Journal of Traditional Chinese Medicine ›› 2025, Vol. 45 ›› Issue (1): 1-12.DOI: 10.19852/j.cnki.jtcm.2025.01.001
Hewei Jiangni granule (和胃降逆颗粒) alleviates visceral hypersensitivity of non-erosive reflux disease via stromal interaction molecule 1/transient receptor potential vanilloid subfamily member 1 pathway
CHENG Yuan1, ZHANG Xiaosi2, LI Junxiang2, ZHANG Liming2, DAI Yi2,3, XIE Chune2, SHI Lei2, LI Xiaohong2(
), KOU Fushun4()
- 1 School of Traditional Chinese Medicine and School of Integrated Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, China; Gastroenterology Department, Dongfang Hospital, Beijing University of Chinese Medicine, Beijing 100078, China
2 Gastroenterology Department, Dongfang Hospital, Beijing University of Chinese Medicine, Beijing 100078, China
3 Department of Pharmacotherapy and Oriental Medicine, School of Pharmacy, Hyogo University of Health Sciences, Hyogo 650-8530, Japan
4 Center for IBD Research, Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai 200072, China
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Received:2023-06-11Accepted:2023-10-22Online:2025-02-15Published:2025-01-10 -
Contact:Prof. LI Xiaohong, Gastroenterology Department, Dongfang Hospital, Beijing University of Chinese Medicine, Beijing 100078, China,lxhktxz@163.com ; Dr KOU Fushun, Center for IBD Research, Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai 200072, China,koufushun@126.com , Telephone: +86-10-67689655; +86-25-85311836 -
Supported by:National Natural Science Foundation of China: Study on the Molecular Mechanism of the Regulation of Crypt Goblet Cell Pyroptosis and Exocytosis to Repair Ulcerative Colitis Mucus Barrier by the Method of Clearing and Opening the Xuanfu from the Perspective of "Xuanfu-Crypt"(82305143);National Natural Science Foundation of China: Exploring the Molecular Mechanism of "Hewei Jiangni Fang" Intervention in Non-erosive Reflux Disease Esophageal Hypersensitivity from the Perspective of Mas-related Gene X2/Stromal Interaction Molecule 1/Cell Adhesion Molecule 1 Pathway Regulation of Mast Cell/Dorsal Root Ganglion Communication based on the "Xinkai-Kujiang" Method(82374401)
Cite this article
CHENG Yuan, ZHANG Xiaosi, LI Junxiang, ZHANG Liming, DAI Yi, XIE Chune, SHI Lei, LI Xiaohong, KOU Fushun. Hewei Jiangni granule (和胃降逆颗粒) alleviates visceral hypersensitivity of non-erosive reflux disease via stromal interaction molecule 1/transient receptor potential vanilloid subfamily member 1 pathway[J]. Journal of Traditional Chinese Medicine, 2025, 45(1): 1-12.
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Figure 1 HWJNG ameliorated pathological damage and regulated related indicators in NERD mice A: HE staining of esophageal mucosa in lower third of esophagus of mice (× 200), respectively; B: toluidine blue staining of esophageal mucosa in lower third of esophagus of mice (× 200), respectively; C: total counts of MCs; D: blood glucose; E: weights; F: PH values; G: mechanical paw withdrawal threshold; H: thermal withdrawal latency. A1, B1: Control group; A2, B2: Model group; A3, B3: Omeprazole group; A4, B4: HWJNG-L group; A5, B5: HWJNG-H group. Control group: deionized water; Model group: fructose water and restraint stress; Omeprazole group: model group + Omeprazole at 0.06 mg·kg-1·d-1; HWJNG-L: low dose of HWJNG at 0.24 g/kg; HWJNG-H: high dose of HWJNG at 0.48 g/kg. HWJNG: Hewei Jiangni granule. Statistical analyses were measured using one-way analysis of variance for multimal comparisons. Data were presented as mean ± standard deviation (n = 5). Compared with the control group, aP < 0.001, dP < 0.01; compared with the model group, bP < 0.001, eP < 0.01; compared with the HWJNG-H group, cP = 0.08, fP < 0.05.
Figure 1 HWJNG ameliorated pathological damage and regulated related indicators in NERD mice A: HE staining of esophageal mucosa in lower third of esophagus of mice (× 200), respectively; B: toluidine blue staining of esophageal mucosa in lower third of esophagus of mice (× 200), respectively; C: total counts of MCs; D: blood glucose; E: weights; F: PH values; G: mechanical paw withdrawal threshold; H: thermal withdrawal latency. A1, B1: Control group; A2, B2: Model group; A3, B3: Omeprazole group; A4, B4: HWJNG-L group; A5, B5: HWJNG-H group. Control group: deionized water; Model group: fructose water and restraint stress; Omeprazole group: model group + Omeprazole at 0.06 mg·kg-1·d-1; HWJNG-L: low dose of HWJNG at 0.24 g/kg; HWJNG-H: high dose of HWJNG at 0.48 g/kg. HWJNG: Hewei Jiangni granule. Statistical analyses were measured using one-way analysis of variance for multimal comparisons. Data were presented as mean ± standard deviation (n = 5). Compared with the control group, aP < 0.001, dP < 0.01; compared with the model group, bP < 0.001, eP < 0.01; compared with the HWJNG-H group, cP = 0.08, fP < 0.05.
Figure 2 Observation of pathological damage in non-erosive reflux disease repaired by HWJNG under electron microscopy A: dilated epithelial cell space (× 1000); B: dilated intercellular space (× 2500); C: mitochondria morphological changes (× 8000); D: desmosome morphological changes (× 15 000); E: width of mucosal intercellular space (× 2500); F: quantity of desmosomes (× 15000). A1, B1, C1, D1: Control group; A2, B2, C2, D2: model group; A3, B3, C3, D3: omeprazole group; A4, B4, C4, D4: HWJNG-L group; A5, B5, C5, D5: HWJNG-H group. Control group: deionized water; Model group: fructose water and restraint stress; Omeprazole group: model group + Omeprazole at 0.06 mg·kg-1·d-1; HWJNG-L: low dose of HWJNG at 0.24 g/kg; HWJNG-H: high dose of HWJNG at 0.48 g/kg. HWJNG: Hewei Jiangni granule. Statistical analyses were measured using one-way analysis of variance for multimal comparisons. Data were presented as mean ± standard deviation (n = 5). Compared with the control group, aP < 0.001; compared with the model group, bP < 0.001; compared with the HWJNG-H group, cP < 0.05.
Figure 2 Observation of pathological damage in non-erosive reflux disease repaired by HWJNG under electron microscopy A: dilated epithelial cell space (× 1000); B: dilated intercellular space (× 2500); C: mitochondria morphological changes (× 8000); D: desmosome morphological changes (× 15 000); E: width of mucosal intercellular space (× 2500); F: quantity of desmosomes (× 15000). A1, B1, C1, D1: Control group; A2, B2, C2, D2: model group; A3, B3, C3, D3: omeprazole group; A4, B4, C4, D4: HWJNG-L group; A5, B5, C5, D5: HWJNG-H group. Control group: deionized water; Model group: fructose water and restraint stress; Omeprazole group: model group + Omeprazole at 0.06 mg·kg-1·d-1; HWJNG-L: low dose of HWJNG at 0.24 g/kg; HWJNG-H: high dose of HWJNG at 0.48 g/kg. HWJNG: Hewei Jiangni granule. Statistical analyses were measured using one-way analysis of variance for multimal comparisons. Data were presented as mean ± standard deviation (n = 5). Compared with the control group, aP < 0.001; compared with the model group, bP < 0.001; compared with the HWJNG-H group, cP < 0.05.
Figure 3 HWJNG treatment regulates protein and mRNA expressions of STIM1, TRPV1 and related neurotransmitters in esophageal mucosa and DRG of mice A: relative protein levels of SP, PAR2, and CGRP in esophageal mucosa (n = 5). A1: SP; A2: PAR2; A3: CGRP. B: relative m RNA levels of SP, PAR2 and CGRP assessed in esophageal mucosa (n = 3). B1: SP; B2: PAR2; B3: CGRP. C: relativemRNA levels of SP, CGRP, STIM1 and TRPV1 assessed in DRG (n = 3). C1: SP; C2: CGRP; C3: STIM1; C4: TRPV1. D: representative band of STIM1 and TRPV1 protein detected by WB in DRG (n = 3). E: relative protein levels of STIM1 results in DRG (n = 3). F: relative protein levels of TRPV1 results in DRG (n = 3). Control group: deionized water; Model group: fructose water and restraint stress; HWJNG: Low dose of HWJNG at 0.24 g/kg + high dose of HWJNG at 0.48 g/kg. STIM1: stromal interaction molecule 1; TRPV1: transient receptor potential vanilloid subfamily member 1; DRG: dorsal root ganglion cells; SP: substance P; PAR2: protease activated receptors-2; CGRP: calcitonin gene-related peptide; HWJNG: Hewei Jiangni granule. Statistical analyses were measured using one-way analysis of variance for multimal comparisons. Data were presented as mean ± standard deviation (n = 5). Compared with the control group, aP < 0.001, cP < 0.01; compared with the model group, bP < 0.001, dP < 0.01. eP < 0.05.
Figure 3 HWJNG treatment regulates protein and mRNA expressions of STIM1, TRPV1 and related neurotransmitters in esophageal mucosa and DRG of mice A: relative protein levels of SP, PAR2, and CGRP in esophageal mucosa (n = 5). A1: SP; A2: PAR2; A3: CGRP. B: relative m RNA levels of SP, PAR2 and CGRP assessed in esophageal mucosa (n = 3). B1: SP; B2: PAR2; B3: CGRP. C: relativemRNA levels of SP, CGRP, STIM1 and TRPV1 assessed in DRG (n = 3). C1: SP; C2: CGRP; C3: STIM1; C4: TRPV1. D: representative band of STIM1 and TRPV1 protein detected by WB in DRG (n = 3). E: relative protein levels of STIM1 results in DRG (n = 3). F: relative protein levels of TRPV1 results in DRG (n = 3). Control group: deionized water; Model group: fructose water and restraint stress; HWJNG: Low dose of HWJNG at 0.24 g/kg + high dose of HWJNG at 0.48 g/kg. STIM1: stromal interaction molecule 1; TRPV1: transient receptor potential vanilloid subfamily member 1; DRG: dorsal root ganglion cells; SP: substance P; PAR2: protease activated receptors-2; CGRP: calcitonin gene-related peptide; HWJNG: Hewei Jiangni granule. Statistical analyses were measured using one-way analysis of variance for multimal comparisons. Data were presented as mean ± standard deviation (n = 5). Compared with the control group, aP < 0.001, cP < 0.01; compared with the model group, bP < 0.001, dP < 0.01. eP < 0.05.
Figure 4 STIM1 deficiency and overexpression affect the therapeutic of HWJNG on DRG cells with P815 cells co-culture A: P815 cells adhesion to DRGs cells via a fluorescence adhesion assay in different groups. The protein levels of PAR2. A1: Blank; A2: Model; A3: model + HWJNG; A4: model + oeCtrl; A5: model + oeSTIM1 + HWJNG; A6: model + siCtrl; A7: model + siSTIM1. B: relative protein levels of SP assessed by ELISA in DRG cells. C: relative protein levels of PAR2 assessed by ELISA in DRG cells. D: relative protein levels of CGRP assessed by ELISA in DRG cells. E: levels of Orai1 mRNA assessed by RT- qPCR in DRG cells. F: levels of TRPV1 mRNA assessed by RT- qPCR in DRG cells. G: levels of STIM1 mRNA assessed by RT- qPCR in DRG cells. Blank group: DRG cells with P815 Cells; Model group: blank group + SP; Model + HWJNG: Model group + HWJNG; Model + oeCtrl: Model group + oeCtrl; Model + oeSTIM1 + HWJNG; Model group + oeSTIM1 + HWJNG; Model + siCtrl: Model group + siCtrl; Model + siSTIM1: Model group + siSTIM1; oeCtrl; STIM1 overexpression control; oeSTIMI; overexpression STIM1; siCtrl; STIM1 siRNA control; siSTIM1; siRNA STIM1. SP: substance P; STIM1: stromal interaction molecule 1; TRPV1: potential vanilloid subfamily member 1; PAR-2: protease activated receptor-2; CGRP: alcitonin gene-related peptide; HWJNG: Hewei Jiangni granule. Statistical analyses were measured using one-way analysis of variance for multimal comparisons. Data were presented as mean ± standard deviation (n = 5). Compared with the control group, aP < 0.001; compared with the model group, bP < 0.001; compared with the model + HWJNG group, cP < 0.01, dP < 0.05.
Figure 4 STIM1 deficiency and overexpression affect the therapeutic of HWJNG on DRG cells with P815 cells co-culture A: P815 cells adhesion to DRGs cells via a fluorescence adhesion assay in different groups. The protein levels of PAR2. A1: Blank; A2: Model; A3: model + HWJNG; A4: model + oeCtrl; A5: model + oeSTIM1 + HWJNG; A6: model + siCtrl; A7: model + siSTIM1. B: relative protein levels of SP assessed by ELISA in DRG cells. C: relative protein levels of PAR2 assessed by ELISA in DRG cells. D: relative protein levels of CGRP assessed by ELISA in DRG cells. E: levels of Orai1 mRNA assessed by RT- qPCR in DRG cells. F: levels of TRPV1 mRNA assessed by RT- qPCR in DRG cells. G: levels of STIM1 mRNA assessed by RT- qPCR in DRG cells. Blank group: DRG cells with P815 Cells; Model group: blank group + SP; Model + HWJNG: Model group + HWJNG; Model + oeCtrl: Model group + oeCtrl; Model + oeSTIM1 + HWJNG; Model group + oeSTIM1 + HWJNG; Model + siCtrl: Model group + siCtrl; Model + siSTIM1: Model group + siSTIM1; oeCtrl; STIM1 overexpression control; oeSTIMI; overexpression STIM1; siCtrl; STIM1 siRNA control; siSTIM1; siRNA STIM1. SP: substance P; STIM1: stromal interaction molecule 1; TRPV1: potential vanilloid subfamily member 1; PAR-2: protease activated receptor-2; CGRP: alcitonin gene-related peptide; HWJNG: Hewei Jiangni granule. Statistical analyses were measured using one-way analysis of variance for multimal comparisons. Data were presented as mean ± standard deviation (n = 5). Compared with the control group, aP < 0.001; compared with the model group, bP < 0.001; compared with the model + HWJNG group, cP < 0.01, dP < 0.05.
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