Journal of Traditional Chinese Medicine ›› 2023, Vol. 43 ›› Issue (6): 1092-1102.DOI: 10.19852/j.cnki.jtcm.2023.06.004
• Research Articles • Previous Articles Next Articles
YIN Yixiao1,2,3, TANG Hao1,2,3, FANG Yi1,2,3, LIU Wei1,2,3, WANG Jun1,2,3, HU Yiyang4,5,6, PENG Jinghua1,2,3()
Received:
2022-09-11
Accepted:
2022-12-25
Online:
2023-10-25
Published:
2023-11-01
Contact:
PENG Jinghua, Institute of Liver diseases, Shuguang Hospital affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China; Key Laboratory of Liver and Kidney Diseases (Shanghai University of Traditional Chinese Medicine), Ministry of Education, Shanghai 201203, China; Shanghai Key Laboratory of Traditional Chinese Clinical Medicine, Shanghai 201203, China. pengjinghua2004@163.com. Telephone: +86-21-20256526
Supported by:
YIN Yixiao, TANG Hao, FANG Yi, LIU Wei, WANG Jun, HU Yiyang, PENG Jinghua. Hepatic transcriptome delineates the therapeutic effects of Sanren Tang (三仁汤) on high-fat diet-induced non-alcoholic fatty liver disease[J]. Journal of Traditional Chinese Medicine, 2023, 43(6): 1092-1102.
Group | n | Body weight (g) | Food intake (g/mouse per day) | ||||
---|---|---|---|---|---|---|---|
0 week | 4th week | 8th week | 12th week | 16th week | |||
Control | 10 | 22.94±0.71 | 24.94±1.67 | 27.22±2.22 | 29.04±2.59 | 30.10±2.97 | 2.18±0.12 |
HFD | 10 | 23.02±0.66 | 27.65±1.99a | 32.90±3.39a | 36.91±4.56a | 41.25±4.08a | 2.07±0.14 |
L-SRT | 10 | 22.90±0.64 | 27.69±1.71 | 33.27±2.98 | 36.69±4.85 | 36.39±5.08 | 2.03±0.11b |
H-SRT | 10 | 23.11±0.64 | 27.93±2.19 | 31.63±3.07 | 34.71±4.08 | 36.31±3.70 | 2.00±0.18a |
OCA | 10 | 23.11±0.57 | 27.99±1.28 | 33.03±2.09 | 36.69±4.27 | 37.80±4.22 | 1.96±0.14a |
Table 1 Body weight and food intake
Group | n | Body weight (g) | Food intake (g/mouse per day) | ||||
---|---|---|---|---|---|---|---|
0 week | 4th week | 8th week | 12th week | 16th week | |||
Control | 10 | 22.94±0.71 | 24.94±1.67 | 27.22±2.22 | 29.04±2.59 | 30.10±2.97 | 2.18±0.12 |
HFD | 10 | 23.02±0.66 | 27.65±1.99a | 32.90±3.39a | 36.91±4.56a | 41.25±4.08a | 2.07±0.14 |
L-SRT | 10 | 22.90±0.64 | 27.69±1.71 | 33.27±2.98 | 36.69±4.85 | 36.39±5.08 | 2.03±0.11b |
H-SRT | 10 | 23.11±0.64 | 27.93±2.19 | 31.63±3.07 | 34.71±4.08 | 36.31±3.70 | 2.00±0.18a |
OCA | 10 | 23.11±0.57 | 27.99±1.28 | 33.03±2.09 | 36.69±4.27 | 37.80±4.22 | 1.96±0.14a |
Group | n | TNF-α | IL-1β | IL-6 |
---|---|---|---|---|
Control | 10 | 1.00±0.27 | 1.00±0.13 | 1.00±0.25 |
HFD | 10 | 1.78±0.53a | 1.41±0.04a | 1.32±0.29a |
L-SRT | 10 | 1.19±0.56b | 0.84±0.27c | 0.93±0.38b |
H-SRT | 10 | 1.33±0.35b | 1.38±0.03 | 1.43±0.38 |
OCA | 10 | 1.67±0.41 | 0.79±0.35c | 0.63±0.17c |
Table 3 Relative mRNA expression of inflammatory factors in the liver tissue
Group | n | TNF-α | IL-1β | IL-6 |
---|---|---|---|---|
Control | 10 | 1.00±0.27 | 1.00±0.13 | 1.00±0.25 |
HFD | 10 | 1.78±0.53a | 1.41±0.04a | 1.32±0.29a |
L-SRT | 10 | 1.19±0.56b | 0.84±0.27c | 0.93±0.38b |
H-SRT | 10 | 1.33±0.35b | 1.38±0.03 | 1.43±0.38 |
OCA | 10 | 1.67±0.41 | 0.79±0.35c | 0.63±0.17c |
Figure 1 Sanren Tang ameliorates hepatic histology in NAFLD mice A: haematoxylin-eosin staining of the liver sections (× 200magnification). B: oil red staining of the liver sections (200 × magnification). 1: control group, 2: high-fat diet (HFD) group, 3: low-dose Sanren Tang (L-SRT) group, 4: high-dose Sanren Tang (H-SRT) group, 5: obeticholic acid (OCA) group. Mice in the control group were fed a control diet (D12450B, 10% kcal from fat). The others were fed HFD (D12492, 60% kcal from fat) for 16 weeks. From the 13th to 16th week, the mice in the L-SRT, H-SRT and OCA groups were administered with L-SRT (crude drug 1.09 g/mL, 20 mL/kg body weight), H-SRT (crude drug 2.18 g/mL, 20 mL/kg body weight), and OCA (10 mg/kg), respectively. The others were administered with an equal volume of double-distilled water.
Group | n | Fasting blood glucose (mg/dL) | Fasting insulin (ng/mL) | HOMA-IR | ALT (U/L) | Hepatic TG (mg/g tissue) | NAS |
---|---|---|---|---|---|---|---|
Control | 10 | 15.33±11.63 | 0.39±0.09 | 0.33±0.29 | 17.02±5.44 | 41.85±7.74 | 0 (0) |
HFD | 10 | 52.96±17.29a | 1.36±0.47a | 4.01±2.26a | 66.99±42.17a | 61.22±13.17a | 7 (2)a |
L-SRT | 10 | 56.15±13.16 | 0.69±0.25b | 2.03±0.87c | 24.93±26.46c | 40.17±16.27b | 1 (1.75)c |
H-SRT | 10 | 57.38±10.14 | 0.62±0.17b | 1.84±0.47d | 9.95±7.19c | 35.51±15.73b | 1 (1.5)c |
OCA | 10 | 49.69±19.34 | 1.23±0.64 | 2.69±1.58 | 24.69±19.01 | 41.81±10.88b | 1 (1)b |
Table 2 Laboratory and pathological parameters of the mice
Group | n | Fasting blood glucose (mg/dL) | Fasting insulin (ng/mL) | HOMA-IR | ALT (U/L) | Hepatic TG (mg/g tissue) | NAS |
---|---|---|---|---|---|---|---|
Control | 10 | 15.33±11.63 | 0.39±0.09 | 0.33±0.29 | 17.02±5.44 | 41.85±7.74 | 0 (0) |
HFD | 10 | 52.96±17.29a | 1.36±0.47a | 4.01±2.26a | 66.99±42.17a | 61.22±13.17a | 7 (2)a |
L-SRT | 10 | 56.15±13.16 | 0.69±0.25b | 2.03±0.87c | 24.93±26.46c | 40.17±16.27b | 1 (1.75)c |
H-SRT | 10 | 57.38±10.14 | 0.62±0.17b | 1.84±0.47d | 9.95±7.19c | 35.51±15.73b | 1 (1.5)c |
OCA | 10 | 49.69±19.34 | 1.23±0.64 | 2.69±1.58 | 24.69±19.01 | 41.81±10.88b | 1 (1)b |
Figure 2 Hepatic transcriptome profile and hepatic genes reversed by SRT in HFD-induced NAFLD A: PCA of transcriptome data; B: clustering analysis of sample-to-sample distances; C: number of DEGs; D: DEG intersection; E: heatmap of the top 20 genes up-or down-regulated by HFD and reversed by H-SRT; F: GO analysis of total genes regulated by HFD and reversed by H-SRT; G: KEGG pathway enrichment analysis of total genes regulated by HFD and reversed by H-SRT. PCA: principal component analysis; DEGs: differentially expressed genes; ECM: extracellular matrix; HFD: high-fat diet; NAFLD: non-alcoholic fatty liver disease; H-SRT: high-dose of Sanren Tang; GO: gene ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes. Samples from the control, HFD, and H-SRT groups (4 samples per group) were randomly chosen for RNA-seq. Mice in the control group were fed a control diet (D12450B, 10% kcal from fat). The others were fed HFD (D12492, 60% kcal from fat) for 16 weeks. From the 13th to 16th week, the mice in the L-SRT, H-SRT and OCA groups were administered with L-SRT (crude drug 1.09 g/mL, 20 mL/kg body weight), H-SRT (crude drug 2.18 g/mL, 20 mL/kg body weight), and OCA (10 mg/kg), respectively. The others were administered with an equal volume of double-distilled water.
Figure 3 Verification of the genes enriched in the top three KEGG pathways A: relative hepatic mRNA expression levels of genes enriched in the top three KEGG pathways; B: representative images of Western-blotting of PPARγ and IRF 6 in liver tissue; C: the densities analysis of Western-blotting and of PPARγ; D: the densities analysis of Western-blotting band of IRF6. 1: control; 2: HFD; 3: H-SRT; 4: L-SRT; 5: OCA. KEGG: Kyoto Encyclopedia of Genes and Genomes; PPARγ: proliferator-activated receptor γ; IRF: interferon regulatory factor; HFD: high-fat diet; L-SRT: low-dose Sanren Tang; H-SRT: high-dose Sanren Tang; OCA: obeticholic acid; GAPDH: glyceraldehyde 3-phosphate dehydrogenase. Ten samples per group were used for detection of mRNA expression. The data of Western-blotting were obtained from three samples per group chosen randomly. aP < 0.01, vs control, bP < 0.01, vs HFD, cP < 0.05, vs control, dP < 0.05, vs HFD. The original uncut image of the Western-blot can be found in supplementary Figures7 and 8. Mice in the control group were fed a control diet (D12450B, 10% kcal from fat). The others were fed HFD (D12492, 60% kcal from fat) for 16 weeks. From the 13th to 16th week, the mice in the L-SRT, H-SRT and OCA groups were administered with L-SRT (crude drug 1.09 g/mL, 20 mL/kg body weight), H-SRT (crude drug 2.18 g/mL, 20 mL/kg body weight), and OCA (10 mg/kg), respectively. The others were administered with an equal volume of double-distilled water.
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