Journal of Traditional Chinese Medicine ›› 2022, Vol. 42 ›› Issue (5): 732-740.DOI: 10.19852/j.cnki.jtcm.20220719.001
• Research Articles • Previous Articles Next Articles
Electroacupuncture at Tianshu (ST25) and Zusanli (ST36) alleviates stress-induced irritable bowel syndrome in mice by modulating gut microbiota and corticotropin-releasing factor
SUN Mengzhu1, ZHANG Yujie1, SONG Yafang1, GUO Jing1, ZHAO Tingting1, WANG Yuhang1, PEI Lixia2(
), SUN Jianhua2()
- 1 Department of Acupuncture and Rehabilitation, the Affiliated Hospital of Nanjing University of Chinese Meidicne, Nanjing 210023, China
2 Department of Acupuncture and Rehabilitation, Jiangsu Province Hospital of Chinese Medicine, Nanjing 210029, China
-
Received:2021-07-06Accepted:2021-10-14Online:2022-10-15Published:2022-07-19 -
Contact:PEI Lixia,SUN Jianhua -
About author:Prof. PEI Lixia, Department of Acupuncture and Rehabilitation, Jiangsu Province Hospital of Chinese Medicine, Nanjing 210029, China. plx11801758@163.com Telephone: +86-13851402838; +86-13914722816; +86-18851182170
Prof. SUN Jianhua, Department of Acupuncture and Rehabilitation, Jiangsu Province Hospital of Chinese Medicine, Nanjing 210029, China. drjhsun@sina.com;
-
Supported by:Natural Science Foundation-funded Project: Mechanism of Electroacupuncture Regulating CRF-NLRP6 Inflammasome-Related Pathways in Intestinal Flora Immune Dialogue(81804193);Leading Talents of Jiangsu Provincial Administration of TCM-funded Project: Clinical Effect and Mechanism of Acupuncture and Moxibustion on Irritable Bowel Syndrome(SLJ0206);eak Talent of Jiangsu Province Hospital of Chinese Medicine-funded Project: Clinical Effect and Mechanism of Acupuncture and Moxibustion in the Treatment of Functional Gastrointestinal Diseases(y2018rc05);Research and Innovation Project for Postgraduates of Jiangsu Province-funded Project: Mechanism of Electroacupuncture Regulating Bile Acid Receptor to Alleviate Visceral Hypersensitivity in Irritable Bowel Syndrome(YCX20_1469)
Cite this article
SUN Mengzhu, ZHANG Yujie, SONG Yafang, GUO Jing, ZHAO Tingting, WANG Yuhang, PEI Lixia, SUN Jianhua. Electroacupuncture at Tianshu (ST25) and Zusanli (ST36) alleviates stress-induced irritable bowel syndrome in mice by modulating gut microbiota and corticotropin-releasing factor[J]. Journal of Traditional Chinese Medicine, 2022, 42(5): 732-740.
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Table 1 AWR scores at different stimulation pressures in normal, WAS and WAS+EA groups ($\bar{x}$ ± s)
| Groups | n | AWR scores | |||
|---|---|---|---|---|---|
| 15 mm Hg | 30 mm Hg | 45 mm Hg | 60 mm Hg | ||
| Normal | 10 | 0.57±0.39 | 1.40±0.44 | 2.03±0.46 | 3.00±0.57 |
| WAS | 10 | 1.30±0.33a | 2.27±0.49a | 2.80±0.59c | 3.70±0.43d |
| WAS+EA | 10 | 0.80±0.48b | 1.77±0.52b | 2.27±0.49b | 3.30±0.46 |
Table 1 AWR scores at different stimulation pressures in normal, WAS and WAS+EA groups ($\bar{x}$ ± s)
| Groups | n | AWR scores | |||
|---|---|---|---|---|---|
| 15 mm Hg | 30 mm Hg | 45 mm Hg | 60 mm Hg | ||
| Normal | 10 | 0.57±0.39 | 1.40±0.44 | 2.03±0.46 | 3.00±0.57 |
| WAS | 10 | 1.30±0.33a | 2.27±0.49a | 2.80±0.59c | 3.70±0.43d |
| WAS+EA | 10 | 0.80±0.48b | 1.77±0.52b | 2.27±0.49b | 3.30±0.46 |
Figure 1 α-diversity of the gut microbiota A: Chao1 index in normal, WAS and WAS + EA groups; B: Ace index in normal, WAS and WAS + EA groups; C: Simpson index in normal, WAS and WAS + EA groups; D: Shannon index in normal, WAS and WAS + EA groups. Normal group: without treatment; WAS group: subjected to WAS for 1 h daily on 10 consecutive days; WAS + EA group: subjected to WAS for 1 h daily on 10 consecutive days and EA at bilateral Tianshu (ST25) and Zusanli (ST36) for 7 consecutive days. WAS: water avoidance stress; EA: electroacupuncture. aP < 0.01 versus normal group; bP < 0.01 versus WAS group.
Figure 1 α-diversity of the gut microbiota A: Chao1 index in normal, WAS and WAS + EA groups; B: Ace index in normal, WAS and WAS + EA groups; C: Simpson index in normal, WAS and WAS + EA groups; D: Shannon index in normal, WAS and WAS + EA groups. Normal group: without treatment; WAS group: subjected to WAS for 1 h daily on 10 consecutive days; WAS + EA group: subjected to WAS for 1 h daily on 10 consecutive days and EA at bilateral Tianshu (ST25) and Zusanli (ST36) for 7 consecutive days. WAS: water avoidance stress; EA: electroacupuncture. aP < 0.01 versus normal group; bP < 0.01 versus WAS group.
Figure 2 Structure of the gut microbiota A, B: PCoA plots of the unweighted and weighted UniFrac distances in normal, WAS and WAS + EA groups; C: NMDS ordination in normal, WAS and WAS + EA groups. Stress < 0.2 shows that NMDS can accurately reflect the difference between samples; D-E: the relative abundances of the microbial communities for each group at the phylum and genus levels. Normal group: without treatment; WAS group: subjected to WAS for 1 h daily on 10 consecutive days; WAS+EA group: subjected to WAS for 1 h daily on 10 consecutive days and EA at bilateral Tianshu (ST25) and Zusanli (ST36) for 7 consecutive days. WAS: water avoidance stress; EA: electroacupuncture; PCoA: principal co-ordinate analysis; NMDS: non-metric multidimensional scaling.
Figure 2 Structure of the gut microbiota A, B: PCoA plots of the unweighted and weighted UniFrac distances in normal, WAS and WAS + EA groups; C: NMDS ordination in normal, WAS and WAS + EA groups. Stress < 0.2 shows that NMDS can accurately reflect the difference between samples; D-E: the relative abundances of the microbial communities for each group at the phylum and genus levels. Normal group: without treatment; WAS group: subjected to WAS for 1 h daily on 10 consecutive days; WAS+EA group: subjected to WAS for 1 h daily on 10 consecutive days and EA at bilateral Tianshu (ST25) and Zusanli (ST36) for 7 consecutive days. WAS: water avoidance stress; EA: electroacupuncture; PCoA: principal co-ordinate analysis; NMDS: non-metric multidimensional scaling.
Figure 3 Composition of the gut microbiota A-B: MetaStat analysis at the genus and order level in normal, WAS and WAS+EA groups; C: histogram of LEfSe in normal, WAS and WAS+EA groups; D: cladogram of LEfSe in normal, WAS and WAS+EA groups. LDA score > 3.0. The red, green, or blue shading in the cladogram depicts bacterial taxa that were significantly higher in either the normal, WAS or WAS+EA groups, as indicated. p: phylum; c: class; o: order; f: family; g: genus; s: species. Normal group: without treatment; WAS group: subjected to WAS for 1 h daily on 10 consecutive days; WAS+EA group: subjected to WAS for 1 h daily on 10 consecutive days and EA at bilateral Tianshu (ST25) and Zusanli (ST36) for 7 consecutive days. WAS: water avoidance stress; EA: electroacupuncture; LDA: linear discriminant analysis.
Figure 3 Composition of the gut microbiota A-B: MetaStat analysis at the genus and order level in normal, WAS and WAS+EA groups; C: histogram of LEfSe in normal, WAS and WAS+EA groups; D: cladogram of LEfSe in normal, WAS and WAS+EA groups. LDA score > 3.0. The red, green, or blue shading in the cladogram depicts bacterial taxa that were significantly higher in either the normal, WAS or WAS+EA groups, as indicated. p: phylum; c: class; o: order; f: family; g: genus; s: species. Normal group: without treatment; WAS group: subjected to WAS for 1 h daily on 10 consecutive days; WAS+EA group: subjected to WAS for 1 h daily on 10 consecutive days and EA at bilateral Tianshu (ST25) and Zusanli (ST36) for 7 consecutive days. WAS: water avoidance stress; EA: electroacupuncture; LDA: linear discriminant analysis.
Figure 4 Effect of EA on the expression of CRF in colon tissues (magnification, × 200) A: relative mRNA expression of CRF in normal, WAS and WAS + EA groups; B: relative fluorescence intensity of CRF in normal, WAS and WAS + EA groups; C1-C3: confocal images of CRF, DAPI and merge (CRF + DAPI) in normal group; C4-C6: confocal images of CRF, DAPI and merge (CRF + DAPI) in WAS group; C7-C9: confocal images of CRF, DAPI and merge (CRF + DAPI) in WAS + EA group. Micrographs represent labeling of CRF (green) in mice colon tissues. Nuclear staining was conducted with DAPI (blue). Data were mean ± standard deviation (n = 3/ group). Normal group: without treatment; WAS group: subjected to WAS for 1 h daily on 10 consecutive days; WAS + EA group: subjected to WAS for 1 h daily on 10 consecutive days and EA at bilateral Tianshu (ST25) and Zusanli (ST36) for 7 consecutive days. EA: electroacupuncture; WAS: water avoidance stress; CRF: corticotropin-releasing factor; DAPI: 4',6-diamidino-2-phenylindole. aP < 0.001, cP < 0.01 versus normal group; bP < 0.001, dP < 0.05 versus WAS group.
Figure 4 Effect of EA on the expression of CRF in colon tissues (magnification, × 200) A: relative mRNA expression of CRF in normal, WAS and WAS + EA groups; B: relative fluorescence intensity of CRF in normal, WAS and WAS + EA groups; C1-C3: confocal images of CRF, DAPI and merge (CRF + DAPI) in normal group; C4-C6: confocal images of CRF, DAPI and merge (CRF + DAPI) in WAS group; C7-C9: confocal images of CRF, DAPI and merge (CRF + DAPI) in WAS + EA group. Micrographs represent labeling of CRF (green) in mice colon tissues. Nuclear staining was conducted with DAPI (blue). Data were mean ± standard deviation (n = 3/ group). Normal group: without treatment; WAS group: subjected to WAS for 1 h daily on 10 consecutive days; WAS + EA group: subjected to WAS for 1 h daily on 10 consecutive days and EA at bilateral Tianshu (ST25) and Zusanli (ST36) for 7 consecutive days. EA: electroacupuncture; WAS: water avoidance stress; CRF: corticotropin-releasing factor; DAPI: 4',6-diamidino-2-phenylindole. aP < 0.001, cP < 0.01 versus normal group; bP < 0.001, dP < 0.05 versus WAS group.
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