摘要
目的:探讨黄连解毒汤通过调控F盒富含亮氨酸重复序列蛋白5(FBXL5)/铁调节蛋白2(IRP2)通路对血管性痴呆(VaD)小鼠脑铁代谢紊乱、髓鞘损伤及攻击行为的影响。方法:将60只C57BL/6J小鼠随机分为假手术组、模型组、利培酮组(2 mg·kg^(-1)·d^(-1))和黄连解毒汤低、中、高剂量组(0.25、0.5、1 g·kg^(-1)·d^(-1)),每组10只。采用双侧颈总动脉狭窄(BCAS)法建立VaD模型。从第7周开始给予药物干预2周。在第9周进行行为学评估,包括触摸逃避实验和居民入侵者测试。行为学测试结束后,收集下丘脑腹外侧腹内侧(VMHvl)组织样本。蛋白免疫印迹法(Western blot)检测髓鞘相关糖蛋白(MAG)、髓鞘少突胶质细胞糖蛋白(MOG)、髓鞘碱性蛋白(MBP)、4-羟基-2-壬烯醛(4-HNE)、谷胱甘肽过氧化物酶4(GPX4)、转铁蛋白受体1(TFR1)、铁蛋白轻链(Ft-L)、膜铁转运蛋白1(FPN1)、FBXL5和IRP2表达;免疫荧光检测MBP荧光强度;透射电镜观察髓鞘超微结构改变;Perl's染色检测组织铁沉积;酶联免疫吸附测定法(ELISA)检测丙二醛(MDA)和超氧化物歧化酶(SOD)水平。结果:与假手术组比较,模型组小鼠啃咬次数、攻击行为、易激惹评分明显升高,攻击潜伏期显著缩短(P<0.01);MAG、MOG和MBP的表达水平及MBP的荧光强度均显著降低(P<0.01),髓鞘超微结构紊乱。Ft-L、TFR1表达升高,FPN1表达降低,铁沉积增加(P<0.01)。抗氧化剂GPX4和SOD降低,4-HNE和脂质过氧化产物MDA升高(P<0.01)。FBXL5蛋白表达降低,IRP2蛋白表达升高(P<0.01)。与模型组比较,黄连解毒汤中、高剂量组和利培酮组小鼠啃咬次数、攻击行为和激惹评分降低(P<0.05,P<0.01),攻击潜伏期增加(P<0.01);黄连解毒汤中、高剂量组MAG、MOG、MBP表达水平及MBP免疫荧光均升高(P<0.01)。超微板层结构排列有序;Ft-L、TFR1表达降低,FPN1表达升高,铁沉积减少(P<0.01);GPX4、SOD表达水平升高,4-HNE、MDA表达水平降低(P<0.01);FBXL5蛋白�
Objective:This study aims to investigate the effect of Huanglian Jiedutang on brain iron metabolism disorders,myelin damage,and aggressive behavior in vascular dementia(VaD)mice by regulating the F-box leucine-rich repeat protein 5(FBXL5)/iron regulatory protein 2(IRP2)pathway.Method:Sixty C57BL/6J mice were randomly divided into six groups:sham operation group,model group,risperidone group(2 mg·kg^(-1)·d^(-1)),and low-dose,medium-dose,and high-dose groups of Huanglian Jiedutang(0.25,0.5,1 g·kg^(-1)·d^(-1)),with 10 mice in each group.The VaD model was established by bilateral carotid artery stenosis(BCAS).Drug intervention was administered for two weeks starting from the seventh week.Behavioral assessments,including the touch escape and resident-intruder tests,were conducted in the ninth week.After the behavioral tests,ventromedial hypothalamus ventrolateral(VMHvl)tissue samples were collected.Western blot was used to detect the expression of myelin-associated glycoprotein(MAG),myelin oligodendrocyte glycoprotein(MOG),myelin basic protein(MBP),4-hydroxynonenal(4-HNE),glutathione peroxidase 4(GPX4),transferrin receptor 1(TFR1),ferritin light chain(Ft-L),ferroportin 1(FPN1),FBXL5,and IRP2.Immunofluorescence was used to measure MBP fluorescence intensity.Transmission electron microscopy was employed to observe ultrastructural changes in the myelin sheath.Perl's staining was used to detect tissue iron deposition.Malondialdehyde(MDA)and superoxide dismutase(SOD)levels were measured by enzyme linked immunosorbent assay(ELISA).Result:Compared to the sham operation group,the model group exhibited a significant increase in biting,aggression,and irritability scores,along with a reduced latency of the attack(P<0.01).The expression levels of MAG,MOG,and MBP,as well as the fluorescent density of MBP,were significantly decreased(P<0.01).Disordered myelin ultrastructure,increased Ft-L and TFR1 expression,decreased FPN1 expression,and elevated iron deposition were observed(P<0.01).Antioxidants GPX4 and SOD were reduced,while
作者
冯小霞
陈晨
邱静
阳光
FENG Xiaoxia;CHEN Chen;QIU Jing;YANG Guang(The First Clinical Medical School,Hubei University of Chinese Medicine,Wuhan 430061,China;Hubei Provincial Hospital of Traditional Chinese Medicine(TCM),Wuhan 430060,China;Hubei Provincial Institute of TCM,Hubei Key Laboratory of Theory and Application Research of Liver and Kidney in TCM,Wuhan 430060,China;Hubei Shizhen Laboratory,Wuhan 430060,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2024年第24期1-9,共9页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金面上项目(82274334)
国家自然科学基金青年项目(82205084)
武汉市应用基础前沿项目(2020020601012244)。
关键词
黄连解毒汤
铁代谢紊乱
髓鞘损伤
血管性痴呆
激越攻击行为
Huanglian Jiedutang
iron metabolism disorder
myelin damage
vascular dementia
aggressivebehavior
