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

目的:观察穴位与非穴位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。方法:将50只雄性C 57 BL/6小鼠随机抽取10只作为正常组,其余40只给予持续高脂饮食造模16周,将30只造模成功的小鼠随机分为模型组、非穴位组、穴位组,每组10只,继续给予高脂饮食喂养8周。正常组继续给予普通饲料喂养8周。自造模成功后第2天开始,穴位组小鼠针刺"关元""足三里""胃脘下俞",非穴位组针刺尾部2个非穴位,每日1次,每次针刺15 min,连续治疗8周。于造模成功后及末次治疗后第2天记录各组小鼠体质量;眼眶采血,检测小鼠血清三酰甘油(triglyceride,TG)、血清总胆固醇(total cholesterol,TC)水平;HE染色观察各组小鼠附睾白色脂肪组织形态,real-time quantitative PCR(RT-q PCR)检测小鼠附睾白色脂肪中白细胞介素6(interleukin-6,IL-6)、单核细胞趋化蛋白-1(monocyte chemotactic protein 1,MCP-1)、肿瘤坏死因子α(tumor necrosis factorα,TNF-α)、白细胞介素10(interleukin-10,IL-10)、诱导型一氧化氮合酶(inducible nitric oxide synthesis,i NOS)和CD 206 m RNA表达情况,免疫组化(IHC)检测附睾白色脂肪组织中i NOS和CD 206的蛋白表达情况。结果:与正常组比较,模型组、非穴位组、穴位组小鼠在喂养第16周和第24周时体质量均明显上升(均P<0.05);与模型组比较,穴位组小鼠在第24周时体质量明显降低(P<0.05)。与正常组比较,模型组TG、TC明显上升(均P<0.05);与模型组比较,非穴位组TC明显降低(P<0.05),穴位组TG、TC明显降低(均P<0.05)。与正常组比较,模型组和非穴位组小鼠脂肪组织中MCP-1、TNF-α、IL-6、i NOS m RNA的表达明显升高(均P<0.05),IL-10、CD 206 m RNA的表达明显降低(均P<0.05);与模型组比较,穴位组脂肪组织中MCP-1、TNF-α、IL-6、i NOS m RNA的表达明显降低(均P<0.05),IL-10、CD 206 m RNA的表达明显升高(均P<0.05)。HE染色可见:正常组脂肪组织呈蜂窝状结构,由大量单泡脂肪细胞构成,脂肪细胞呈多边形或圆形,呈空泡状;模型组脂肪细胞明显增大且脂肪细胞大小不规则,细胞间隙变大;穴位组脂肪细胞变小,细胞间隙变小。与正常组比较,模型组和非穴位组小鼠脂肪组织中i NOS蛋白表达明显升高(均P<0.05),CD206蛋白表达明显降低(均P<0.05);与模型组比较,穴位组脂肪组织中i NOS蛋白表达明显降低(P<0.05),CD206蛋白表达明显升高(P<0.05)。结论:针刺"关元""足三里""胃脘下俞"穴可通过影响白色脂肪组织巨噬细胞极化改善肥胖小鼠脂肪炎性反应。

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 C57 BL/6 mice were randomly divided into a control group（n=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（n=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（e WAT） in mice was observed by hematoxylin and eosin（HE） staining. The m RNA expressions of interleukin-6（IL-6）, monocyte chemotactic protein 1（MCP-1）, tumor necrosis factor α（TNF-α）, IL-10,inducible nitric oxide synthesis（i NOS）, CD206 in e WAT of mice were determined with real-time quantitative PCR（RT-q PCR）. The protein expressions of i NOS and CD206 in e WAT 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 16 th and 24 th weeks（all P0.05）. Compared with the model group, the body weight of the acupuncture group at the 24 th week decreased（P0.05）. The serum TG and TC in the model group increased compared with those of the control group（both P0.05）. The TC in the non-acupoint group decreased（P0.05）, and the TG and TC in the acupuncture group decreased compared with those in the model group（both P0.05）. Meanwhile, the m RNA expressions of MCP-1, TNF-α, IL-6 and i NOS in the model and non-acupoint groups increased compared with those in the control group（all P0.05）, and the m RNA expressions of IL-10 and CD206 decreased（all P0.05）. The m RNA expressions of MCP-1,TNF-α, IL-6 and i NOS in the acupuncture group were lower than those in the model group（all P0.05）, and the m RNA expressions of IL-10 and CD206 were higher（both P0.05）. 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 i NOS increased（both P0.05） and those of CD206 decreased（both P0.05） in the model and non-acupoint groups. Compared with those in the model group, the protein expression of i NOS decreased（P0.05） and that of CD206 increased（P0.05） in the acupuncture group. Conclusion Acupuncture can improve inflammation of obese mice through affecting macrophage polarization of white adipose tissue.