Journal of Traditional Chinese Medicine ›› 2024, Vol. 44 ›› Issue (4): 703-712.DOI: 10.19852/j.cnki.jtcm.20240617.002
• Original articles • Previous Articles Next Articles
GUO Yuxi1, LI Ze1, CHENG Nan1, JIA Xuemei1, WANG Jie1, MA Hongyu2, ZHAO Runyuan3, LI Bolin1, XUE Yucong4, CAI Yanru1(), YANG Qian1()
Received:
2023-03-22
Accepted:
2023-07-24
Online:
2024-08-15
Published:
2024-06-17
Contact:
YANG Qian, Department of spleen and stomach diseases, First Affiliated Hospital of Hebei University of Traditional Chinese Medicine, Shijiazhuang 050000, China. Supported by:
GUO Yuxi, LI Ze, CHENG Nan, JIA Xuemei, WANG Jie, MA Hongyu, ZHAO Runyuan, LI Bolin, XUE Yucong, CAI Yanru, YANG Qian. High-throughput sequencing analysis of differential microRNA expression in the process of blocking the progression of chronic atrophic gastritis to gastric cancer by Xianglian Huazhuo formula (香连化浊方)[J]. Journal of Traditional Chinese Medicine, 2024, 44(4): 703-712.
Figure 1 Histologic changes in the gastric mucosal tissue (magnification × 200, scale bar 50 μm) A: normal group; B: model group; C: XLHZ group. normal group: drink and eat freely + the same volume of normal saline was given by gavage; model group: MNNG, 180 μg/mL + 2% sodium salicylate, 1 mL/100 g per day + hunger and satiety disorders; XLHZ: model group + XLHZ, 36 g·kg-1·d-1. HE: hematoxylin and eosin; CAG: chronic atrophic gastritis; XLHZ: Xianglian Huazhuo formula.
Group | n | Normal | Model | XLHZ |
---|---|---|---|---|
miR-20a-3p | 3 | 1.08±0.49 | 6.65±1.38a | 3.31±0.76b |
miR-320-3p | 3 | 1.04±0.31 | 0.15±0.06a | 0.44±0.15b |
miR-34b-5p | 3 | 1.05±0.37 | 0.11±0.07a | 0.58±0.13c |
miR-483-3p | 3 | 1.01±0.15 | 0.29±0.16a | 0.37±0.12 |
miR-883-3p | 3 | 1.03±0.28 | 0.21±0.05a | 0.52±0.17b |
Akap12 | 3 | 1.03±0.25 | 0.19±0.13a | 0.76±0.25b |
DLL1 | 3 | 1.00±0.11 | 0.32±0.07a | 0.58±0.06b |
NR4A2 | 3 | 1.06±0.40 | 3.28±0.50a | 2.25±0.49b |
TFRC | 3 | 1.01±0.18 | 2.13±0.29a | 1.64±0.22b |
Table 1 qRT-PCR validation of miRNAs expression
Group | n | Normal | Model | XLHZ |
---|---|---|---|---|
miR-20a-3p | 3 | 1.08±0.49 | 6.65±1.38a | 3.31±0.76b |
miR-320-3p | 3 | 1.04±0.31 | 0.15±0.06a | 0.44±0.15b |
miR-34b-5p | 3 | 1.05±0.37 | 0.11±0.07a | 0.58±0.13c |
miR-483-3p | 3 | 1.01±0.15 | 0.29±0.16a | 0.37±0.12 |
miR-883-3p | 3 | 1.03±0.28 | 0.21±0.05a | 0.52±0.17b |
Akap12 | 3 | 1.03±0.25 | 0.19±0.13a | 0.76±0.25b |
DLL1 | 3 | 1.00±0.11 | 0.32±0.07a | 0.58±0.06b |
NR4A2 | 3 | 1.06±0.40 | 3.28±0.50a | 2.25±0.49b |
TFRC | 3 | 1.01±0.18 | 2.13±0.29a | 1.64±0.22b |
Figure 2 Effect of XLHZ on proliferation and migration of MNNG-induced GES-1 cells A: EdU assay was used to detect cell proliferation (magnification × 200). The proliferating cells were fluorescently stained with EdU (Green). The nuclei were stained with DAPI (Blue). the higher the cell proliferation rate, the brighter the green in the plot. A1: EdU of the control group; A2: DAPI of the control group; A3: merge of the control group; A4: EdU of the model group; A5: DAPI of the model group; A6: merge of the model group; A7: EdU of the XLHZ group; A8: DAPI of the XLHZ group; A9: merge of the XLHZ group. B: Quantitative analysis of proliferating cells. C: Wound healing assay was used to detect cell migration in the normal, model and XLHZ groups (magnification × 200). C1-C5: The migration of cells in the normal group was observed at 0, 12, 24, 48, 72 h. C6-C10: The migration of cells in the model group was observed at 0, 12, 24, 48, 72 h. C11-C15: The migration of cells in the XLHZ group was observed at 0, 12, 24, 48, 72 h. D: quantitative analysis of cells migration rate at 0, 12, 24, 48, 72 h. normal group: drink and eat freely + the same volume of normal saline was given by gavage; model group: MNNG, 180 μg/mL + 2% sodium salicylate, 1 mL/100 g per day + hunger and satiety disorders; XLHZ: model group + XLHZ, 36 g·kg-1·d-1. XLHZ: Xianglian Huazhuo formula; MNNG: N-Methyl-N'-Nitro-N-Nitrosoguanidine; GES-1: human gastric mucosal epithelial cells; EdU: 5-Ethynyl-2’- deoxyuridine; DAPI: 4’,6-Diamidino-2’-phenylindole. Statistical analyses were measured using one-way analysis of variance followed by post hoc Bonferroni correction for multiple comparisons. Data were presented as mean ± standard deviation (n = 3). Compared with the normal group, aP < 0.05, bP < 0.01; Compared with the model group, cP < 0.05; dP < 0.01.
Figure 3 Effect of XLHZ on apoptosis and cell cycle of MNNG-induced GES-1 cells A: flow cytometry assay was used to detect apoptosis in the normal, model and XLHZ groups. A1: normal group; A2: model group; A3: XLHZ group; A4: apoptosis rate of quantitative analysis. B: flow cytometry assay was used to detect cell cycle in the normal, model and XLHZ groups. B1: normal group; B2: model group; B3: XLHA group; B4: quantitative analysis of cell cycle distribution. normal group: drink and eat freely + the same volume of normal saline was given by gavage; model group: MNNG, 180 μg/mL + 2% sodium salicylate, 1 mL/100 g per day + hunger and satiety disorders; XLHZ: model group + XLHZ, 36 g·kg-1·d-1. XLHZ: Xianglian Huazhuo formula; MNNG: N-Methyl-N'-Nitro-N-Nitrosoguanidine; GES-1: human gastric mucosal epithelial cells. Statistical analyses were measured using one-way analysis of variance followed by post hoc Bonferroni correction for multiple comparisons. Data were presented as mean ± standard deviation (n = 3). Compared with the normal group, aP < 0.01; Compared with the model group, bP < 0.01.
Figure 4 Effects of XLHZ on EMT and proliferation of MNNG-induced GES-1 cells A: representative images of of E-cadherin, TGF-β1, vimentin and β-catenin protein expression that were exhibited from MNNG-induced GES-1 cells in the normal, model and XLHZ groups. B: quantitative analysis of E-cadherin protein; C: quantitative analysis of TGF-β1 protein; D: quantitative analysis of vimentin protein; E: quantitative analysis of β-catenin protein. normal group: drink and eat freely + the same volume of normal saline was given by gavage; model group: MNNG, 180 μg/mL + 2% sodium salicylate, 1 mL/100 g per day + hunger and satiety disorders; XLHZ: model group + XLHZ, 36 g·kg-1·d-1. XLHZ: Xianglian Huazhuo formula; EMT: epithelial mesenchymal transition; MNNG: N-Methyl-N'-Nitro-N-Nitrosoguanidine; GES-1: human gastric mucosal epithelial cells. Statistical analyses were measured using one-way analysis of variance followed by post hoc Bonferroni correction for multiple comparisons. Data were presented as mean ± standard deviation (n = 3). Compared with the normal group, aP < 0.01; Compared with the model group, bP < 0.01; cP < 0.05.
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