Regulating the Balance of Th17/Treg via Electroacupuncture and Moxibustion: An Ulcerative Colitis Mice Model Based Study - PubMed (original) (raw)

doi: 10.1155/2017/7296353. Epub 2017 Dec 17.

Hejiaozi Zhang 1 3, Chengyulin Wang 1, Mingxiao Yang 1, Shyang Chang 4, Yu Geng 1, Hui Yang 1, Zhiqi Zhuang 1, Xiang Wang 1, Lushuang Xie 1, Biao Huang 1, Na Zhao 1, Wei Zhou 1, Xinhui Cheng 5, Bei Cai 6, Qiaofeng Wu 1, Shu Guang Yu 1

Affiliations

Regulating the Balance of Th17/Treg via Electroacupuncture and Moxibustion: An Ulcerative Colitis Mice Model Based Study

Jungang Sun et al. Evid Based Complement Alternat Med. 2017.

Abstract

Aim: To investigate the relationship between the effects of electroacupuncture/moxibustion and the balance of Th17/Treg in treating ulcerative colitis (UC) and to preliminary compare the effects of the above two methods.

Methods: DSS-induced UC mice were treated by electroacupuncture and moxibustion. Disease activity index (DAI) was scored; intestinal pathological structure and ultrastructure were observed. The levels of IL-2, IL-6, IL-10, IL-17A, IL-17F, and TGF-β in plasma were measured by ELISA. The percentages of Treg and Th17 in spleen lymphocytes were analyzed by flow cytometry. Also, the expressions of TLR2, TLR4, ROR_γ_t, and FOXP3 in the distal colon were detected by immunohistochemistry or western blot.

Results: Both electroacupuncture and moxibustion can relieve UC. These effects are further supported by ELISA results. In addition, the ratio of Treg and Th17 in spleen lymphocytes and the expression of TLR2 and TLR4 are significantly improved. Also, the expression of ROR_γ_t and FOXP3 in distal colon were improved. Besides, the effect of moxibustion is better than that of electroacupuncture on TLR2, TLR4, and FOXP3 expression (P < 0.05).

Conclusion: Both electroacupuncture and moxibustion may ameliorate UC by regulating the balance of Th17/Treg. Whether moxibustion has better efficacy than electroacupuncture needs further study.

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Figures

Figure 1

Figure 1

HE staining results of distal colonic tissues in different group. Normal mucosal epithelia, intestinal glands, and structure were seen in distal colonic tissues in the control group (a). Damaged mucosa, disordered glandular structure, and apparent edema in and underneath the mucosa were found in the DSS-induced model group mice (b). And the histopathological conditions have been greatly improved in acupuncture (c) and moxibustion group (d) (∗_normal architecture and lack of cellular infiltrate; #histological damage; ∇Globlet cell depletion; magnification, 200x, and scale bar = 100 μm_).

Figure 2

Figure 2

The ultrastructure inspection of the distal colon tissues by electron microscopy. Compared with the control group (a), the model group (b) has shown shortened loose microvilli with uneven length (M), large intraepithelial vacuoles (V), and unclear dissolved mucosal granules in the cytoplasm. After acupuncture (c) or moxibustion (d) treatment, the microvilli are in good order (magnification, 10000x, and scale bar = 2 μm).

Figure 3

Figure 3

Toll-like receptor (TLR) 2 and TLR4 were detected in the colon by the immunohistochemical method. Compared with the control group (a, e), the intestinal TLR2 and TLR4 expression in normal in model group have increased (b, f). After acupuncture (c, g) or moxibustion (d, h) treatment, the expressions of TLR2 and TLR4 have been reduced. The average IOD was obtained by analyzing TLR2 (i) and TLR4 (j) immunohistochemistry staining in five random fields of each slide. Compared with electroacupuncture group, the OD level of TLR2 and TLR4 in moxibustion group is lower. ∗ P < 0.05; ∗∗ P < 0.01 (magnification, 200x, and scale bar = 100 μm).

Figure 4

Figure 4

The protein expressions of TLR2 and TLR4 in the distal colon tissues. The results showed that the TLR2 and TLR4 protein expressions of the distal colon in model group were significantly enhanced, while, after acupuncture and moxibustion treatments, these two proteins were significantly reduced (P < 0.01). In addition, the level of TLR2 in moxibustion group is lower than that in the electroacupuncture group. ∗ P < 0.05; ∗∗ P < 0.01.

Figure 5

Figure 5

Flow cytometric pictures and the percentage of CD4+CD25+Foxp3+Treg cells and CD3+CD8+IL-17+Th17 cells among CD4+T cells in each group. Lymphocytes from spleen of each group were stained with labeled as anti-mice antibodies as in the described methods. Treg cells and Th17 were gated and shown in the right-upper quadrant. Compared with control group, the percentage of Treg (CD4+CD25+Foxp3+Treg/CD4+ T cells) in model group decreased (P < 0.05). Treg cells content increased after acupuncture or moxibustion treatment (P < 0.05). On the contrary, the percentage of Th17 cells significantly increased (P < 0.05) in UC model mice while decreasing by both acupuncture and moxibustion. ∗ P < 0.05.

Figure 6

Figure 6

Quantification of the key immune factors by ELISA test: TNF-β, IL-10, and IL-2; IL-6, IL-17A, and IL-17F. The results showed that the levels of anti-inflammatory factors including TGF-β, IL-10, and IL-2 in the UC model group significantly decreased (Figure 6, upper panel; as compared with control group P < 0.01). On the contrary, the levels of proinflammatory factors including IL-6, IL-17A, and IL-17F significantly increased after modeling (Figure 6, lower panel; as compared with control group P < 0.01). After acupuncture and moxibustion treatments, the levels of TGF-β, IL-10, and IL-2 were significantly increased (Figure 6, upper panel) and the levels of IL-6, IL-17A, and IL-17F were significantly reduced (lower panel). In addition, there is no obvious difference between them in all these inflammation factors. ∗ P < 0.05; ∗∗ P < 0.01.

Figure 7

Figure 7

The protein expressions of ROR_γ_t and FOXP3 in the distal colon tissues. Compared with controls, the ROR_γ_t expression was significantly increased and FOXp3+ expression was significantly decreased in the UC model group (P < 0.01). However, after acupuncture or moxibustion treatment, ROR_γ_t expression was obviously decreased (P < 0.01) and FOXP3 expression was increased (P < 0.01). In addition, moxibustion increased higher FOXP3 than electroacupuncture. ∗ P < 0.05; ∗∗ P < 0.01.

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