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[针刺对高脂饮食诱导的肥胖小鼠白色脂肪组织巨噬细胞极化的影响]

[Effect of acupuncture on macrophage polarization of white adipose tissue in obese mice induced by high-fat diet].

作者信息

Zhang Siyi, Hu Xia, Tang Hongtu

机构信息

College of Acupuncture and Orthopedics, Hubei University of CM, Wuhan 430061, China.

Basic Medical College, Hubei University of CM.

出版信息

Zhongguo Zhen Jiu. 2017 Nov 12;37(11):1205-11. doi: 10.13703/j.0255-2930.2017.11.018.

Abstract

OBJECTIVE

To observe the effect of acupuncture at acupoints and non-acupoints on macrophage polarization of white adipose tissue in obese mice induced by high-fat diet(HFD).

METHODS

Fifty male C57BL/6 mice were randomly divided into a control group (=10), and other mice were fed with HFD for 16 weeks to establish model. Thirty model mice, which were successfully established and continuously fed with HFD for 8 weeks, were randomized into a model group, a non-acupoint group and an acupuncture group (=10/group). The mice in the control group were continuously fed with common food for 8 weeks. From the second day after model established, the mice of the acupuncture group were treated with acupuncture at "Guanyuan" (CV 4), "Zusanli" (ST 36) and "Weiwanxiashu" (EX-B 3); the mice of the non-acupoint group were treated with acupuncture at two non-acupoints in tail, 15 min a time, once a day for 8 weeks. After model establishment and on the second day after the last acupuncture treatment, the body weight was recorded. The level of serum triglyceride (TG), total cholesterol (TC) in eye socket blood were measured. The morphological structure of epididymis white adipose tissue (eWAT) in mice was observed by hematoxylin and eosin (HE) staining. The mRNA expressions of interleukin-6 (IL-6), monocyte chemotactic protein 1 (MCP-1), tumor necrosis factor α (TNF-α), IL-10, inducible nitric oxide synthesis (iNOS), CD206 in eWAT of mice were determined with real-time quantitative PCR (RT-qPCR). The protein expressions of iNOS and CD206 in eWAT were determined with immunohistochemistry staining (IHC).

RESULTS

Compared with the control group, the body weight in the model, non-acupoint and acupuncture groups increased at the 16th and 24th weeks (all <0.05). Compared with the model group, the body weight of the acupuncture group at the 24th week decreased (<0.05). The serum TG and TC in the model group increased compared with those of the control group (both <0.05). The TC in the non-acupoint group decreased (<0.05), and the TG and TC in the acupuncture group decreased compared with those in the model group (both <0.05). Meanwhile, the mRNA expressions of MCP-1, TNF-α, IL-6 and iNOS in the model and non-acupoint groups increased compared with those in the control group (all <0.05), and the mRNA expressions of IL-10 and CD206 decreased (all <0.05). The mRNA expressions of MCP-1, TNF-α, IL-6 and iNOS in the acupuncture group were lower than those in the model group (all <0.05), and the mRNA expressions of IL-10 and CD206 were higher (both <0.05). HE staining showed alveolate adipose tissue in the control group with numbers of unilocular adipose cells, vacuolar polygon or circle. The adipose cells in the model group were larger and irregular with larger intercellular space. The adipose cells in the acupuncture group were minified with smaller intercellular space. Compared with those in the control group, the protein expressions of iNOS increased (both <0.05) and those of CD206 decreased (both <0.05) in the model and non-acupoint groups. Compared with those in the model group, the protein expression of iNOS decreased (<0.05) and that of CD206 increased (<0.05) in the acupuncture group.

CONCLUSION

Acupuncture can improve inflammation of obese mice through affecting macrophage polarization of white adipose tissue.

摘要

目的

观察针刺穴位与非穴位对高脂饮食(HFD)诱导的肥胖小鼠白色脂肪组织巨噬细胞极化的影响。

方法

将50只雄性C57BL/6小鼠随机分为对照组(n = 10),其余小鼠给予HFD喂养16周以建立模型。将30只成功建立模型并继续给予HFD喂养8周的小鼠随机分为模型组、非穴位组和针刺组(每组n = 10)。对照组小鼠连续给予普通饲料8周。从造模后第二天起,针刺组小鼠针刺“关元”(CV 4)、“足三里”(ST 36)和“胃脘下俞”(EX - B 3);非穴位组小鼠针刺尾部两个非穴位,每次15分钟,每天1次,共8周。造模后及末次针刺治疗后第二天记录体重。测量眼眶血中血清甘油三酯(TG)、总胆固醇(TC)水平。采用苏木精-伊红(HE)染色观察小鼠附睾白色脂肪组织(eWAT)的形态结构。用实时定量PCR(RT - qPCR)检测小鼠eWAT中白细胞介素 - 6(IL - 6)、单核细胞趋化蛋白1(MCP - 1)、肿瘤坏死因子α(TNF - α)、IL - 10、诱导型一氧化氮合酶(iNOS)、CD206的mRNA表达。用免疫组织化学染色(IHC)检测eWAT中iNOS和CD206的蛋白表达。

结果

与对照组相比,模型组、非穴位组和针刺组在第16周和第24周体重均增加(均P < 0.05)。与模型组相比,针刺组在第24周体重下降(P < 0.05)。模型组血清TG和TC较对照组升高(均P < 0.05)。非穴位组TC下降(P < 0.05),针刺组TG和TC较模型组下降(均P < 0.05)。同时,模型组和非穴位组中MCP - 1、TNF - α、IL - 6和iNOS的mRNA表达较对照组升高(均P < 0.05),IL - 10和CD206的mRNA表达下降(均P < 0.05)。针刺组中MCP - 1、TNF - α、IL - 6和iNOS的mRNA表达低于模型组(均P < 0.05),IL - 10和CD206的mRNA表达较高(均P < 0.05)。HE染色显示对照组为蜂窝状脂肪组织,有许多单泡脂肪细胞,呈空泡状多边形或圆形。模型组脂肪细胞较大且不规则,细胞间隙较大。针刺组脂肪细胞缩小,细胞间隙较小。与对照组相比,模型组和非穴位组中iNOS的蛋白表达增加(均P < 0.05),CD206的蛋白表达下降(均P < 0.05)。与模型组相比,针刺组中iNOS的蛋白表达下降(P < 0.05),CD206的蛋白表达增加(P < 0.05)。

结论

针刺可通过影响白色脂肪组织巨噬细胞极化改善肥胖小鼠的炎症反应。

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