Journal of Traditional Chinese Medicine ›› 2024, Vol. 44 ›› Issue (4): 642-651.DOI: 10.19852/j.cnki.jtcm.2024.04.006
• Original articles • Previous Articles Next Articles
LYU Meixian1, ZHOU Huan2, ZHI Limin3, ZHOU Jinling4, GAN Rizhi5, QIN Yanping6, HE Nengting7, ZUO Qiqi8, LI Hao1, DONG Min1, LIANG Gang1()
Received:
2023-03-21
Accepted:
2023-08-29
Online:
2024-08-15
Published:
2024-07-15
Contact:
LIANG Gang, Pharmaceutical College, Guangxi Medical University, Nanning 530022, China. Supported by:
LYU Meixian, ZHOU Huan, ZHI Limin, ZHOU Jinling, GAN Rizhi, QIN Yanping, HE Nengting, ZUO Qiqi, LI Hao, DONG Min, LIANG Gang. Saponin Ⅰ from Shuitianqi (Rhizoma Schizocapasae Plantagineae) inhibits metastasis by negatively regulating the transforming growth factor-β1/Smad7 network and epithelial-mesenchymal transition in the intrahepatic metastasis Bagg’s Albino/c mouse model[J]. Journal of Traditional Chinese Medicine, 2024, 44(4): 642-651.
Figure 1 Effect of SSPH Ⅰ on intrahepatic metastasis and TGF-β1/Smad7 signaling pathway A: hematoxylin-eosin staining was used to analyze intrahepatic metastasis in model group, cisplatin group and SSPH Ⅰ groups. A1: model group; A2: cisplatin group; A3: SSPH Ⅰ (25 mg/kg) group; A4: SSPH Ⅰ (50 mg/kg) group; A5: SSPH Ⅰ (75 mg/kg) group. B: Western blotting was used to detect the expression of TGF-β1 and Smad7 proteins. 1: model group; 2: cisplatin group; 3: SSPH Ⅰ (25 mg/kg) group; 4: SSPH Ⅰ (50 mg/kg) group; 5: SSPH Ⅰ (75 mg/kg) group. C: quantitative analysis of TGF-β1 and Smad7 proteins expression. D: immunohistochemical assay was used to detect the expression of TGF-β1 protein in model group, cisplatin group and SSPH Ⅰ groups. D1: model group; D2: cisplatin group; D3: SSPH Ⅰ (25 mg/kg) group; D4: SSPH Ⅰ (50 mg/kg) group; D5: SSPH Ⅰ (75 mg/kg) group. magnification at ×200; scale bar, 20 μm, dyeing method of all pictures are the immunohistochemical DAB method. E: immunohistochemical assay was used to detect the expression of Smad7 protein in model group, cisplatin group and SSPH Ⅰ groups. E1: model group; E2: cisplatin group; E3: SSPH Ⅰ (25 mg/kg) group; E4: SSPH Ⅰ (50 mg/kg) group; E5: SSPH Ⅰ (75 mg/kg) group. magnification at ×200; scale bar, 20 μm, dyeing method of all pictures are the immunohistochemical DAB method. F: quantitative analysis of TGF-β1 and Smad7 proteins expression. G: enzyme linked immunosorbent assay was used to detect the expression of TGF-β1 protein in serum. Model group: free diet + gavage with equal amount of normal saline per day; cisplatin group: model group + intraperitoneal injection with equal amount of cisplatin 2 mg/kg every other day; SSPH Ⅰ (25 mg/kg) group: model group + gavage with equal amount of SSPH Ⅰ 25 mg/kg per day; SSPH Ⅰ (50 mg/kg) group: model group + gavage with equal amount of SSPH Ⅰ 50 mg/kg per day; SSPH Ⅰ (75 mg/kg) group: model group + gavage with equal amount of SSPH Ⅰ 75 mg/kg per day. SSPH Ⅰ: Saponin I from Shuitianqi (Rhizoma Schizocapasae Plantagineae); TGF-β1: transforming growth factor-β1; DAB: diaminobenzidine. All data was measured by one-way analysis, and Newman-Keuls test was performed for inter-group comparisons. All data was presented as mean ± standard deviation (n = 3). aP < 0.01, bP < 0.05 compared to the model group.
Figure 2 Effect of SSPH Ⅰ on EMT A: western blotting was used to detect the expression of N-cadherin, E-cadherin and Vimentin proteins. A1: model group; A2: cisplatin group; A3: SSPH Ⅰ (25 mg/kg) group; A4: SSPH Ⅰ (50 mg/kg) group; A5: SSPH Ⅰ (75 mg/kg) group. B: quantitative analysis of N-cadherin, E-cadherin and Vimentin proteins expression. C: immunohistochemical assay was used to detect the expression of E-cadherin protein in model group, cisplatin group and SSPH Ⅰ groups. C1: model group; C2: cisplatin group; C3: SSPH Ⅰ (25 mg/kg) group; C4: SSPH Ⅰ (50 mg/kg) group; C5: SSPH Ⅰ (75 mg/kg) group. magnification at ×200; scale bar, 20 μm, dyeing method of all pictures are the immunohistochemical DAB method. D: immunohistochemical assay was used to detect the expression of N-cadherin protein in model group, cisplatin group and SSPH Ⅰ groups. D1: model group; D2: cisplatin group; D3: SSPH Ⅰ (25 mg/kg) group; E4: SSPH Ⅰ (50 mg/kg) group; D5: SSPH Ⅰ (75 mg/kg) group. magnification at ×200; scale bar, 20 μm, dyeing method of all pictures are the immunohistochemical DAB method. E: immunohistochemical assay was used to detect the expression of Vimentin protein in model group, cisplatin group and SSPH Ⅰ groups. E1: model group; E2: cisplatin group; E3: SSPH Ⅰ (25 mg/kg) group; E4: SSPH Ⅰ (50 mg/kg) group; E5: SSPH Ⅰ (75 mg/kg) group. magnification at ×200; scale bar, 20 μm, dyeing method of all pictures are the immunohistochemical DAB method. F: quantitative analysis of N-cadherin, E-cadherin and Vimentin proteins expression. model group: free diet + gavage with equal amount of normal saline per day; cisplatin group: model group + intraperitoneal injection with equal amount of cisplatin 2 mg/kg every other day; SSPH Ⅰ (25 mg/kg) group: model group + gavage with equal amount of SSPH Ⅰ 25 mg/kg per day; SSPH Ⅰ (50 mg/kg) group: model group + gavage with equal amount of SSPH Ⅰ 50 mg/kg per day; SSPH Ⅰ (75 mg/kg) group: model group + gavage with equal amount of SSPH Ⅰ 75 mg/kg per day. SSPH Ⅰ: Saponin I from Shuitianqi (Rhizoma Schizocapasae Plantagineae); EMT: epithelial-mesenchymal transition; DAB: diaminobenzidine. All data was measured by one-way analysis, and Newman-Keuls test was performed for inter-group comparisons. All data was presented as mean ± standard deviation (n = 3). aP < 0.05, bP < 0.001 compared to the model group.
Figure 3 Effect of SSPH Ⅰ on MMPs A: Western blotting was used to detect the expression of MMP-2 and MMP-9 proteins. A1: model group; A2: cisplatin group; A3: SSPH Ⅰ (25 mg/kg) group; A4: SSPH Ⅰ (50 mg/kg) group; A5: SSPH Ⅰ (75 mg/kg) group. B: quantitative analysis of MMP-2 and MMP-9 proteins expression. C: immunohistochemical assay was used to detect the expression of MMP-2 protein in model group, cisplatin group and SSPH Ⅰ groups. C1: model group; C2: cisplatin group; C3: SSPH Ⅰ (25 mg/kg) group; C4: SSPH Ⅰ (50 mg/kg) group; C5: SSPH Ⅰ (75 mg/kg) group. magnification at ×200; scale bar, 20 μm, dyeing method of all pictures are the immunohistochemical DAB method. D: immunohistochemical assay was used to detect the expression of MMP-9 protein in model group, cisplatin group and SSPH Ⅰ groups. D1: model group; D2: cisplatin group; D3: SSPH Ⅰ (25 mg/kg) group; D4: SSPH Ⅰ (50 mg/kg) group; D5: SSPH Ⅰ (75 mg/kg) group. magnification at ×200; scale bar, 20 μm, dyeing method of all pictures are the immunohistochemical DAB method. E: quantitative analysis of MMP-2 and MMP-9 proteins expression. F: enzyme linked immunosorbent assay was used to detect the expression of MMP-2 protein in serum. G: enzyme linked immunosorbent assay was used to detect the expression of MMP-9 protein in serum. Model group: free diet + gavage with equal amount of normal saline per day; cisplatin group: model group + intraperitoneal injection with equal amount of cisplatin 2 mg/kg every other day; SSPH Ⅰ (25 mg/kg) group: model group + gavage with equal amount of SSPH Ⅰ 25 mg/kg per day; SSPH Ⅰ (50 mg/kg) group: model group + gavage with equal amount of SSPH Ⅰ 50 mg/kg per day; SSPH Ⅰ (75 mg/kg) group: model group + gavage with equal amount of SSPH Ⅰ 75 mg/kg per day. SSPH Ⅰ: Saponin I from Shuitianqi (Rhizoma Schizocapasae Plantagineae); MMP: matrix etalloproteinase; DAB: diaminobenzidine. All data was measured by one-way analysis, and Newman-Keuls test was performed for inter-group comparisons. All data was presented as mean ± standard deviation (n = 3). aP < 0.05, bP < 0.001 compared to the model group.
Figure 4 Effect of SSPH Ⅰ on angiogenesis A: Western blotting was used to detect the expression of CD31, CD34 and VEGF proteins. A1: model group; A2: cisplatin group; A3: SSPH Ⅰ (25 mg/kg) group; A4: SSPH Ⅰ (50 mg/kg) group; A5: SSPH Ⅰ (75 mg/kg) group. B: quantitative analysis of CD31, CD34 and VEGF proteins expression. C: immunohistochemical assay was used to detect the expression of CD31 protein in model group, cisplatin group and SSPH Ⅰ groups. C1: model group; C2: cisplatin group; C3: SSPH Ⅰ (25 mg/kg) group; C4: SSPH Ⅰ (50 mg/kg) group; C5: SSPH Ⅰ (75 mg/kg) group. magnification at ×200; scale bar, 20 μm, dyeing method of all pictures are the immunohistochemical DAB method. D: immunohistochemical assay was used to detect the expression of CD34 protein in model group, cisplatin group and SSPH Ⅰ groups. D1: model group; D2: cisplatin group; D3: SSPH Ⅰ (25 mg/kg) group; D4: SSPH Ⅰ (50 mg/kg) group; D5: SSPH Ⅰ (75 mg/kg) group. magnification at ×200; scale bar, 20 μm, dyeing method of all pictures are the immunohistochemical DAB method. E: immunohistochemical assay was used to detect the expression of VEGF protein in model group, cisplatin group and SSPH Ⅰ groups. E1: model group; E2: cisplatin group; E3: SSPH Ⅰ (25 mg/kg) group; E4: SSPH Ⅰ (50 mg/kg) group; E5: SSPH Ⅰ (75 mg/kg) group. magnification at ×200; scale bar, 20 μm, dyeing method of all pictures are the immunohistochemical DAB method. F: quantitative analysis of CD31, CD34 and VEGF proteins expression. G: enzyme linked immunosorbent assay was used to detect the expression of VEGF protein in serum. Model group: free diet + gavage with equal amount of normal saline per day; cisplatin group: model group + intraperitoneal injection with equal amount of cisplatin 2 mg/kg every other day; SSPH Ⅰ (25 mg/kg) group: model group + gavage with equal amount of SSPH Ⅰ 25 mg/kg per day; SSPH Ⅰ (50 mg/kg) group: model group + gavage with equal amount of SSPH Ⅰ 50 mg/kg per day; SSPH Ⅰ (75 mg/kg) group: model group + gavage with equal amount of SSPH Ⅰ 75 mg/kg per day. SSPH Ⅰ: Saponin I from Shuitianqi (Rhizoma Schizocapasae Plantagineae); VEGF: vascular endothelial growth factor; DAB: diaminobenzidine. All data was measured by one-way analysis, and Newman-Keuls test was performed for inter-group comparisons. All data was presented as mean ± standard deviation (n = 3). aP < 0.05, bP < 0.001 compared to the model group.
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