zheng-Qiang Li Yu-Ting Gao Chen-Fa Zhao Rui An Yan-Lv Wu Zhi-Pang Huang Ping Ma Xu Yang Rong She Xiao-Yan Yang Antioxidant and anti-inflammatory function of walnut green husk aqueous extract (WNGH-AE) on human hepatocellular carcinoma cells (HepG2) treated with t-BHP. PLoS ONE |
| title | Antioxidant and anti-inflammatory function of walnut green husk aqueous extract (WNGH-AE) on human hepatocellular carcinoma cells (HepG2) treated with t-BHP. |
| title_full | Antioxidant and anti-inflammatory function of walnut green husk aqueous extract (WNGH-AE) on human hepatocellular carcinoma cells (HepG2) treated with t-BHP. |
| title_fullStr | Antioxidant and anti-inflammatory function of walnut green husk aqueous extract (WNGH-AE) on human hepatocellular carcinoma cells (HepG2) treated with t-BHP. |
| title_full_unstemmed | Antioxidant and anti-inflammatory function of walnut green husk aqueous extract (WNGH-AE) on human hepatocellular carcinoma cells (HepG2) treated with t-BHP. |
| title_short | Antioxidant and anti-inflammatory function of walnut green husk aqueous extract (WNGH-AE) on human hepatocellular carcinoma cells (HepG2) treated with t-BHP. |
| title_sort | antioxidant and anti inflammatory function of walnut green husk aqueous extract wngh ae on human hepatocellular carcinoma cells hepg2 treated with t bhp |
| url | https://doi.org/10.1371/journal.pone.0318005 |
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