丹龙醒脑方对血管性痴呆大鼠海马组织小胶质细胞活化及HMGB1/RAGE/NF-κB通路的影响  

Effects of Danlong Xingnao Prescription on the Activation of Microglial Cells in Hippocampal Region and HMGB1/RAGE/NF-κB Pathway in Vascular Dementia Model Rats

作　　者：张运辉 杨梦琳 周小青[2] 伍大华[2,3] 刘霞 秦建设[1] 黄玉静[1] ZHANG Yunhui;YANG Menglin;ZHOU Xiaoqing;WU Dahua;LIU Xia;QIN Jianshe;HUANG Yujing(Chongqing Three Gorges Medical College,Chongqing 404120,China;Hunan University of Chinese Medicine,Changsha 410208,China;Affiliated Hospital of Hunan Provincial Academy of Traditional Chinese Medicine,Changsha 410006,China)

机构地区：[1]重庆三峡医药高等专科学校,重庆404120 [2]湖南中医药大学,湖南长沙410208 [3]湖南省中医药研究院附属医院,湖南长沙410006

出　　处：《中国中医药信息杂志》2025年第4期120-127,共8页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81874462);重庆市教委科学技术研究计划项目(KJQN202302708);重庆市中医药传承创新团队(三峡库区道地药材保护与利用多学科交叉创新团队)(2022年);重庆市中医药重点学科(中医基础理论)建设项目(2021年);重庆市乡村振兴对口帮扶科技项目(2023TIAD-ZXX0010);重庆三峡医药高等专科学校校级创新团队-中医药防治呼吸和老年病团队(TIG202302);万州区博士直通车项目(wzst20230415);重庆三峡医药高等专科学校自然科学校级重点项目(2019XZZ005)。

摘　　要：目的基于HMGB1/RAGE/NF-κB通路探讨丹龙醒脑方对血管性痴呆(VD)大鼠学习记忆能力及小胶质细胞活化的影响。方法72只大鼠随机选取10只作为假手术组,其余大鼠采用改良双侧颈动脉结扎法制备VD大鼠模型。将造模成功的50只大鼠随机分为模型组、尼莫地平组(10.8 mg/kg)和丹龙醒脑方低、中、高剂量组(1.7、7.4、14.8 g/kg),每组10只,药物组分别予相应药液灌胃,假手术组和模型组灌胃等量生理盐水,连续28 d。Morris水迷宫实验评估大鼠学习记忆能力,HE染色观察海马组织形态,ELISA检测海马组织白细胞介素(IL)-1β、IL-6、肿瘤坏死因子-α(TNF-α)含量,RT-PCR检测海马组织高迁移率族蛋白B1(HMGB1)、晚期糖基化终末产物受体(RAGE)、核因子(NF)-κB p65、调节性RNA酶-1(Regnase-1)mRNA表达,免疫组化和Western blot检测海马组织离子钙结合适配分子1(Iba1)、HMGB1、RAGE、NF-κB p65、Regnase-1蛋白表达。结果与假手术组比较,模型组大鼠Morris水迷宫实验逃避潜伏期延长、穿越原平台次数增加(P<0.01),海马组织锥体细胞减少、形态不规则,细胞膜、核膜不清晰,可见较多坏死神经元,海马组织IL-1β、IL-6、TNF-α含量升高(P<0.01),海马组织HMGB1、RAGE、NF-κB p65 mRNA表达升高(P<0.01),Regnase-1 mRNA表达降低(P<0.01),Iba1、HMGB1、RAGE、NF-κB p65蛋白表达升高(P<0.01),Regnase-1蛋白表达降低(P<0.01);与模型组比较,丹龙醒脑方各剂量组大鼠逃避潜伏期缩短、穿越原平台次数减少(P<0.05,P<0.01),海马组织神经元结构明显改善,坏死神经元减少,海马组织IL-1β、IL-6、TNF-α含量降低(P<0.05,P<0.01),海马组织HMGB1、RAGE、NF-κB p65 mRNA表达降低,Regnase-1 mRNA表达升高(P<0.05,P<0.01),Iba1、HMGB1、RAGE、NF-κB p65蛋白表达降低,Regnase-1蛋白表达升高(P<0.05,P<0.01)。结论丹龙醒脑方能改善VD大鼠学习记忆能力,其机制可能与抑制HMGB1/RAGE/NF-κB通路激活及提高Regnase-1表达,进而�Objective To investigate the effects of Danlong Xingnao Prescription on learning and memory ability and microglia activation in rats with vascular dementia(VD)based on HMGB1/RAGE/NF-κB pathway.Methods Ten rats were randomly selected from 72 rats as a sham-operation group.The remaining rats were treated with modified bilateral common carotid artery ligation method to prepare the VD model.The 50 successful model rats were randomly divided into model group,nimodipine group(10.8 mg/kg)and Danlong Xingnao Prescription low-,medium-and high-dosage groups(3.7,7.4,14.8 g/kg),with 10 rats in each group.The administration groups were given relevant solution for gavage,the sham-operation group and model group were given the same amount of normal saline for consecutive 28 d.Morris water maze test was performed to evaluate learning and memory abilities of rats,the morphology in the hippocampus were observed by HE staining,the contents of interleukin(IL)-1β,IL-6 and tumor necrosis factor(TNF)-αin hippocampal tissue were detect by ELISA,RT-PCR was used to detect high mobility group protein B1(HMGB1),receptor of advanced glycation end product(RAGE),nuclear factor(NF)-κB p65 and regulatory RNase-1(Regnase-1)mRNA expression in hippocampal tissue,immunohistochemistry and Western blot were used to detect the protein expressions of ion calcium binding adapter molecule 1(Iba1),HMGB1,RAGE,NF-κB p65 and Regnase-1 in hippocampal tissue.Results Compared with the sham-operation group,the escape latency of rats was prolonged,and the number of crossings through the original platform was increased in the model group(P<0.01),the pyramidal cells in the hippocampus were reduced and irregularly shaped,with unclear cell and nuclear membranes,and a significant number of necrotic neurons were visible,the contents of IL-1β,IL-6 and TNF-αin hippocampal tissue increased(P<0.01),the mRNA expressions of HMGB1,RAGE and NF-κB p65 in hippocampal tissue increased(P<0.01),while the mRNA expression of Regnase-1 decreased(P<0.01),the protein expressions

关 键 词：丹龙醒脑方 血管性痴呆 学习记忆 HMGB1/RAGE/NF-κB通路 小胶质细胞 大鼠 

分 类 号：R285.5[医药卫生—中药学]