Journal: Chinese Medicine

Article Title: The effect and mechanism of Germacrone in ameliorating alcoholic fatty liver by inhibiting Nrf2/Rbp4

doi: 10.1186/s13020-025-01132-y

Figure Lengend Snippet: Germacrone improves lipid accumulation and oxidative stress in a mouse model of ALD. A H&E staining revealed pathological changes in the mouse liver; Oil red O staining showed lipid accumulation in hepatocytes. B – I biochemical kit was used to detect ALT, AST, TG, HDL-C, LDL-C, GSH, SOD, and MDA levels in liver tissues (n = 6). Compared with the model group, *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: After 3 days of adaptive feeding, C57BL/6 male mice were randomly divided into the following 4 groups with 6 mice in each group: (1) Control group: Mice were fed the Lieber–DeCarli standard liquid diet for 35 consecutive days without any other intervention. (2) Model group: mice were fed a liquid diet with low alcohol concentration for the first 6 days as a transition and then changed to the Lieber–DeCarli alcoholic liquid diet with 5% alcohol concentration from the 7 th day for 28 days without drug intervention. (3) Germacrone group: The feed was treated the same as the model group, and Germacrone (molecular formula: C15H22O, purity > 98%, Yuanye, Shanghai, China, 10 mg/kg) [ , ] was given daily intragastric administration from day 8 for 21 days. (4) Brusatol group: the feed treatment was the same as the model group, and the Nrf2 antagonist Brusatol (Purity: 99.96%, Selleck, S7956, Shanghai, China, 2 mg/kg) [ ] was intraperitoneally injected every other day from the 8 th day for 21 days.

Techniques: Staining

Journal: Chinese Medicine

Article Title: The effect and mechanism of Germacrone in ameliorating alcoholic fatty liver by inhibiting Nrf2/Rbp4

doi: 10.1186/s13020-025-01132-y

Figure Lengend Snippet: Germacrone improves alcohol-induced lipid accumulation and oxidative stress in HepG2 cells. A The effect of different concentration gradients of Germacrone on HepG2 cell activity was detected by CCK-8 assay (n = 5); B – G A biochemical kit was used to detect the levels of ALT, AST, TG, GSH, SOD, and MDA in the Control group, Model group, Germacrone-L group, Germacrone-M group, and Germacrone-H group (n = 4–6). Compared with the Model group, *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: After 3 days of adaptive feeding, C57BL/6 male mice were randomly divided into the following 4 groups with 6 mice in each group: (1) Control group: Mice were fed the Lieber–DeCarli standard liquid diet for 35 consecutive days without any other intervention. (2) Model group: mice were fed a liquid diet with low alcohol concentration for the first 6 days as a transition and then changed to the Lieber–DeCarli alcoholic liquid diet with 5% alcohol concentration from the 7 th day for 28 days without drug intervention. (3) Germacrone group: The feed was treated the same as the model group, and Germacrone (molecular formula: C15H22O, purity > 98%, Yuanye, Shanghai, China, 10 mg/kg) [ , ] was given daily intragastric administration from day 8 for 21 days. (4) Brusatol group: the feed treatment was the same as the model group, and the Nrf2 antagonist Brusatol (Purity: 99.96%, Selleck, S7956, Shanghai, China, 2 mg/kg) [ ] was intraperitoneally injected every other day from the 8 th day for 21 days.

Techniques: Concentration Assay, Activity Assay, CCK-8 Assay, Control

Journal: Chinese Medicine

Article Title: The effect and mechanism of Germacrone in ameliorating alcoholic fatty liver by inhibiting Nrf2/Rbp4

doi: 10.1186/s13020-025-01132-y

Figure Lengend Snippet: Germacrone regulates Nrf2 nuclear translocation and downstream target gene expression in ALD. A – D Western blot analysis of Nrf2, HO-1, and Gsta1 protein expression levels in liver tissues of the Control group, Model group, Brusatol group and Germacrone group (n = 3). E , F The mRNA levels of HO-1 and Gsta1 in liver tissues of the Control group, Model group, Brusatol group, and Germacrone group were detected by RT-PCR (n = 6). G , H Western blot analysis showed the expression level of Nrf2 protein in each group (n = 3). I Immunofluorescence was used to observe the localization and expression of Nrf2 in HepG2 cells (magnification 200×, n = 3). J , K Western blot analysis showed HO-1 protein expression levels under different treatments (n = 3). Compared with the model group, *P < 0.05, **P < 0.01, ***P < 0.001; #P < 0.05, ##P < 0.01, ###P < 0.001 compared with the TBHQ group

Article Snippet: After 3 days of adaptive feeding, C57BL/6 male mice were randomly divided into the following 4 groups with 6 mice in each group: (1) Control group: Mice were fed the Lieber–DeCarli standard liquid diet for 35 consecutive days without any other intervention. (2) Model group: mice were fed a liquid diet with low alcohol concentration for the first 6 days as a transition and then changed to the Lieber–DeCarli alcoholic liquid diet with 5% alcohol concentration from the 7 th day for 28 days without drug intervention. (3) Germacrone group: The feed was treated the same as the model group, and Germacrone (molecular formula: C15H22O, purity > 98%, Yuanye, Shanghai, China, 10 mg/kg) [ , ] was given daily intragastric administration from day 8 for 21 days. (4) Brusatol group: the feed treatment was the same as the model group, and the Nrf2 antagonist Brusatol (Purity: 99.96%, Selleck, S7956, Shanghai, China, 2 mg/kg) [ ] was intraperitoneally injected every other day from the 8 th day for 21 days.

Techniques: Translocation Assay, Targeted Gene Expression, Western Blot, Expressing, Control, Reverse Transcription Polymerase Chain Reaction, Immunofluorescence

Journal: Chinese Medicine

Article Title: The effect and mechanism of Germacrone in ameliorating alcoholic fatty liver by inhibiting Nrf2/Rbp4

doi: 10.1186/s13020-025-01132-y

Figure Lengend Snippet: Germacrone inhibits Nrf2-dependent Rbp4 expression in the ALD model. A , B The protein expression levels of Rbp4 in the ALD model treated with Germacrone and Brusatol (n = 3). C Correlation analysis of liver TG content and Rbp4 expression. D Rbp4 mRNA expression in HepG2 cells in the Control group, Model group, Brusatol group, and Germacrone group was analyzed by RT-qPCR (n = 6). E , F Western blot was used to detect the expression level of Rbp4 protein under different conditions (n = 3). Compared with the model group, *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: After 3 days of adaptive feeding, C57BL/6 male mice were randomly divided into the following 4 groups with 6 mice in each group: (1) Control group: Mice were fed the Lieber–DeCarli standard liquid diet for 35 consecutive days without any other intervention. (2) Model group: mice were fed a liquid diet with low alcohol concentration for the first 6 days as a transition and then changed to the Lieber–DeCarli alcoholic liquid diet with 5% alcohol concentration from the 7 th day for 28 days without drug intervention. (3) Germacrone group: The feed was treated the same as the model group, and Germacrone (molecular formula: C15H22O, purity > 98%, Yuanye, Shanghai, China, 10 mg/kg) [ , ] was given daily intragastric administration from day 8 for 21 days. (4) Brusatol group: the feed treatment was the same as the model group, and the Nrf2 antagonist Brusatol (Purity: 99.96%, Selleck, S7956, Shanghai, China, 2 mg/kg) [ ] was intraperitoneally injected every other day from the 8 th day for 21 days.

Techniques: Expressing, Control, Quantitative RT-PCR, Western Blot

Journal: Chinese Medicine

Article Title: The effect and mechanism of Germacrone in ameliorating alcoholic fatty liver by inhibiting Nrf2/Rbp4

doi: 10.1186/s13020-025-01132-y

Figure Lengend Snippet: Mechanism of Germacrone in the Treatment of Alcohol-Induced Liver Disease (ALD). Under alcohol exposure, reactive oxygen species (ROS) and foreign electrophilic reagents inhibit the ubiquitination of Nrf2, leading to its nuclear translocation. In the nucleus, Nrf2 binds to antioxidant response elements (AREs), promoting the transcription of downstream target genes, including Rbp4 (lipid transport protein), HO-1, and Gsta1 (oxidative stress-regulated proteins). This process accelerates lipid transport, lipid accumulation, and oxidative damage in hepatocytes, which contributes to the progression of ALD (left panel). In contrast, Germacrone (Germacrone), the active component of Jia-Ga-Song-Tang (JGST), effectively inhibits Nrf2 activation and nuclear translocation, thereby down-regulating the expression of Rbp4, HO-1, and Gsta1. This reduces lipid accumulation and oxidative stress, protecting hepatocytes and improving liver health (right panel)

Article Snippet: After 3 days of adaptive feeding, C57BL/6 male mice were randomly divided into the following 4 groups with 6 mice in each group: (1) Control group: Mice were fed the Lieber–DeCarli standard liquid diet for 35 consecutive days without any other intervention. (2) Model group: mice were fed a liquid diet with low alcohol concentration for the first 6 days as a transition and then changed to the Lieber–DeCarli alcoholic liquid diet with 5% alcohol concentration from the 7 th day for 28 days without drug intervention. (3) Germacrone group: The feed was treated the same as the model group, and Germacrone (molecular formula: C15H22O, purity > 98%, Yuanye, Shanghai, China, 10 mg/kg) [ , ] was given daily intragastric administration from day 8 for 21 days. (4) Brusatol group: the feed treatment was the same as the model group, and the Nrf2 antagonist Brusatol (Purity: 99.96%, Selleck, S7956, Shanghai, China, 2 mg/kg) [ ] was intraperitoneally injected every other day from the 8 th day for 21 days.

Techniques: Ubiquitin Proteomics, Translocation Assay, Activation Assay, Expressing

Journal: Chinese Medicine

Article Title: The effect and mechanism of Germacrone in ameliorating alcoholic fatty liver by inhibiting Nrf2/Rbp4

doi: 10.1186/s13020-025-01132-y

Figure Lengend Snippet: Germacrone improves lipid accumulation and oxidative stress in a mouse model of ALD. A H&E staining revealed pathological changes in the mouse liver; Oil red O staining showed lipid accumulation in hepatocytes. B – I biochemical kit was used to detect ALT, AST, TG, HDL-C, LDL-C, GSH, SOD, and MDA levels in liver tissues (n = 6). Compared with the model group, *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: After 3 days of adaptive feeding, C57BL/6 male mice were randomly divided into the following 4 groups with 6 mice in each group: (1) Control group: Mice were fed the Lieber–DeCarli standard liquid diet for 35 consecutive days without any other intervention. (2) Model group: mice were fed a liquid diet with low alcohol concentration for the first 6 days as a transition and then changed to the Lieber–DeCarli alcoholic liquid diet with 5% alcohol concentration from the 7 th day for 28 days without drug intervention. (3) Germacrone group: The feed was treated the same as the model group, and Germacrone (molecular formula: C15H22O, purity > 98%, Yuanye, Shanghai, China, 10 mg/kg) [ , ] was given daily intragastric administration from day 8 for 21 days. (4) Brusatol group: the feed treatment was the same as the model group, and the Nrf2 antagonist Brusatol (Purity: 99.96%, Selleck, S7956, Shanghai, China, 2 mg/kg) [ ] was intraperitoneally injected every other day from the 8 th day for 21 days.

Techniques: Staining

Journal: Chinese Medicine

Article Title: The effect and mechanism of Germacrone in ameliorating alcoholic fatty liver by inhibiting Nrf2/Rbp4

doi: 10.1186/s13020-025-01132-y

Figure Lengend Snippet: Germacrone improves alcohol-induced lipid accumulation and oxidative stress in HepG2 cells. A The effect of different concentration gradients of Germacrone on HepG2 cell activity was detected by CCK-8 assay (n = 5); B – G A biochemical kit was used to detect the levels of ALT, AST, TG, GSH, SOD, and MDA in the Control group, Model group, Germacrone-L group, Germacrone-M group, and Germacrone-H group (n = 4–6). Compared with the Model group, *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: After 3 days of adaptive feeding, C57BL/6 male mice were randomly divided into the following 4 groups with 6 mice in each group: (1) Control group: Mice were fed the Lieber–DeCarli standard liquid diet for 35 consecutive days without any other intervention. (2) Model group: mice were fed a liquid diet with low alcohol concentration for the first 6 days as a transition and then changed to the Lieber–DeCarli alcoholic liquid diet with 5% alcohol concentration from the 7 th day for 28 days without drug intervention. (3) Germacrone group: The feed was treated the same as the model group, and Germacrone (molecular formula: C15H22O, purity > 98%, Yuanye, Shanghai, China, 10 mg/kg) [ , ] was given daily intragastric administration from day 8 for 21 days. (4) Brusatol group: the feed treatment was the same as the model group, and the Nrf2 antagonist Brusatol (Purity: 99.96%, Selleck, S7956, Shanghai, China, 2 mg/kg) [ ] was intraperitoneally injected every other day from the 8 th day for 21 days.

Techniques: Concentration Assay, Activity Assay, CCK-8 Assay, Control

Journal: Chinese Medicine

Article Title: The effect and mechanism of Germacrone in ameliorating alcoholic fatty liver by inhibiting Nrf2/Rbp4

doi: 10.1186/s13020-025-01132-y

Figure Lengend Snippet: Germacrone regulates Nrf2 nuclear translocation and downstream target gene expression in ALD. A – D Western blot analysis of Nrf2, HO-1, and Gsta1 protein expression levels in liver tissues of the Control group, Model group, Brusatol group and Germacrone group (n = 3). E , F The mRNA levels of HO-1 and Gsta1 in liver tissues of the Control group, Model group, Brusatol group, and Germacrone group were detected by RT-PCR (n = 6). G , H Western blot analysis showed the expression level of Nrf2 protein in each group (n = 3). I Immunofluorescence was used to observe the localization and expression of Nrf2 in HepG2 cells (magnification 200×, n = 3). J , K Western blot analysis showed HO-1 protein expression levels under different treatments (n = 3). Compared with the model group, *P < 0.05, **P < 0.01, ***P < 0.001; #P < 0.05, ##P < 0.01, ###P < 0.001 compared with the TBHQ group

Article Snippet: After 3 days of adaptive feeding, C57BL/6 male mice were randomly divided into the following 4 groups with 6 mice in each group: (1) Control group: Mice were fed the Lieber–DeCarli standard liquid diet for 35 consecutive days without any other intervention. (2) Model group: mice were fed a liquid diet with low alcohol concentration for the first 6 days as a transition and then changed to the Lieber–DeCarli alcoholic liquid diet with 5% alcohol concentration from the 7 th day for 28 days without drug intervention. (3) Germacrone group: The feed was treated the same as the model group, and Germacrone (molecular formula: C15H22O, purity > 98%, Yuanye, Shanghai, China, 10 mg/kg) [ , ] was given daily intragastric administration from day 8 for 21 days. (4) Brusatol group: the feed treatment was the same as the model group, and the Nrf2 antagonist Brusatol (Purity: 99.96%, Selleck, S7956, Shanghai, China, 2 mg/kg) [ ] was intraperitoneally injected every other day from the 8 th day for 21 days.

Techniques: Translocation Assay, Targeted Gene Expression, Western Blot, Expressing, Control, Reverse Transcription Polymerase Chain Reaction, Immunofluorescence

Journal: Chinese Medicine

Article Title: The effect and mechanism of Germacrone in ameliorating alcoholic fatty liver by inhibiting Nrf2/Rbp4

doi: 10.1186/s13020-025-01132-y

Figure Lengend Snippet: Germacrone inhibits Nrf2-dependent Rbp4 expression in the ALD model. A , B The protein expression levels of Rbp4 in the ALD model treated with Germacrone and Brusatol (n = 3). C Correlation analysis of liver TG content and Rbp4 expression. D Rbp4 mRNA expression in HepG2 cells in the Control group, Model group, Brusatol group, and Germacrone group was analyzed by RT-qPCR (n = 6). E , F Western blot was used to detect the expression level of Rbp4 protein under different conditions (n = 3). Compared with the model group, *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: After 3 days of adaptive feeding, C57BL/6 male mice were randomly divided into the following 4 groups with 6 mice in each group: (1) Control group: Mice were fed the Lieber–DeCarli standard liquid diet for 35 consecutive days without any other intervention. (2) Model group: mice were fed a liquid diet with low alcohol concentration for the first 6 days as a transition and then changed to the Lieber–DeCarli alcoholic liquid diet with 5% alcohol concentration from the 7 th day for 28 days without drug intervention. (3) Germacrone group: The feed was treated the same as the model group, and Germacrone (molecular formula: C15H22O, purity > 98%, Yuanye, Shanghai, China, 10 mg/kg) [ , ] was given daily intragastric administration from day 8 for 21 days. (4) Brusatol group: the feed treatment was the same as the model group, and the Nrf2 antagonist Brusatol (Purity: 99.96%, Selleck, S7956, Shanghai, China, 2 mg/kg) [ ] was intraperitoneally injected every other day from the 8 th day for 21 days.

Techniques: Expressing, Control, Quantitative RT-PCR, Western Blot

Journal: Chinese Medicine

Article Title: The effect and mechanism of Germacrone in ameliorating alcoholic fatty liver by inhibiting Nrf2/Rbp4

doi: 10.1186/s13020-025-01132-y

Figure Lengend Snippet: Mechanism of Germacrone in the Treatment of Alcohol-Induced Liver Disease (ALD). Under alcohol exposure, reactive oxygen species (ROS) and foreign electrophilic reagents inhibit the ubiquitination of Nrf2, leading to its nuclear translocation. In the nucleus, Nrf2 binds to antioxidant response elements (AREs), promoting the transcription of downstream target genes, including Rbp4 (lipid transport protein), HO-1, and Gsta1 (oxidative stress-regulated proteins). This process accelerates lipid transport, lipid accumulation, and oxidative damage in hepatocytes, which contributes to the progression of ALD (left panel). In contrast, Germacrone (Germacrone), the active component of Jia-Ga-Song-Tang (JGST), effectively inhibits Nrf2 activation and nuclear translocation, thereby down-regulating the expression of Rbp4, HO-1, and Gsta1. This reduces lipid accumulation and oxidative stress, protecting hepatocytes and improving liver health (right panel)

Article Snippet: After 3 days of adaptive feeding, C57BL/6 male mice were randomly divided into the following 4 groups with 6 mice in each group: (1) Control group: Mice were fed the Lieber–DeCarli standard liquid diet for 35 consecutive days without any other intervention. (2) Model group: mice were fed a liquid diet with low alcohol concentration for the first 6 days as a transition and then changed to the Lieber–DeCarli alcoholic liquid diet with 5% alcohol concentration from the 7 th day for 28 days without drug intervention. (3) Germacrone group: The feed was treated the same as the model group, and Germacrone (molecular formula: C15H22O, purity > 98%, Yuanye, Shanghai, China, 10 mg/kg) [ , ] was given daily intragastric administration from day 8 for 21 days. (4) Brusatol group: the feed treatment was the same as the model group, and the Nrf2 antagonist Brusatol (Purity: 99.96%, Selleck, S7956, Shanghai, China, 2 mg/kg) [ ] was intraperitoneally injected every other day from the 8 th day for 21 days.

Techniques: Ubiquitin Proteomics, Translocation Assay, Activation Assay, Expressing