Berberine Improves Glucose and Lipid Metabolism in HepG2 Cells Through AMPKα1 Activation
AimThis study is designed to investigate whether or not AMP-activated protein kinase α1 (AMPKα1) is required for natural product berberine (BBR) to improve glucose and lipid metabolism in HepG2 cells.MethodsAMPKα1 knocked-out (KO, AMPKα1-/-) cells were obtained by co-transfection of the CRISPR/Cas9...
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Frontiers Media S.A.
2020-05-01
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| Series: | Frontiers in Pharmacology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fphar.2020.00647/full |
| _version_ | 1828531612876275712 |
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| author | Gang Ren Jiang-Hong Guo Yu-Zhen Qian Yu-Zhen Qian Wei-Jia Kong Jian-Dong Jiang Jian-Dong Jiang |
| author_facet | Gang Ren Jiang-Hong Guo Yu-Zhen Qian Yu-Zhen Qian Wei-Jia Kong Jian-Dong Jiang Jian-Dong Jiang |
| author_sort | Gang Ren |
| collection | DOAJ |
| description | AimThis study is designed to investigate whether or not AMP-activated protein kinase α1 (AMPKα1) is required for natural product berberine (BBR) to improve glucose and lipid metabolism in HepG2 cells.MethodsAMPKα1 knocked-out (KO, AMPKα1-/-) cells were obtained by co-transfection of the CRISPR/Cas9 KO and HDR (homology-directed repair) plasmid into HepG2 cells, as well as subsequent screen with puromycin. The expression levels of target proteins or mRNAs were determined by western blot or real-time RT-PCR, respectively. Cellular AMPK activity, glucose consumption, lactate release, glucose production, and lipid accumulation were determined by kits.ResultsThe results showed that the AMPKα1 gene was successfully KO in HepG2 cells. In AMPKα1-/- cells, the protein expression of AMPKα1 and phosphorylated-AMPKα1 (p-AMPKα1) disappeared, the level of total AMPKα declined to about 45–50% of wild type (p < 0.01), while p-AMPKα level and AMPK activity were reduced to less than 10% of wild type (p < 0.001). BBR increased p-AMPKα1, p-AMPKα, AMPK activity, and stimulated glucose consumption, lactate release, inhibited glucose production in wild type HepG2 cells (p < 0.05 or p < 0.01). BBR also reduced intracellular lipid accumulation and suppressed the expression of lipogenic genes in oleic acid (OA) treated wild type HepG2 cells (p < 0.05 or p < 0.01). In AMPKα1-/- HepG2 cells, the stimulating effects of BBR on p-AMPKα1, p-AMPKα, AMPK activity, and its improving effects on glucose and lipid metabolism were completely abolished.ConclusionOur study proves that AMPKα1 plays a critical role for BBR to improve glucose and lipid metabolism in HepG2 cells. Our results will provide new information to further understand the molecular mechanisms of BBR. |
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| format | Article |
| id | doaj.art-3c3c6b7c8cd7432c9ac2c11e9271d00c |
| institution | Directory Open Access Journal |
| issn | 1663-9812 |
| language | English |
| last_indexed | 2024-12-11T22:40:20Z |
| publishDate | 2020-05-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Pharmacology |
| spelling | doaj.art-3c3c6b7c8cd7432c9ac2c11e9271d00c2022-12-22T00:47:49ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122020-05-011110.3389/fphar.2020.00647510652Berberine Improves Glucose and Lipid Metabolism in HepG2 Cells Through AMPKα1 ActivationGang Ren0Jiang-Hong Guo1Yu-Zhen Qian2Yu-Zhen Qian3Wei-Jia Kong4Jian-Dong Jiang5Jian-Dong Jiang6Department of Virology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, ChinaState Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, ChinaDepartment of Virology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, ChinaSchool of Life Sciences, Liaoning Normal University, Dalian, ChinaDepartment of Virology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, ChinaDepartment of Virology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, ChinaState Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, ChinaAimThis study is designed to investigate whether or not AMP-activated protein kinase α1 (AMPKα1) is required for natural product berberine (BBR) to improve glucose and lipid metabolism in HepG2 cells.MethodsAMPKα1 knocked-out (KO, AMPKα1-/-) cells were obtained by co-transfection of the CRISPR/Cas9 KO and HDR (homology-directed repair) plasmid into HepG2 cells, as well as subsequent screen with puromycin. The expression levels of target proteins or mRNAs were determined by western blot or real-time RT-PCR, respectively. Cellular AMPK activity, glucose consumption, lactate release, glucose production, and lipid accumulation were determined by kits.ResultsThe results showed that the AMPKα1 gene was successfully KO in HepG2 cells. In AMPKα1-/- cells, the protein expression of AMPKα1 and phosphorylated-AMPKα1 (p-AMPKα1) disappeared, the level of total AMPKα declined to about 45–50% of wild type (p < 0.01), while p-AMPKα level and AMPK activity were reduced to less than 10% of wild type (p < 0.001). BBR increased p-AMPKα1, p-AMPKα, AMPK activity, and stimulated glucose consumption, lactate release, inhibited glucose production in wild type HepG2 cells (p < 0.05 or p < 0.01). BBR also reduced intracellular lipid accumulation and suppressed the expression of lipogenic genes in oleic acid (OA) treated wild type HepG2 cells (p < 0.05 or p < 0.01). In AMPKα1-/- HepG2 cells, the stimulating effects of BBR on p-AMPKα1, p-AMPKα, AMPK activity, and its improving effects on glucose and lipid metabolism were completely abolished.ConclusionOur study proves that AMPKα1 plays a critical role for BBR to improve glucose and lipid metabolism in HepG2 cells. Our results will provide new information to further understand the molecular mechanisms of BBR.https://www.frontiersin.org/article/10.3389/fphar.2020.00647/fullberberineAMP-activated protein kinase α1glucose consumptiongluconeogenesislipogenesis |
| spellingShingle | Gang Ren Jiang-Hong Guo Yu-Zhen Qian Yu-Zhen Qian Wei-Jia Kong Jian-Dong Jiang Jian-Dong Jiang Berberine Improves Glucose and Lipid Metabolism in HepG2 Cells Through AMPKα1 Activation Frontiers in Pharmacology berberine AMP-activated protein kinase α1 glucose consumption gluconeogenesis lipogenesis |
| title | Berberine Improves Glucose and Lipid Metabolism in HepG2 Cells Through AMPKα1 Activation |
| title_full | Berberine Improves Glucose and Lipid Metabolism in HepG2 Cells Through AMPKα1 Activation |
| title_fullStr | Berberine Improves Glucose and Lipid Metabolism in HepG2 Cells Through AMPKα1 Activation |
| title_full_unstemmed | Berberine Improves Glucose and Lipid Metabolism in HepG2 Cells Through AMPKα1 Activation |
| title_short | Berberine Improves Glucose and Lipid Metabolism in HepG2 Cells Through AMPKα1 Activation |
| title_sort | berberine improves glucose and lipid metabolism in hepg2 cells through ampkα1 activation |
| topic | berberine AMP-activated protein kinase α1 glucose consumption gluconeogenesis lipogenesis |
| url | https://www.frontiersin.org/article/10.3389/fphar.2020.00647/full |
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