基于海马Mst1/NF-κB p65通路探讨化瘀通络灸对血管性痴呆大鼠学习记忆能力的影响  

Effect of Huayu Tongluo moxibustion on learning-memory ability in rats with vascular dementia based on hippocampal Mst1/NF-κB p65 pathway

作　　者：王萍 杨骏[3] 孔钰[4] 张雅婷[5] 樊吟秋 石海平 刘兰英 WANG Ping;YANG Jun;KONG Yu;ZHANG Yating;FAN Yinqiu;SHI Haiping;LIU Lanying(Afiliated Hospital of Nanjing University of Chinese Medicine/Jiangsu Provincial Hospital of TCM,Nanjing 210029,China;Key Laboratory of Acupuncture and Medicine Research of Ministry of Education,Nanjing University of Chinese Medicine,Nanjing 210023,Jiangsu Province;First Afiliated Hospital of Anhui University of CM,Hefei 230031;Shenzhen Baoan District Hospital of TCM;Anhui University of CM)

机构地区：[1]南京中医药大学附属医院/江苏省中医院,南京210029 [2]南京中医药大学针药结合教育部重点实验室,南京210023 [3]安徽中医药大学第一附属医院,合肥230031 [4]深圳市宝安区中医院 [5]安徽中医药大学

出　　处：《中国针灸》2025年第1期53-60,共8页Chinese Acupuncture & Moxibustion

基　　金：国家自然科学基金资助项目:82104994、82274632;安徽省自然科学基金项目:109066010008;南京中医药大学自然科学基金项目:XZR2020007。

摘　　要：目的:观察化瘀通络灸对血管性痴呆(VD)大鼠学习记忆能力及海马炎性反应相关通路哺乳动物不育系20样激酶1(Mst1)/核转录因子κB(NF-κB)p65的影响。方法:将60只SPF级雄性Wistar大鼠随机分为假手术组(12只)和造模组(48只),造模组大鼠采用改良双侧颈总动脉永久结扎方法建立VD模型,将造模成功的36只大鼠随机分为模型组、艾灸组和西药组,每组12只。艾灸组大鼠予化瘀通络灸,穴取“大椎”“百会”“神庭”,每次20 min,每日1次,7 d为一疗程,疗程间休息1 d,共干预3个疗程。西药组予0.72 mg/kg吡拉西坦灌胃,每天2次,疗程与艾灸组相同。应用Morris水迷宫实验检测各组大鼠学习记忆能力,HE染色法观察大鼠海马CA1区形态,实时荧光定量PCR法检测大鼠海马Mst1、M1型小胶质细胞标志物CD86、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)mRNA表达,ELISA法检测大鼠海马IL-6、TNF-α含量,Westernblot法检测大鼠海马Mst1、NF-κB p65蛋白表达。结果:与假手术组比较,模型组大鼠逃避潜伏期延长(P<0.05);与模型组比较,艾灸组和西药组大鼠逃避潜伏期均缩短(P<0.05)。模型组大鼠海马CA1区细胞排列无序,细胞塌陷,细胞核不规则;与模型组比较,艾灸组和西药组大鼠海马CA1区细胞排列较规整,损伤程度好转。与假手术组比较,模型组大鼠海马Mst1、CD86、IL-6、TNF-αm RNA及Mst1、NF-κB p65蛋白表达升高(P<0.05);与模型组比较,艾灸组和西药组大鼠海马Mst1、CD86、IL-6、TNF-αm RNA及Mst1、NF-κBp65蛋白表达降低(P<0.05)。与假手术组比较,模型组大鼠海马IL-6、TNF-α含量升高(P<0.05);与模型组比较,艾灸组和西药组大鼠海马IL-6、TNF-α含量降低(P<0.05)。结论:化瘀通络灸可改善VD大鼠学习记忆能力,作用机制可能为通过Mst1/NF-κB p65通路调控小胶质细胞激活,减少促炎因子IL-6、TNF-α的释放,从而缓解炎性因子对VD大鼠海马的损害。Objective To observe the effects of Huayu Tongluo(transforming stasis and unblocking collaterals)moxibustion on learning-memory ability and hippocampal mammalian sterile 20-like kinase 1(Mst1)/nuclear factorκB(NF-κB)p65 pathway related to inflammatory response in rats with vascular dementia(VD).Methods A total of 60 male Wistar rats of SPF grade were randomly divided into a sham operation group(12 rats)and a modeling group(48 rats).VD model was established by the method of modified bilateral common carotid artery permanent ligation in the modeling group.Thirty-six rats with successful modeling were randomly divided into a model group,a moxibustion group and a western medication group,with 12 rats in each group.Huayu Tongluo moxibustion was applied at"Dazhui"(GV14),"Baihui"(GV20)and"Shenting"(GV24)in the moxibustion group,20 min each time,once a day,7 day-intervention was as one course,and 1 day-interval was taken between two courses,for a total of 3 courses.In the western medication group,piracetam was given 0.72 mg/kg by intragastric administration,twice a day,the course of intervention was same as that of the moxibustion group.The learning-memory ability was detected by Morris water maze test;the morphology of hippocampal CA1 region was observed by HE staining;the mRNA expression of Mst1,M1 microglia markers CD86,interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)was detected by real-time PCR;the levels of IL-6 and TNF-αin hippocampus were detected by ELISA;and the protein expression of Mst1 and NF-κB p65 in hippocampus was detected by Western blot in rats of each group.Results Compared with the sham operation group,the escape latency was prolonged in the model group(P<0.05);compared with the model group,the escape latency was shortened in the moxibustion group and the western medication group(P<0.05).The cells in the CA1 region of hippocampus were disordered,cell collapse and irregular nuclei could be observed in the model group;compared with the model group,the cell arrangement in the CA1 region of hi

关 键 词：血管性痴呆 化瘀通络灸 海马 Mst1/NF-κB p65 神经炎性反应 小胶质细胞 

分 类 号：R245[医药卫生—针灸推拿学]