穴位埋线对肥胖小鼠附睾脂肪组织巨噬细胞极化的影响

Effect of acupoint thread-embedding on macrophage polarization of epididymis adipose tissue in obese mice

目的:观察"足三里""丰隆"穴位埋线对肥胖小鼠附睾脂肪组织巨噬细胞极化的影响,探讨穴位埋线减重的作用机制。方法:从30只雄性C57BL/6小鼠中随机选取10只予普通饲料喂养,剩余20只予高脂饲料喂养制备肥胖模型。将造模成功的16只小鼠随机分为模型组和埋线组,每组8只,从普通饲料喂养的小鼠中随机选取8只作为正常组。造模成功次日,埋线组于"足三里""丰隆"穴行穴位埋线,每10天1次,共治疗4次。比较各组小鼠干预第0、8、16、24、32、40天的体质量及干预后糖代谢水平。干预结束后,比较各组小鼠脂代谢水平和附睾脂肪组织质量;HE染色法观察各组小鼠附睾脂肪组织形态;实时荧光定量PCR法检测小鼠附睾脂肪组织M1和M2型巨噬细胞相关细胞因子白介素-6(IL-6)、单核细胞趋化蛋白-1(MCP-1)、肿瘤坏死因子-α(TNF-α)、白介素-10(IL-10)的mRNA表达;实时荧光定量PCR法和Western blot法检测小鼠附睾脂肪组织M1型巨噬细胞表面标志物诱导型一氧化氮合酶(iNOS)和M2型巨噬细胞表面标志物精氨酸酶1(Arg-1)mRNA和蛋白的表达。结果:与正常组比较,模型组小鼠干预第0、8、16、24、32、40天体质量均增加(P<0.05);葡萄糖耐量实验0、30、60、120 min及胰岛素耐受实验0、30、60、90、120 min血糖均高于正常组(P<0.05);总胆固醇和三酰甘油水平明显高于正常组(P<0.001,P<0.01);附睾脂肪组织质量明显高于正常组(P<0.001);附睾脂肪组织IL-6、MCP-1、TNF-α及iNOS mRNA表达升高(P<0.05,P<0.01,P<0.001),IL-10、Arg-1 mRNA表达降低(P<0.01),iNOS蛋白表达上调(P<0.01),Arg-1蛋白表达下调(P<0.001)。与模型组比较,埋线组小鼠在干预16、24、32、40天体质量降低(P<0.05);葡萄糖耐量实验30、60、120 min及胰岛素耐受实验30、60 min血糖均低于模型组(P<0.05);总胆固醇和三酰甘油水平低于模型组(P<0.01,P<0.05);附睾脂肪组织质量明显低于模型组(P<0.01);附睾脂肪组织IL-6、MCP-1、TNF-α、iNOS mRNA表达降低(P<0.05),IL-10、Arg-1 mRNA表达升高(P<0.05),iNOS蛋白表达下调(P<0.05),Arg-1蛋白表达上调(P<0.01)。结论:"足三里""丰隆"穴位埋线可能通过影响巨噬细胞的极化而发挥减重作用。

Objective To observe the effect of acupoint thread-embedding at "Zusanli"(ST 36) and "Fenglong"(ST 40) on the macrophage polarization of epididymis adipose tissue in obese mice, and to explore the action mechanism of acupoint thread-embedding on weight control. Methods Among 30 male C57 BL/6 mice, 10 mice were randomly selected and fed with normal diet, and the remaining 20 mice were fed with high-fat diet to establish the obesity model. Sixteen mice with successful obesity model were randomly divided into a model group and an acupoint thread-embedding group, 8 mice in each group. Eight mice were selected from mice which were fed with normal diet as the normal group. On the next day of successful modeling, acupoint thread-embedding was performed at "Zusanli"(ST 36) and "Fenglong"(ST 40) in the acupoint thread-embedding group, once every 10 days for 4 times. The body weight was recorded at 0, 8, 16, 24, 32, 40 days into intervention;the level of glucose metabolism was compared after intervention;the level of lipid metabolism and weight of epididymal adipose tissue were compared at the end of the intervention;the mRNA expression of M1 and M2 macrophage-related cytokines interleukin-10(IL-6), monocyte chemoattractant protein-1(MCP-1), tumor necrosis factor-α(TNF-α) and interleukin-10(IL-10) were detected by real-time PCR;the mRNA and protein expression of M1 macrophage labeled inducible nitric oxide synthase(iNOS) and M2 macrophage labeled arginase-1(Arg-1) were detected by real-time PCR and Western blot. Results Compared with the normal group, the body weight at 0, 8, 16, 24, 32, 40 days into intervention in the model group was increased(P<0.05);the results of glucose tolerance test at 0, 30, 60, 120 min and insulin tolerance test at 0, 30, 60, 90, 120 min in the model group were higher than those in the normal group(P<0.05);the levels of total cholesterol and triacylglycerol in the model group were significantly higher than those in the normal group(P<0.001, P<0.01);the weight of epididymal adipose tissue in the model group was significantly higher than that in the normal group(P<0.001);the mRNA expression of IL-6, MCP-1, TNF-α and iNOS was increased(P<0.05,P<0.01, P<0.001), that of IL-10, Arg-1 was decreased(P<0.01), the protein expression of iNOS was up-regulated(P<0.01), and that of Arg-1 was down-regulated(P<0.001). Compared with the model group, the body weight at 16, 24, 32, 40 days into treatment in the acupoint thread-embedding group was reduced(P<0.05);the results of glucose tolerance test at 30, 60, 120 min and insulin tolerance test at 30, 60 min in the acupoint thread-embedding group were lower than those in the model group(P<0.05);the levels of total cholesterol and triacylglycerol in the acupoint thread-embedding group were significantly lower than those in the model group(P<0.01, P<0.05);the weight of epididymal adipose tissue in the acupoint thread-embedding group was significantly lower than that in the model group(P<0.01);the mRNA expression of IL-6, MCP-1, TNF-α and iNOS was reduced(P<0.05), that of IL-10, Arg-1 was increased(P<0.05), the protein expression of iNOS was down-regulated(P<0.05), and that of Arg-1 was up-regulated(P<0.01). Conclusion Acupoint thread-embedding at "Zusanli"(ST 36) and "Fenglong"(ST 40) may play a role in weight control by regulating the polarization of macrophages.