Journal of Traditional Chinese Medicine ›› 2026, Vol. 46 ›› Issue (4): 873-882.DOI: 10.19852/j.cnki.jtcm.2026.04.008
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Taohong Huazhuo decoction (桃红化浊汤) alleviates carbon tetrachloride-induced hepatic fibrosis in rats by inhibiting the nicotinamide adenine dinucleotide phosphate oxidase 4/nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 signaling pathway
REN Yuan1, CHEN Xiangni1, SHI Lei1, MAO Minghua1, LI Shufang1, YAO Jia2, HAO Jianmei1(
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Department of Hepatology ,Xi'an Hospital of Traditional Chinese Medicine Xi'an 710021, China
2Experimental Center ,Xi'an Hospital of Traditional Chinese Medicine Xi'an 710021, China
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Received:2025-06-05Accepted:2025-11-04Online:2026-08-15Published:2026-08-08 -
Contact:Prof. HAO Jianmei, Department of Hepatology, Xi'an Hospital of Traditional Chinese Medicine, Xi'an 710021, China. hjmyzp@163.com,Telephone: +86-15129629909
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Supported by:Youth Program of Xi'an Municipal Health Commission of China: Exploring the Molecular Mechanism of Taohong Huazhuo decoction in Improving Liver Fibrosis by Regulating the Nicotinamide Adenine Dinucleotide Phosphate Oxidase 2/Reactive Oxygen Species Signaling Pathway Based on the Theory of Damp-heat and Ministerial Fire(2024qn20);Cultivation Projects of the National Natural Science Foundation of Xi'an Traditional Chinese Medicine Hospital: Exploring the Molecular Mechanism of Taohong Huazhuo decoction Against Liver Fibrosis Based on Endoplasmic Reticulum Stress-mediated Ferroptosis(2024YJ02)
Cite this article
REN Yuan, CHEN Xiangni, SHI Lei, MAO Minghua, LI Shufang, YAO Jia, HAO Jianmei. Taohong Huazhuo decoction (桃红化浊汤) alleviates carbon tetrachloride-induced hepatic fibrosis in rats by inhibiting the nicotinamide adenine dinucleotide phosphate oxidase 4/nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 signaling pathway[J]. Journal of Traditional Chinese Medicine, 2026, 46(4): 873-882.
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Table 1 THHZD improved liver function in rats with CCl4-induced hepatic fibrosis ($\bar{x}±s$)
| Group | n | ALT (U/L) | AST (U/L) | TBIL (μmol/L) |
|---|---|---|---|---|
| Normal | 8 | 52.23±7.17 | 156.00±28.65 | 0.70±0.14a |
| Model | 8 | 437.35±59.65a | 538.50±99.67a | 6.99±1.21 |
| Colchicine | 8 | 279.70±112.33b | 319.75±107.62c | 4.49±1.49b |
| THHZD-L | 8 | 309.38±110.19 | 371.00±98.33b | 5.01±1.50 |
| THHZD-M | 8 | 279.65±34.77b | 296.50±56.20c | 4.47±0.88b |
| THHZD-H | 8 | 233.05±49.9c | 307.50±86.34c | 3.45±1.75c |
Table 1 THHZD improved liver function in rats with CCl4-induced hepatic fibrosis ($\bar{x}±s$)
| Group | n | ALT (U/L) | AST (U/L) | TBIL (μmol/L) |
|---|---|---|---|---|
| Normal | 8 | 52.23±7.17 | 156.00±28.65 | 0.70±0.14a |
| Model | 8 | 437.35±59.65a | 538.50±99.67a | 6.99±1.21 |
| Colchicine | 8 | 279.70±112.33b | 319.75±107.62c | 4.49±1.49b |
| THHZD-L | 8 | 309.38±110.19 | 371.00±98.33b | 5.01±1.50 |
| THHZD-M | 8 | 279.65±34.77b | 296.50±56.20c | 4.47±0.88b |
| THHZD-H | 8 | 233.05±49.9c | 307.50±86.34c | 3.45±1.75c |
Figure 1 THHZD inhibited CCL4-induced hepatic fibrosis and hepatic stellate cell activation in rats A: hematoxylin and eosin staining of hepatic tissues (× 100, scale bars = 100 μm); B: Western blot detection of collagen I protein expression in hepatic tissues; C: densitometric quantification of collagen I in hepatic tissues; D: masson's trichrome staining of hepatic tissues (× 100, scale bars = 100 μm); E: interstitial collagen was determined using Masson's trichrome staining; F: images of immunohistochemistry staining of α-SMA (× 200, scale bars = 40 μm); G: statistical analysis of immunohistochemistry staining of α-SMA; A1, D1, F1: normal group; A2, D2, F2: model group; A3, D3, F3: colchicine group; A4, D4, F4: THHZD-L group; A5, D5, F5: THHZD-M group; A6, D6, F6: THHZD-H group. Normal group: daily saline gavage for 4 weeks; Model group: daily saline gavage for 4 weeks; Colchicine group: daily intragastric colchicine (0.1 mg/kg) for 4 weeks; THHZD-L group: daily intragastric THHZD (3.285 g/kg) for 4 weeks; THHZD-M group: daily intragastric THHZD (6.57 g/kg) for 4 weeks; THHZD-H group: daily intragastric THHZD (13.14 g/kg) for 4 weeks. α-SMA: alpha-smooth muscle actin; THHZD-L: low-dose of Taohong Huazhuo decoction; THHZD-M: medium-dose of Taohong Huazhuo decoction; THHZD-H: high-dose of Taohong Huazhuo decoction. Statistical comparisons among groups were conducted via one-way analysis of variance, followed by Tukey’s test for pairwise multiple comparisons. All data are expressed as mean ± standard deviation (n = 3). Compared to the normal group, aP < 0.01; compared to the model group, bP < 0.01 and cP < 0.05.
Figure 1 THHZD inhibited CCL4-induced hepatic fibrosis and hepatic stellate cell activation in rats A: hematoxylin and eosin staining of hepatic tissues (× 100, scale bars = 100 μm); B: Western blot detection of collagen I protein expression in hepatic tissues; C: densitometric quantification of collagen I in hepatic tissues; D: masson's trichrome staining of hepatic tissues (× 100, scale bars = 100 μm); E: interstitial collagen was determined using Masson's trichrome staining; F: images of immunohistochemistry staining of α-SMA (× 200, scale bars = 40 μm); G: statistical analysis of immunohistochemistry staining of α-SMA; A1, D1, F1: normal group; A2, D2, F2: model group; A3, D3, F3: colchicine group; A4, D4, F4: THHZD-L group; A5, D5, F5: THHZD-M group; A6, D6, F6: THHZD-H group. Normal group: daily saline gavage for 4 weeks; Model group: daily saline gavage for 4 weeks; Colchicine group: daily intragastric colchicine (0.1 mg/kg) for 4 weeks; THHZD-L group: daily intragastric THHZD (3.285 g/kg) for 4 weeks; THHZD-M group: daily intragastric THHZD (6.57 g/kg) for 4 weeks; THHZD-H group: daily intragastric THHZD (13.14 g/kg) for 4 weeks. α-SMA: alpha-smooth muscle actin; THHZD-L: low-dose of Taohong Huazhuo decoction; THHZD-M: medium-dose of Taohong Huazhuo decoction; THHZD-H: high-dose of Taohong Huazhuo decoction. Statistical comparisons among groups were conducted via one-way analysis of variance, followed by Tukey’s test for pairwise multiple comparisons. All data are expressed as mean ± standard deviation (n = 3). Compared to the normal group, aP < 0.01; compared to the model group, bP < 0.01 and cP < 0.05.
Figure 2 THHZD inhibits NOX4-mediated oxidative stress in the livers of rats with CCl4-induced hepatic fibrosis A: ROS expression in hepatic tissues was detected by immunofluorescence staining (× 200, scale bars = 50 μm). A1, A7, A13: normal group; A2, A8, A14: model group; A3, A9, A15: colchicine group; A4, A10, A16: THHZD-L group; A5, A11, A17: THHZD-M group; A6, A12, A18: THHZD-H group; B: statistical analysis of immunofluorescence of ROS; C: western blot detection of NOX4 protein expression in hepatic tissues; D: densitometric quantification of NOX4 in hepatic tissues; E: the levels of MDA in hepatic tissues were determined using ELISA kits; F: the levels of GSH in hepatic tissues were determined using ELISA kits. Normal group: daily saline gavage for 4 weeks; Model group: daily saline gavage for 4 weeks; Colchicine group: daily intragastric colchicine (0.1 mg/kg) for 4 weeks; THHZD-L group: daily intragastric THHZD (3.285 g/kg) for 4 weeks; THHZD-M group: daily intragastric THHZD (6.57 g/kg) for 4 weeks; THHZD-H group: daily intragastric THHZD (13.14 g/kg) for 4 weeks. GSH: glutathione; MDA: malondialdehyde; IL-1β: interleukin-1β; IL-18: interleukin-18; ROS: reactive oxygen species; NOX4: nicotinamide adenine dinucleotide phosphate oxidase 4; THHZD-L: low-dose of Taohong Huazhuo decoction; THHZD-M: medium-dose of Taohong Huazhuo decoction; THHZD-H: high-dose of Taohong Huazhuo decoction; ELISA: enzyme-linked immunosorbent assay. Statistical comparisons among groups were conducted via one-way analysis of variance, followed by Tukey’s test for pairwise multiple comparisons. All data are expressed as mean ± standard deviation (n = 3). Compared to the normal group, aP < 0.01; compared to the model group, bP < 0.01 and cP< 0.05.
Figure 2 THHZD inhibits NOX4-mediated oxidative stress in the livers of rats with CCl4-induced hepatic fibrosis A: ROS expression in hepatic tissues was detected by immunofluorescence staining (× 200, scale bars = 50 μm). A1, A7, A13: normal group; A2, A8, A14: model group; A3, A9, A15: colchicine group; A4, A10, A16: THHZD-L group; A5, A11, A17: THHZD-M group; A6, A12, A18: THHZD-H group; B: statistical analysis of immunofluorescence of ROS; C: western blot detection of NOX4 protein expression in hepatic tissues; D: densitometric quantification of NOX4 in hepatic tissues; E: the levels of MDA in hepatic tissues were determined using ELISA kits; F: the levels of GSH in hepatic tissues were determined using ELISA kits. Normal group: daily saline gavage for 4 weeks; Model group: daily saline gavage for 4 weeks; Colchicine group: daily intragastric colchicine (0.1 mg/kg) for 4 weeks; THHZD-L group: daily intragastric THHZD (3.285 g/kg) for 4 weeks; THHZD-M group: daily intragastric THHZD (6.57 g/kg) for 4 weeks; THHZD-H group: daily intragastric THHZD (13.14 g/kg) for 4 weeks. GSH: glutathione; MDA: malondialdehyde; IL-1β: interleukin-1β; IL-18: interleukin-18; ROS: reactive oxygen species; NOX4: nicotinamide adenine dinucleotide phosphate oxidase 4; THHZD-L: low-dose of Taohong Huazhuo decoction; THHZD-M: medium-dose of Taohong Huazhuo decoction; THHZD-H: high-dose of Taohong Huazhuo decoction; ELISA: enzyme-linked immunosorbent assay. Statistical comparisons among groups were conducted via one-way analysis of variance, followed by Tukey’s test for pairwise multiple comparisons. All data are expressed as mean ± standard deviation (n = 3). Compared to the normal group, aP < 0.01; compared to the model group, bP < 0.01 and cP< 0.05.
Figure 3 THHZD inhibits hepatic inflammasome activation in rats with CCl4-induced hepatic fibrosis A: Western blot detection of NLRP3, caspase-1, IL-1β, and IL-18 protein expression in hepatic tissues; B: densitometric quantification of NLRP3 in hepatic tissues; C: densitometric quantification of caspase-1 in hepatic tissues; D: densitometric quantification of IL-1β in hepatic tissues; E: densitometric quantification of IL-8 in hepatic tissues; F: the levels of serum IL-1β were determined using ELISA kits; G: the levels of serum IL-18 were determined using ELISA kits; Normal group: daily saline gavage for 4 weeks; Model group: daily saline gavage for 4 weeks; Colchicine group: daily intragastric colchicine (0.1 mg/kg) for 4 weeks; THHZD-L group: daily intragastric THHZD (3.285 g/kg) for 4 weeks; THHZD-M group: daily intragastric THHZD (6.57 g/kg) for 4 weeks; THHZD-H group: daily intragastric THHZD (13.14 g/kg) for 4 weeks. IL-1β: interleukin-1β; IL-18: interleukin-18; NLRP3: nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3; THHZD-L: low-dose of Taohong Huazhuo decoction; THHZD-M: medium-dose of Taohong Huazhuo decoction; THHZD-H: high-dose of Taohong Huazhuo decoction; ELISA: enzyme-linked immunosorbent assay. Statistical comparisons among groups were conducted via one-way analysis of variance, followed by Tukey’s test for pairwise multiple comparisons. All data are expressed as mean ± standard deviation (n = 3). Compared to the normal group, aP < 0.01; compared to the model group, bP < 0.05 and cP< 0.01.
Figure 3 THHZD inhibits hepatic inflammasome activation in rats with CCl4-induced hepatic fibrosis A: Western blot detection of NLRP3, caspase-1, IL-1β, and IL-18 protein expression in hepatic tissues; B: densitometric quantification of NLRP3 in hepatic tissues; C: densitometric quantification of caspase-1 in hepatic tissues; D: densitometric quantification of IL-1β in hepatic tissues; E: densitometric quantification of IL-8 in hepatic tissues; F: the levels of serum IL-1β were determined using ELISA kits; G: the levels of serum IL-18 were determined using ELISA kits; Normal group: daily saline gavage for 4 weeks; Model group: daily saline gavage for 4 weeks; Colchicine group: daily intragastric colchicine (0.1 mg/kg) for 4 weeks; THHZD-L group: daily intragastric THHZD (3.285 g/kg) for 4 weeks; THHZD-M group: daily intragastric THHZD (6.57 g/kg) for 4 weeks; THHZD-H group: daily intragastric THHZD (13.14 g/kg) for 4 weeks. IL-1β: interleukin-1β; IL-18: interleukin-18; NLRP3: nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3; THHZD-L: low-dose of Taohong Huazhuo decoction; THHZD-M: medium-dose of Taohong Huazhuo decoction; THHZD-H: high-dose of Taohong Huazhuo decoction; ELISA: enzyme-linked immunosorbent assay. Statistical comparisons among groups were conducted via one-way analysis of variance, followed by Tukey’s test for pairwise multiple comparisons. All data are expressed as mean ± standard deviation (n = 3). Compared to the normal group, aP < 0.01; compared to the model group, bP < 0.05 and cP< 0.01.
Figure 4 THHZD effectively suppresses the activation of HSCs via blocking the activation of the NOX4/NLRP3 signaling pathway in vitro A: effect of THHZD on the viability of HSC-T6 cells was evaluated using CCK-8 assay; B: Western blot detection of α-SMA, collagen I, NOX4, NLRP3, caspase-1, IL-1β, and IL-18 protein expression in HSC-T6 cells; C: densitometric quantification of α-SMA; D: densitometric quantification of collagen I; E: densitometric quantification of NOX4; F: densitometric quantification of NLRP3; G: densitometric quantification of caspase-1; H: densitometric quantification of IL-1β; I: densitometric quantification of IL-18; Normal group: no treatment; Model group: 10 ng/mL TGF-β1, 24 h; THHZD-L: 10 ng/mL TGF-β1 + 0.15 mg/mL THHZD, 24 h; THHZD-M: 10 ng/mL TGF-β1 + 0.3 mg/mL THHZD, 24 h; THHZD-H: 10 ng/mL TGF-β1 + 0.6 mg/mL THHZD, 24 h. IL-1β: interleukin-1β; IL-18: interleukin-18; NLRP3: nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3; NOX4: nicotinamide adenine dinucleotide phosphate oxidase 4; α-SMA: alpha-smooth muscle actin; HSCs: hepatic stellate cells; TGF-β1: transforming growth factor-β1; CCK-8: cell counting kit-8; THHZD-L: low-dose of Taohong Huazhuo decoction; THHZD-M: medium-dose of Taohong Huazhuo decoction; THHZD-H: high-dose of Taohong Huazhuo decoction. Statistical comparisons among groups were conducted via one-way analysis of variance, followed by Tukey’s test for pairwise multiple comparisons. All data are expressed as mean ± standard deviation (n = 3). Compared to the normal group, aP < 0.01; compared to the model group, bP < 0.05 and cP < 0.01.
Figure 4 THHZD effectively suppresses the activation of HSCs via blocking the activation of the NOX4/NLRP3 signaling pathway in vitro A: effect of THHZD on the viability of HSC-T6 cells was evaluated using CCK-8 assay; B: Western blot detection of α-SMA, collagen I, NOX4, NLRP3, caspase-1, IL-1β, and IL-18 protein expression in HSC-T6 cells; C: densitometric quantification of α-SMA; D: densitometric quantification of collagen I; E: densitometric quantification of NOX4; F: densitometric quantification of NLRP3; G: densitometric quantification of caspase-1; H: densitometric quantification of IL-1β; I: densitometric quantification of IL-18; Normal group: no treatment; Model group: 10 ng/mL TGF-β1, 24 h; THHZD-L: 10 ng/mL TGF-β1 + 0.15 mg/mL THHZD, 24 h; THHZD-M: 10 ng/mL TGF-β1 + 0.3 mg/mL THHZD, 24 h; THHZD-H: 10 ng/mL TGF-β1 + 0.6 mg/mL THHZD, 24 h. IL-1β: interleukin-1β; IL-18: interleukin-18; NLRP3: nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3; NOX4: nicotinamide adenine dinucleotide phosphate oxidase 4; α-SMA: alpha-smooth muscle actin; HSCs: hepatic stellate cells; TGF-β1: transforming growth factor-β1; CCK-8: cell counting kit-8; THHZD-L: low-dose of Taohong Huazhuo decoction; THHZD-M: medium-dose of Taohong Huazhuo decoction; THHZD-H: high-dose of Taohong Huazhuo decoction. Statistical comparisons among groups were conducted via one-way analysis of variance, followed by Tukey’s test for pairwise multiple comparisons. All data are expressed as mean ± standard deviation (n = 3). Compared to the normal group, aP < 0.01; compared to the model group, bP < 0.05 and cP < 0.01.
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