艾灸通过白细胞介素-33/生长刺激表达基因2蛋白通路影响阿尔茨海默病小鼠海马小胶质细胞极化  

M oxibustion influences hippocampal microglia polarization via IL-33/ST2 pathway in Alzheimer’s disease mice

作　　者：李虹莹 沈缘 谢璐霜[2] 吴巧凤[1] 余曙光[1] LI Hong-ying;SHEN Yuan;XIE Lu-shuang;WU Qiao-feng;YU Shu-guang(Acupuncture-moxibustion and Tuina College,Chengdu University of Traditional Chinese Medicine,Chengdu 610075,China;College of Basic Medicine,Chengdu University of Traditional Chinese Medicine,Chengdu 610075)

机构地区：[1]成都中医药大学针灸推拿学院,成都610075 [2]成都中医药大学基础医学院,成都610075

出　　处：《针刺研究》2023年第12期1202-1208,1235,共8页Acupuncture Research

基　　金：国家自然科学基金项目(No.81873386、82174511、82174512)。

摘　　要：目的:观察艾灸通过白细胞介素-33(IL-33)/生长刺激表达基因2蛋白(ST2)信号通路对阿尔茨海默病(AD)小鼠小胶质细胞向M2方向极化的影响。方法:5月龄APP/PS1雄性小鼠随机分为模型组和艾灸组,同月龄C57BL/6J小鼠为对照组,每组9只。艾灸组温和灸“百会”“涌泉”,每次30 min,每日1次,每周5 d,共4周。采用Morris水迷宫实验评估小鼠空间学习记忆能力,Western blot法检测小鼠海马组织IL-33和ST2蛋白表达量,免疫荧光染色法检测小鼠海马CA1区β-淀粉样蛋白(Aβ)、磷酸化Tau蛋白(p-Tau)阳性表达水平及IL-33/离子钙接头蛋白分子1(Iba-1)、ST2/Iba-1、精氨酸酶1(Arg1)/Iba-1和诱导型一氧化氮合酶(iNOS)/Iba-1的荧光双标共表达情况。结果:与对照组比较,模型组小鼠各时点逃避潜伏期延长(P<0.001,P<0.01),进入有效区域次数和目标象限游泳时间百分比下降(P<0.001),海马CA1区Aβ和p-Tau蛋白阳性表达水平升高(P<0.001),海马组织IL-33和ST2蛋白表达降低(P<0.05,P<0.001),海马CA1区IL-33/Iba-1、ST2/Iba-1、Arg1/Iba-1蛋白共表达水平降低(P<0.001),iNOS/Iba-1蛋白共表达水平升高(P<0.001)。与模型组比较,艾灸组小鼠各时点逃避潜伏期缩短(P<0.001,P<0.01),进入有效区域次数和在目标象限游泳时间百分比升高(P<0.001),海马CA1区Aβ和p-Tau蛋白阳性表达水平降低(P<0.001),海马组织IL-33和ST2蛋白表达升高(P<0.05,P<0.01),海马CA1区IL-33/Iba-1、ST2/Iba-1、Arg1/Iba-1蛋白共表达水平升高(P<0.001,P<0.05,P<0.01),iNOS/Iba-1蛋白共表达水平降低(P<0.001)。结论:艾灸可以改善APP/PS1小鼠的空间学习记忆能力,减少Aβ和p-Tau的病理沉积,可能与上调IL-33/ST2信号通路、促进小胶质细胞向M2方向极化有关。ObjectiveTo observe the effect of m oxibustion on the polarization of m icroglia towards M2 direc⁃tion in Alzheim er’s disease(AD)m ice through the interleukin-33(IL-33)/growth stim ulating gene 2 protein(ST2)sig⁃naling pathway.MethodsFive-m onth-old APP/PS1 m ale m ice were random ly divided into m odel and m oxibustion(M oxi)groups,and C57BL/6J m ice of the sam e age were as the control group,with 9 m ice in each group.In the M oxi group,m oxibustion was applied at“Baihui”(GV20)and“Yongquan”(KI1)for 30 m in,once a day,5 days a week for 4 weeks.The spatial learning m em ory ability was observed by the M orris water m aze test.The relative expressions of IL-33and ST2 in hippocam pus were detected by W estern blot.The positive expression of am yloid-β(Aβ),phosphorylated Tau(p-Tau),IL-33/ionized calcium binding adapter m olecule 1(Iba-1),ST2/Iba-1,arginase 1(Arg1)/Iba-1 and indu-cible nitric oxide synthase(iNOS)/Iba-1 in hippocam pal CA1 region were detected by im m unofluorescence.ResultsCom pared with the control group,the escape latency of the m ice in the m odel group was prolonged(P<0.001,P<0.01),the num ber of tim es to enter the effective area and the percentage of target quadrant swim m ing tim e were re⁃duced(P<0.001),the positive expression of both Aβand p-Tau,the positive expression of iNOS/Iba-1 in the hippocam⁃pal CA1 region was increased(P<0.001),while the expression of IL-33 and ST2 protein in hippocam pal tissue,the positive expression levels of IL-33/Iba-1,ST2/Iba-1 and Arg1/Iba-1 in hippocam pal CA1 region were all decreased(P<0.05,P<0.001).After treatm ent,com pared with the m odel group,the escape latency of the m ice in the m oxibustion group was shortened(P<0.001,P<0.01),the num ber of entries into the effective area and the percentage of target quadrant swim m ing tim e were increased(P<0.001),the positive expression of Aβand p-Tau in the hippocam pal CA1region,and the positive expression of iNOS/Iba-1 were decreased(P<0.001),while the expression of IL-33 and ST2protein

关 键 词：艾灸 阿尔茨海默病 IL-33/ST2信号通路 小胶质细胞极化 

分 类 号：R245.8[医药卫生—针灸推拿学]