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Regulatory effect of moxibustion for rats with ulcerative colitis on the macrophage functional phenotype protein of lung tissue

艾灸对溃疡性结肠炎大鼠肺组织巨噬细胞功能表型蛋白调节作用的研究

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Abstract

Objective

To observe the regulatory effect of moxibustion on the expression of CD86 and CD163, which are the important functional phenotypes of macrophage differentiation in lung tissue of ulcerative colitis (UC) rats. The mechanism of the regulatory effect of moxibustion for macrophage differentiation based on the key cytokine interferon-gamma (IFN-γ), tumor necrosis death factor-alpha (TNF-α), interleukin- 4 (IL-4) and interleukin-13 (IL-13) was also explored.

Methods

Forty SD rats were randomly divided into a normal group (NG), a model group (MG), a normal moxibustion group (NMG) and a smokeless moxibustion group (SMG). The model of UC was made by antigen immunization combined with enema with topical formalin. The rats in the normal moxibustion group accepted moxibustion at bilateral Tianshu (ST 25), while the rats in the smokeless moxibustion group with smokeless moxibustion at bilateral Tianshu (ST 25), each 10 min, once a day for 8 d. After treatments, the hematoxylin-eosin (HE) staining was used to observe the pathological changes of colonic tissue, the Western blotting (WB) was used to observe the expression of CD86 and CD163, two important functional phenotypes of macrophage differentiation in lung tissue of UC rats, while the enzyme-linked immunosorbent assay (ELISA) was used to assess the content of IFN-γ, TNF-α, IL-4 and IL-13, the key cytokines of macrophage differentiation in micro environment of the lung tissue.

Results

Compared with the NG, the colons of rats in MG were injured more seriously, and the scores of gross observation and histological examination were significantly higher (P<0.05). Compared with the MG, the pathological changes of the two groups of rats with moxibustion treatment were improved, which presented with ulcer repair, inflammation dissipation, and the general score and histological score of the two groups were decreased (P<0.05). Compared with the NG, the expression of CD86 in the lung tissue of rats in the MG was increased (P<0.05), and CD163 expression was decreased (P<0.05). Compared with the MG and the SMG, the expression of CD86 in the lung tissue of the rats in the NMG was significantly lower than those in the MG and SMG, and CD163 was higher (P<0.05), while the differences were not statistically significant between the MG and the SMG (P>0.05). Compared with the NG, the expression of key cytokines in lung tissue of MG was abnormal, the contents of IFN-γ and TNF-α increased (P<0.05), while the IL-4 and IL-13 decreased (P<0.05). The IL-4 and IL-13 were significantly increased in lung tissue of rats in the NMG (P<0.05), and the IFN-γ and TNF-α were reduced (P<0.05), compared with those in the MG and the SMG, while the differences were not statistically significant between the MG and the SMG (P>0.05).

Conclusion

Moxibustion can increase the expression of CD163, the important functional phenotype of macrophages in lung tissue of UC rats, and the differentiation critical cytokines IL-4 and IL-13. It can also reduce activated phenotype CD86 and its differentiation critical cytokines IFN-γ and TNF-α in the lung tissue of UC rats.

摘要

目的

观察艾灸对溃疡性结肠炎(ulcerative colitis, UC)大鼠肺组织巨噬细胞分化重要功能表型CD86、CD163 表达的影响, 并基于巨噬细胞分化关键细胞因子干扰素γ (interferon-gamma, IFN-γ)、肿瘤坏死因子α(tumor necrosis factor-alpha, TNF-α)、白介素4 (interleukin-4, IL-4)和白介素13 (interleukin-13, IL-13)研究艾灸调节肺组织巨噬细胞分化的作用机制

方法

将40 只清洁级SD 大鼠随机分为正常组、模型组、普通艾灸组和无烟艾灸组, 采用抗原免疫结合局部福尔马林灌肠法制备UC 大鼠模型, 普通艾灸组大鼠双侧天枢穴接受艾灸治疗, 无烟艾灸组大鼠双侧天枢穴接受无烟艾灸治疗, 每次灸10 min, 每日1 次, 共8 d。实验结束, 采用苏木素-伊红 (hematoxylin-eosin, HE)染色法观察结肠组织病理学变化, 运用免疫印迹法(western blotting, WB)观察肺组织中巨 噬细胞分化重要功能表型CD86、CD163 的表达, 采用酶联免疫吸附测定法(enzyme-linked immunosorbent assay, ELISA)观察肺组织微环境中巨噬细胞分化关键细胞因子IFN-γ、TNF-α、IL-4、IL-13 的含量。

结果

与正常组比较, 模型组大鼠结肠组织损伤严重, 结肠大体评分及组织学评分明显升高(P<0.05)。与模型组比较, 接受艾灸治疗的 两组大鼠组织病变均有所改善, 表现为溃疡修复, 炎症减轻, 两组大鼠结肠大体评分及组织学评分均降低 (P<0.05)。与正常组比较, 模型组大鼠肺组织CD86 表达增加(P<0.05), CD163 表达降低(P<0.05)。与模型组、无 烟艾灸组比较, 普通艾灸组大鼠肺组织CD86 表达显著降低, CD163 表达升高(P<0.05), 而无烟艾灸组与模型组相 比, 差异无统计学意义(P>0.05)。与正常组比较, 模型组大鼠肺组织微环境中巨噬细胞分化关键细胞因子表达异 常, 表现为IFN-γ 和TNF-α 含量增加(P<0.05), IL-4 和IL-13 含量减少(均P<0.05); 与模型组及无烟艾灸组比较, 普 通艾灸组大鼠肺组织中IL-4、IL-13 含量明显升高, IFN-γ、TNF-α 含量减少(P<0.05), 无烟艾灸组与模型组相比, 差 异无统计学意义(P>0.05)。

结论

艾灸能上调UC 大鼠肺组织中巨噬细胞重要功能表型CD163 及分化关键细胞因 子IL-4、IL-13 表达, 下调活化表型CD86 及分化关键细胞因子IFN-γ、TNF-α 的表达。

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Authors and Affiliations

  1. Shanghai Research Institute of Acupuncture and Meridian, Shanghai, 200030, China

    Dan Zhang  (张丹), Jie Liu  (刘婕), Jue Hong  (洪珏), Zheng Shi  (施征), Huan-gan Wu  (吴焕淦) & Xiao-peng Ma  (马晓芃)

  2. Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China

    Dan Zhang  (张丹), Yan-ting Yang  (杨延婷), Jin-an Lin  (林晋安), Yi-fan Jia  (贾一凡), Yan Huang  (黄燕) & Zhi-yuan Li  (李志元)

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  1. Dan Zhang  (张丹)
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Correspondence to Xiao-peng Ma  (马晓芃).

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Zhang, D., Yang, Yt., Lin, Ja. et al. Regulatory effect of moxibustion for rats with ulcerative colitis on the macrophage functional phenotype protein of lung tissue. J. Acupunct. Tuina. Sci. 14, 1–9 (2016). https://doi.org/10.1007/s11726-016-0891-9

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  • DOI: https://doi.org/10.1007/s11726-016-0891-9

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