Journal of Traditional Chinese Medicine ›› 2025, Vol. 45 ›› Issue (6): 1263-1272.DOI: 10.19852/j.cnki.jtcm.2025.06.007
• Original Articles • Previous Articles Next Articles
Effect of Xuanfei Tongqiao acupuncture on nasal inflammation in rats with allergic rhinitis: modulation of long non-coding RNA growth arrest-specific transcript 5 methylation modification
LYU Hequn1,2, ZENG Chunli3,4, ZHANG Hanrui5, YANG Chen6, SHEN Yan6(
), PENG Yongjun7,8()
- 1 Department of Acupuncture and Rehabilitation, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, China
2 Department of Acupuncture, Yancheng TCM Hospital Affiliated to Nanjing University of Chinese Medicine, Yancheng 224000, China
3 Department of Lung Disease, Yancheng TCM Hospital Affiliated to Nanjing University of Chinese Medicine, Yancheng 224000, China
4 Department of Critical Care Medicine, Seventh People’s Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai 200137, China
5 Department of Acupuncture and Rehabilitation, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, China
6 National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300000, China
7 Department of Acupuncture and Rehabilitation, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, China
8 National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300000, China
-
Received:2024-06-12Accepted:2024-12-30Online:2025-12-15Published:2025-11-24 -
Contact:SHEN Yan, National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300000, China. swshtj@163.com, Telephone: + 86-13770671498; + 86-18920396086;
Dr. PENG Yongjun, Department of Acupuncture and Rehabilitation, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, China. pengyongjun2004@126.com -
About author:LYU Hequn and ZENG Chunli are co-first authors and contributed equally to this work -
Supported by:Multi-Center Clinical Study on Xuanfei Tongqiao Acupuncture combined with Sphenopalatine Ganglion Stimulation in the Treatment of Allergic Rhinitis(NCRCOP2023008);Clinical Observation on Preventive Acupuncture for Seasonal Allergic Rhinitis based on the Theory of "Strong Spleen Prevents Pathogenic Invasion"(2020043);Mechanism of Xuanfei Tongqiao Acupuncture in Treating Allergic Rhinitis via m6A Methylation Modification of lncRNA GAS5(YK2023017)
Cite this article
LYU Hequn, ZENG Chunli, ZHANG Hanrui, YANG Chen, SHEN Yan, PENG Yongjun. Effect of Xuanfei Tongqiao acupuncture on nasal inflammation in rats with allergic rhinitis: modulation of long non-coding RNA growth arrest-specific transcript 5 methylation modification[J]. Journal of Traditional Chinese Medicine, 2025, 45(6): 1263-1272.
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Figure 1 Comparison of behavioral scores among different groups of rats before and after intervention Control group: received intraperitoneal and intranasal saline administration; AR group: OVA-sensitized allergic rhinitis model group; AR + Acupuncture group: AR model plus acupuncture treatment; AR + sh-GAS5 group: AR model plus treatment with lentivirus expressing short hairpin RNA targeting lncRNA-GAS5. AR: allergic rhinitis; sh-GAS5: short hairpin RNA-GAS5. Statistical analyse were meansured using one-way analysis of variance followed by post hoc Bonfferroni correction for multiple comparisons. Data were presented as mean ± standard deviation (n = 6). Statistical analysis was performed using one-way analysis of variance followed by post hoc Bonferroni correction for multiple comparisons. Compared with the Control group, aP < 0.05; compared with the AR group, bP < 0.05; compared with the AR + Acupuncture group, cP < 0.05.
Figure 1 Comparison of behavioral scores among different groups of rats before and after intervention Control group: received intraperitoneal and intranasal saline administration; AR group: OVA-sensitized allergic rhinitis model group; AR + Acupuncture group: AR model plus acupuncture treatment; AR + sh-GAS5 group: AR model plus treatment with lentivirus expressing short hairpin RNA targeting lncRNA-GAS5. AR: allergic rhinitis; sh-GAS5: short hairpin RNA-GAS5. Statistical analyse were meansured using one-way analysis of variance followed by post hoc Bonfferroni correction for multiple comparisons. Data were presented as mean ± standard deviation (n = 6). Statistical analysis was performed using one-way analysis of variance followed by post hoc Bonferroni correction for multiple comparisons. Compared with the Control group, aP < 0.05; compared with the AR group, bP < 0.05; compared with the AR + Acupuncture group, cP < 0.05.
Figure 2 Histopathological evaluation of nasal mucosa in rat models of AR and treatment groups by HE staining A: control group exhibits normal nasal mucosal architecture composed of pseudostratified epithelial cells; A1: intact glandular structures; A2: no inflammatory cell infiltration; A3: no goblet cells observed; B: AR group demonstrates characteristic pathological alterations; B1: thickened mucosal layer; B2: increased goblet cell count; B3: inflammatory cell infiltration with eosinophils; C: AR + Acupuncture group shows attenuated inflammation and reduced epithelial hyperplasia; C1: thinner mucosal layer; C2: decreased goblet cell count; C3: reduced inflammatory cell infiltration and eosinophils; D: AR + sh-GAS5 group exhibits therapeutic effects comparable to the Acupuncture group; D1: improved mucosal thickness; D2: reduced goblet cell number; D3: decreased inflammatory cell infiltration. Control group: received intraperitoneal and intranasal saline administration; AR group: OVA-sensitized allergic rhinitis model group; AR + Acupuncture group: AR model plus acupuncture treatment; AR + sh-GAS5 group: AR model plus treatment with lentivirus expressing short hairpin RNA targeting lncRNA-GAS5. AR: allergic rhinitis; HE: hematoxylin/eosin; OVA: ovalbumin; GAS5: growth arrest-specific transcript 5; sh-GAS5: short hairpin RNA-GAS5. Scale bar = 20 μm (applies to all panels).
Figure 2 Histopathological evaluation of nasal mucosa in rat models of AR and treatment groups by HE staining A: control group exhibits normal nasal mucosal architecture composed of pseudostratified epithelial cells; A1: intact glandular structures; A2: no inflammatory cell infiltration; A3: no goblet cells observed; B: AR group demonstrates characteristic pathological alterations; B1: thickened mucosal layer; B2: increased goblet cell count; B3: inflammatory cell infiltration with eosinophils; C: AR + Acupuncture group shows attenuated inflammation and reduced epithelial hyperplasia; C1: thinner mucosal layer; C2: decreased goblet cell count; C3: reduced inflammatory cell infiltration and eosinophils; D: AR + sh-GAS5 group exhibits therapeutic effects comparable to the Acupuncture group; D1: improved mucosal thickness; D2: reduced goblet cell number; D3: decreased inflammatory cell infiltration. Control group: received intraperitoneal and intranasal saline administration; AR group: OVA-sensitized allergic rhinitis model group; AR + Acupuncture group: AR model plus acupuncture treatment; AR + sh-GAS5 group: AR model plus treatment with lentivirus expressing short hairpin RNA targeting lncRNA-GAS5. AR: allergic rhinitis; HE: hematoxylin/eosin; OVA: ovalbumin; GAS5: growth arrest-specific transcript 5; sh-GAS5: short hairpin RNA-GAS5. Scale bar = 20 μm (applies to all panels).
Figure 3 Flow cytometric characterization and quantification of ILC2 populations in AR models A: control group gating strategy; A1: gating of CD45+ leukocytes (PE-A) from live single cells; A2: identification of Lineage? (PB450-A) subsets in CD45+ population; A3: ILC2s defined as CD25+ (APC-A) in CD45+ lineage? cells; B: AR group gating strategy; B1: gating of CD45+ leukocytes (PE-A) from live single cells; B2: lineage? (PB450-A) subsets identified in CD45+ leukocytes; B3: significantly increased ILC2 frequency compared to the control group; C: AR + acupuncture group gating strategy; C1: gating of CD45+ leukocytes (PE-A) from live single cells; C2: lineage? (PB450-A) subsets in CD45+ population; C3: marked reduction in ILC2 frequency compared to the AR group; D: AR + sh-GAS5 group gating strategy; D1: gating of CD45+ leukocytes (PE-A) from live single cells; D2: lineage? (PB450-A) subsets resolved from CD45? leukocytes; D3: significant decrease in ILC2 abundance relative to the AR group; E: ILC2s frequency expressed as percentage of live cells. Control group: received intraperitoneal and intranasal saline administration; AR group: OVA-sensitized allergic rhinitis model group; AR + Acupuncture group: AR model plus acupuncture treatment; AR+sh-GAS5 group: AR model plus treatment with lentivirus expressing short hairpin RNA targeting lncRNA-GAS5. AR: allergic rhinitis; sh-GAS5: short hairpin RNA-GAS5; PE: phycoerythrin; GAS5: growth arrest-specific transcript 5; ILC2: type 2 innate lymphoid cells; OVA: ovalbumin. Data are presented as mean ± standard deviation (n = 3). Statistical analysis was performed using one-way analysis of variance followed by post hoc Bonferroni correction for multiple comparisons. Compared with the Control group, aP < 0.05; compared with the AR group, bP < 0.05; compared with the AR + Acupuncture group, cP < 0.05.
Figure 3 Flow cytometric characterization and quantification of ILC2 populations in AR models A: control group gating strategy; A1: gating of CD45+ leukocytes (PE-A) from live single cells; A2: identification of Lineage? (PB450-A) subsets in CD45+ population; A3: ILC2s defined as CD25+ (APC-A) in CD45+ lineage? cells; B: AR group gating strategy; B1: gating of CD45+ leukocytes (PE-A) from live single cells; B2: lineage? (PB450-A) subsets identified in CD45+ leukocytes; B3: significantly increased ILC2 frequency compared to the control group; C: AR + acupuncture group gating strategy; C1: gating of CD45+ leukocytes (PE-A) from live single cells; C2: lineage? (PB450-A) subsets in CD45+ population; C3: marked reduction in ILC2 frequency compared to the AR group; D: AR + sh-GAS5 group gating strategy; D1: gating of CD45+ leukocytes (PE-A) from live single cells; D2: lineage? (PB450-A) subsets resolved from CD45? leukocytes; D3: significant decrease in ILC2 abundance relative to the AR group; E: ILC2s frequency expressed as percentage of live cells. Control group: received intraperitoneal and intranasal saline administration; AR group: OVA-sensitized allergic rhinitis model group; AR + Acupuncture group: AR model plus acupuncture treatment; AR+sh-GAS5 group: AR model plus treatment with lentivirus expressing short hairpin RNA targeting lncRNA-GAS5. AR: allergic rhinitis; sh-GAS5: short hairpin RNA-GAS5; PE: phycoerythrin; GAS5: growth arrest-specific transcript 5; ILC2: type 2 innate lymphoid cells; OVA: ovalbumin. Data are presented as mean ± standard deviation (n = 3). Statistical analysis was performed using one-way analysis of variance followed by post hoc Bonferroni correction for multiple comparisons. Compared with the Control group, aP < 0.05; compared with the AR group, bP < 0.05; compared with the AR + Acupuncture group, cP < 0.05.
Figure 4 Comparative analysis of m6A-related and type 2 inflammation markers in rat nasal mucosa across experimental groups A: representative Western blot bands showing protein expression of METTL3, GATA3, IL-33, and GAPDH; B: quantitative analysis of METTL3 protein expression normalized to GAPDH; C: quantitative analysis of GATA3 protein expression normalized to GAPDH; D: quantitative analysis of IL-33 protein expression normalized to GAPDH; E: lncRNA-GAS5 expression levels across experimental groups; F: METTL3 mRNA expression levels across experimental groups. Control group: received intraperitoneal and intranasal saline administration; AR group: OVA-sensitized allergic rhinitis model group; AR + Acupuncture group: AR model plus acupuncture treatment; AR + sh-GAS5 group: AR model plus treatment with lentivirus expressing short hairpin RNA targeting lncRNA-GAS5. ILC2s: type 2 innate lymphoid cells; GATA3: GATA binding protein 3; METTL3: methyltransferase-like 3; IL-33: interleukin-33; GAPDH: glyceraldehyde-3-phosphate dehydrogenase. AR: allergic rhinitis; sh-GAS5: short hairpin RNA-GAS5; METTL3: methyltransferase-like 3; GAS5: growth arrest-specific transcript 5; OVA: ovalbumin. Data are presented as mean ± standard deviation (n = 3). Statistical analysis was performed using one-way analysis of variance followed by post hoc Bonferroni correction for multiple comparisons. Compared with the Control group, aP < 0.05; compared with the AR group, bP < 0.05.
Figure 4 Comparative analysis of m6A-related and type 2 inflammation markers in rat nasal mucosa across experimental groups A: representative Western blot bands showing protein expression of METTL3, GATA3, IL-33, and GAPDH; B: quantitative analysis of METTL3 protein expression normalized to GAPDH; C: quantitative analysis of GATA3 protein expression normalized to GAPDH; D: quantitative analysis of IL-33 protein expression normalized to GAPDH; E: lncRNA-GAS5 expression levels across experimental groups; F: METTL3 mRNA expression levels across experimental groups. Control group: received intraperitoneal and intranasal saline administration; AR group: OVA-sensitized allergic rhinitis model group; AR + Acupuncture group: AR model plus acupuncture treatment; AR + sh-GAS5 group: AR model plus treatment with lentivirus expressing short hairpin RNA targeting lncRNA-GAS5. ILC2s: type 2 innate lymphoid cells; GATA3: GATA binding protein 3; METTL3: methyltransferase-like 3; IL-33: interleukin-33; GAPDH: glyceraldehyde-3-phosphate dehydrogenase. AR: allergic rhinitis; sh-GAS5: short hairpin RNA-GAS5; METTL3: methyltransferase-like 3; GAS5: growth arrest-specific transcript 5; OVA: ovalbumin. Data are presented as mean ± standard deviation (n = 3). Statistical analysis was performed using one-way analysis of variance followed by post hoc Bonferroni correction for multiple comparisons. Compared with the Control group, aP < 0.05; compared with the AR group, bP < 0.05.
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