黄芪桂枝五物汤通过α7nAchR调节M1/M2巨噬细胞极化促进白色脂肪米色化

Huangqi Guizhi Wuwu Decoction promotes the beige of white adipose by regulating the polarization of M1/M2 macrophages throughα7nAchR

目的 观察黄芪桂枝五物汤对肥胖小鼠白色脂肪米色化作用的影响。方法 50只8周龄雄性C57BL/6J小鼠,随机选取其中10只作为正常饮食组(NF组),其余40只高脂饮食诱导小鼠肥胖。将诱导成功的40只肥胖小鼠随机分为高脂饮食组(HF组)、β3受体激动剂CL316,243组(β3组)、CL316,243+黄芪桂枝五物汤组(HGWD组)、CL316,243+黄芪桂枝五物汤+α7nAchR选择性拮抗剂α-BGT(α-BGT组),分别给予对应药物连续处理2周。HE染色观察白色脂肪细胞大小;PCR法检测白色脂肪组织中产热基因,M1、M2型巨噬细胞标志基因及α7nAchR基因水平;ELISA检测白色脂肪组织炎性因子含量。结果 与HF组比较,β3组体重下降,脂肪细胞面积减小,产热基因增加,表明β3受体激动剂促进了白色脂肪米色化;而M1、M2型巨噬细胞标志基因及抗炎因子含量未有显著变化,促炎因子含量增加,表明肥胖机体炎症状态未改善;同时α7nAchR基因水平下降。与β3组比较,HGWD组促白色脂肪米色化作用进一步增加,炎症状态显著改善,α7nAchR基因水平上升。而同时给予α-BGT后减弱了黄芪桂枝五物汤的上述作用。结论 黄芪桂枝五物汤可能通过上调白色脂肪组织α7nAchR水平,以抑制M1型巨噬细胞增加,促进M2型巨噬细胞极化,下调M1/M2型比值,减轻炎症反应,进而提高肥胖机体白色脂肪米色化效率。

Objective To observe the effect of Huangqi Guizhi Wuwu Decoction on the beige of white adipose in obese mice.Methods 10 of 508-week-old male C57BL/6J mice were randomly selected as normal diet group(NF group),and the other 40 mice were induced by high-fat diet.40 obese mice successfully induced were randomly divided into high-fat diet group(HF group),β3 receptor agonist CL316,243 group(β3 group),CL316,243+Huangqi Guizhi Wuwu Decoction group(HGWD group)and CL316,243+Huangqi Guizhi Wuwu Decoction+α7nAchR selective antagonistα-BGT group(α-BGT group),given corresponding drugs respectively for 2 weeks.HE staining to observe the area of white adipose.PCR to detect the levels of thermogenic gene,M1 and M2 macrophage marker gene andα7nAchR gene in white adipose tissue.ELISA to detect the contents of white adipose tissue inflammatory factors.Results Compared to HF group,the weight ofβ3 group decreased,the area of adi⁃pocytes decreased and the thermogenic genes increased,which indicated thatβ3 receptor agonist promoted the beige of white adi⁃pose;however,the contents of marker genes of M1 and M2 macrophages and anti-inflammatory factors have not changed signifi⁃cantly,but the contents of pro-inflammatory factors have increased,indicating that the inflammatory state of obese body has not relieved;and the level ofα7nAchR gene decreased.Compared toβ3 group,HGWD group further increased the beige of white ad⁃ipose,significantly relieved the inflammatory state and increased the level ofα7nAchR gene.However,the benefit effects of Huangqi Guizhi Wuwu decoction were weakened when co-administered withα-BGT.Conclusion Huangqi Guizhi Wuwu De⁃coction may inhibit the increase of M1 macrophages,promote the polarization of M2 macrophages,reduce the ratio of M1/M2,and then inhibit the inflammatory reaction,to improve the beige efficiency of white adipose in obese body.