Extraction and Purification of Flavonoids from <i>Buddleja officinalis</i> Maxim and Their Attenuation of H<sub>2</sub>O<sub>2</sub>-Induced Cell Injury by Modulating Oxidative Stress and Autophagy
Cataracts are an ailment representing the leading cause of blindness in the world. The pathogenesis of cataracts is not clear, and there is no effective treatment. An increasing amount of evidence shows that oxidative stress and autophagy in lens epithelial cells play a key role in the occurrence an...
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MDPI AG
2022-12-01
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| author | Shaofeng Wei Xiaoyi Liu K. M. Faridul Hasan Yang Peng Jiao Xie Shuai Chen Qibing Zeng Peng Luo |
| author_facet | Shaofeng Wei Xiaoyi Liu K. M. Faridul Hasan Yang Peng Jiao Xie Shuai Chen Qibing Zeng Peng Luo |
| author_sort | Shaofeng Wei |
| collection | DOAJ |
| description | Cataracts are an ailment representing the leading cause of blindness in the world. The pathogenesis of cataracts is not clear, and there is no effective treatment. An increasing amount of evidence shows that oxidative stress and autophagy in lens epithelial cells play a key role in the occurrence and development of cataracts. <i>Buddleja officinalis</i> Maxim flavonoids (BMF) are natural antioxidants and regulators that present anti-inflammatory and anti-tumor effects, among others. In this study, we optimized the extraction method of BMFs and detected three of their main active monomers (luteolin, apigenin, and acacetin). In addition, a model of oxidative damage model using rabbit lens epithelial cells induced by hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>). By detecting the levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), and OH (OH), the expression of autophagosomes and autolysosomes were observed after MRFP-GFP-LC3 adenovirus was introduced into the cells. Western blotting was used to detect the expression of Beclin-1 and P62. Our research results showed that the optimal extraction parameters to obtain the highest yield of total flavonoids were a liquid–solid ratio of 1:31 g/mL, an ethanol volume fraction of 67%, an extraction time of 2.6 h, and an extraction temperature of 58 °C. Moreover, the content of luteolin was 690.85 ppb, that of apigenin was 114.91 ppb, and the content of acacetin was 5.617 ppb. After oxidative damage was induced by H<sub>2</sub>O<sub>2</sub>, the cell survival rate decreased significantly. BMFs could increase the levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) and decrease the levels of malondialdehyde (MDA) and OH (OH). After the MRFP-GFP-LC3 virus was introduced into rabbit lens epithelial cells and detecting the expression of P62 and Beclin-1, we found that the intervention of BMF could promote the binding of autophagosomes to lysosomes. Compared with the model group, the level of P62 in the low-, middle-, and high-dose groups of BMF was significantly down-regulated, the level of Beclin-1 was significantly increased, and the difference was statistically significant (<i>p</i> < 0.05). In other words, the optimized extraction method was better than others, and the purified BMF contained three main active monomers (luteolin, apigenin, and acacetin). In addition, BMFs could ameliorate the H<sub>2</sub>O<sub>2</sub>-induced oxidative damage to rabbit lens cells by promoting autophagy and regulating the level of antioxidation. |
| first_indexed | 2024-03-09T16:02:00Z |
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| spelling | doaj.art-23649fef6a7b4a3aadd320f548036ee72023-11-24T17:00:53ZengMDPI AGMolecules1420-30492022-12-012724898510.3390/molecules27248985Extraction and Purification of Flavonoids from <i>Buddleja officinalis</i> Maxim and Their Attenuation of H<sub>2</sub>O<sub>2</sub>-Induced Cell Injury by Modulating Oxidative Stress and AutophagyShaofeng Wei0Xiaoyi Liu1K. M. Faridul Hasan2Yang Peng3Jiao Xie4Shuai Chen5Qibing Zeng6Peng Luo7School of Public Health, The Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical University, Guiyang 550025, ChinaSchool of Public Health, The Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical University, Guiyang 550025, ChinaSimonyi Károly Faculty of Engineering, University of Sopron, 9400 Sopron, HungarySchool of Public Health, The Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical University, Guiyang 550025, ChinaSchool of Public Health, The Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical University, Guiyang 550025, ChinaDepartment of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Guizhou Medical University, Guiyang 550025, ChinaSchool of Public Health, The Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical University, Guiyang 550025, ChinaSchool of Public Health, The Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical University, Guiyang 550025, ChinaCataracts are an ailment representing the leading cause of blindness in the world. The pathogenesis of cataracts is not clear, and there is no effective treatment. An increasing amount of evidence shows that oxidative stress and autophagy in lens epithelial cells play a key role in the occurrence and development of cataracts. <i>Buddleja officinalis</i> Maxim flavonoids (BMF) are natural antioxidants and regulators that present anti-inflammatory and anti-tumor effects, among others. In this study, we optimized the extraction method of BMFs and detected three of their main active monomers (luteolin, apigenin, and acacetin). In addition, a model of oxidative damage model using rabbit lens epithelial cells induced by hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>). By detecting the levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), and OH (OH), the expression of autophagosomes and autolysosomes were observed after MRFP-GFP-LC3 adenovirus was introduced into the cells. Western blotting was used to detect the expression of Beclin-1 and P62. Our research results showed that the optimal extraction parameters to obtain the highest yield of total flavonoids were a liquid–solid ratio of 1:31 g/mL, an ethanol volume fraction of 67%, an extraction time of 2.6 h, and an extraction temperature of 58 °C. Moreover, the content of luteolin was 690.85 ppb, that of apigenin was 114.91 ppb, and the content of acacetin was 5.617 ppb. After oxidative damage was induced by H<sub>2</sub>O<sub>2</sub>, the cell survival rate decreased significantly. BMFs could increase the levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) and decrease the levels of malondialdehyde (MDA) and OH (OH). After the MRFP-GFP-LC3 virus was introduced into rabbit lens epithelial cells and detecting the expression of P62 and Beclin-1, we found that the intervention of BMF could promote the binding of autophagosomes to lysosomes. Compared with the model group, the level of P62 in the low-, middle-, and high-dose groups of BMF was significantly down-regulated, the level of Beclin-1 was significantly increased, and the difference was statistically significant (<i>p</i> < 0.05). In other words, the optimized extraction method was better than others, and the purified BMF contained three main active monomers (luteolin, apigenin, and acacetin). In addition, BMFs could ameliorate the H<sub>2</sub>O<sub>2</sub>-induced oxidative damage to rabbit lens cells by promoting autophagy and regulating the level of antioxidation.https://www.mdpi.com/1420-3049/27/24/8985<i>Buddleja officinalis</i> Maximflavonoidsextractionoxidative stressautophagy |
| spellingShingle | Shaofeng Wei Xiaoyi Liu K. M. Faridul Hasan Yang Peng Jiao Xie Shuai Chen Qibing Zeng Peng Luo Extraction and Purification of Flavonoids from <i>Buddleja officinalis</i> Maxim and Their Attenuation of H<sub>2</sub>O<sub>2</sub>-Induced Cell Injury by Modulating Oxidative Stress and Autophagy Molecules <i>Buddleja officinalis</i> Maxim flavonoids extraction oxidative stress autophagy |
| title | Extraction and Purification of Flavonoids from <i>Buddleja officinalis</i> Maxim and Their Attenuation of H<sub>2</sub>O<sub>2</sub>-Induced Cell Injury by Modulating Oxidative Stress and Autophagy |
| title_full | Extraction and Purification of Flavonoids from <i>Buddleja officinalis</i> Maxim and Their Attenuation of H<sub>2</sub>O<sub>2</sub>-Induced Cell Injury by Modulating Oxidative Stress and Autophagy |
| title_fullStr | Extraction and Purification of Flavonoids from <i>Buddleja officinalis</i> Maxim and Their Attenuation of H<sub>2</sub>O<sub>2</sub>-Induced Cell Injury by Modulating Oxidative Stress and Autophagy |
| title_full_unstemmed | Extraction and Purification of Flavonoids from <i>Buddleja officinalis</i> Maxim and Their Attenuation of H<sub>2</sub>O<sub>2</sub>-Induced Cell Injury by Modulating Oxidative Stress and Autophagy |
| title_short | Extraction and Purification of Flavonoids from <i>Buddleja officinalis</i> Maxim and Their Attenuation of H<sub>2</sub>O<sub>2</sub>-Induced Cell Injury by Modulating Oxidative Stress and Autophagy |
| title_sort | extraction and purification of flavonoids from i buddleja officinalis i maxim and their attenuation of h sub 2 sub o sub 2 sub induced cell injury by modulating oxidative stress and autophagy |
| topic | <i>Buddleja officinalis</i> Maxim flavonoids extraction oxidative stress autophagy |
| url | https://www.mdpi.com/1420-3049/27/24/8985 |
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