黑逍遥散调控SIRT1/p53/SLC7A11信号通路抑制铁死亡改善阿尔茨海默病模型大鼠认知功能障碍  

Hei Xiaoyaosan Inhibits Ferroptosis by Regulating SIRT1/p53/SLC7A11 Signaling Pathway to Ameliorate Cognitive Dysfunction in Rat Model of Alzheimer's Disease

作　　者：杨娇 陈怡琴 裴文丽 韩玉梅 王虎平[1,2,3] YANG Jiao;CHEN Yiqin;PEI Wenli;HAN Yumei;WANG Huping(Gansu University of Chinese Medicine,Lanzhou 730000,China;Gansu Key Laboratory of Traditional Chinese Medicine(TCM)Excavation and Innovative Transformation,Lanzhou 730000,China;Gansu Engineering Laboratory for New Products of TCM,Lanzhou 730000,China)

机构地区：[1]甘肃中医药大学,兰州730000 [2]甘肃省中医方药挖掘与创新转化重点实验室,兰州730000 [3]甘肃省中药新产品创制工程实验室,兰州730000

出　　处：《中国实验方剂学杂志》2025年第9期116-123,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82160862,81960828);甘肃省自然科学基金项目(22JR11RA113);第五批全国中医临床优秀人才研修项目(国中医药人教函〔2022〕239号);首批陇原青年英才项目(中共甘肃省委人才工作领导小组〔2022〕5号);甘肃省高校教师创新基金项目(2024A-083)。

摘　　要：目的:探讨黑逍遥散对阿尔茨海默病(AD)模型大鼠认知障碍及组蛋白脱乙酰酶sirtuin-1(SIRT1)/肿瘤抑制因子p53/溶质载体家族7A11(SLC7A11)信号通路的影响。方法:16周龄SPF级SD雄性大鼠90只,随机选取10只作为空白组,10只作为假手术组(双侧海马注射生理盐水1μL),其余70只双侧海马注射β淀粉样蛋白1-42(Aβ_(1-42))溶液1μL制备AD模型。按随机数字表法将造模成功的50只大鼠分为模型组,盐酸多奈哌齐组(0.45 mg·kg^(-1))及黑逍遥散高、中、低剂量组(15.30、7.65、3.82 g·kg^(-1)),连续灌胃42 d,1次/d。Morris水迷宫实验测试大鼠认知功能,尼色染色观察各组大鼠海马神经元的形态,普鲁士蓝染色检测海马组织铁沉积情况,生化试剂盒检测海马组织中超氧化物歧化酶(SOD)、丙二醛(MDA)、铁离子(Fe^(2+))含量,蛋白免疫印迹法(Western blot)及实时荧光定量聚合酶链式反应(Real-time PCR)检测大鼠海马组织SIRT1、p53、SLC7A11、谷胱甘肽过氧化物酶4(GPX4)、酰基辅酶A合成酶长链家族成员4(ACSL4)蛋白及mRNA的表达。结果:与空白组比较,模型组目标象限运动距离显著减少(P<0.01),目标象限滞留时间明显缩短(P<0.05),海马神经元细胞排列紊乱,海马区铁死亡沉积增多,SOD活性下降,MDA、Fe^(2+)含量明显上升(P<0.05,P<0.01),SIRT1、SLC7A11、GPX4蛋白表达及mRNA表达明显减少(P<0.05,P<0.01),p53、ACSL4蛋白及mRNA表达显著增加(P<0.01),AD病理进程加重;与模型组比较,盐酸多奈哌齐组目标象限滞留时间延长且目标象限运动距离明显增加(P<0.05,P<0.01),黑逍遥散高、中剂量组目标象限滞留时间延长且目标象限运动距离明显增加(P<0.05,P<0.01),神经元细胞状明显改善,海马区铁死亡沉积减少,SOD活性上升,MDA、Fe^(2+)含量均明显下降(P<0.05,P<0.01),SIRT1、SLC7A11、GPX4蛋白及mRNA表达明显增加(P<0.05,P<0.01),p53、ACSL4蛋白及mRNA表达明显减少(P<0.05,P<0.01)。结论:黑逍遥散可�Objective:To investigate the effects of Hei Xiaoyaosan on cognitive impairment and the histone deacetylase sirtuin-1(SIRT1)/tumor suppressor p53/solute carrier family 7 member 11(SLC7A11)signaling pathway in the rat model of Alzheimer's disease(AD).Methods:A total of 9016-week-old SPF-grade SD male rats were randomly assigned in a blank group(n=10),a sham group(n=10,with injection of 1μL normal saline into the bilateral hippocampi),and an AD modeling group(n=70,with injection of 1μLβamyloid 1-42 solution into the bilateral hippocampi).According to the random number table method,fifty successfully modeled rats were assigned into model,donepezil hydrochloride(0.45 mg·kg^(-1)),and high-,medium-,and lowdose(15.30,7.65,and 3.82 g·kg^(-1),respectively)Hei Xiaoyaosan groups,and they were administrated with corresponding agents via gavage once a day for 42 consecutive days.Morris water maze test was carried out to examine the cognitive function of rats.Nissl staining was employed to observe the morphology of hippocampal neurons in each group,and Prussian blue staining was used to detect iron deposition in the hippocampal tissue.Biochemical kits were used to measure the levels of superoxide dismutase(SOD),malondialdehyde(MDA),and iron ion(Fe^(2+))in the hippocampal tissue.Western blot and real-time quantitative polymerase chain reaction(Real-time PCR)were employed to determine the protein and mRNA levels,respectively,of SIRT1,p53,SLC7A11,glutathione peroxidase 4(GPX4),and acyl-CoA synthetase long-chain family member 4(ACSL4)in the hippocampus.Results:Compared with the blank group,the model group showed reductions in target quadrant movement distance(P<0.01)and target quadrant residence time(P<0.05),disarrangement of hippocampal neurons,increased ferroptosis deposition in the hippocampus,a lowered level of SOD,risen levels of MDA and Fe^(2+)(P<0.05,P<0.01),down-regulated protein and mRNA levels of SIRT1,SLC7A11,and GPX4(P<0.05,P<0.01),up-regulated protein and mRNA levels of p53 and ACSL4(P<0.01),and aggravated patholo

关 键 词：阿尔茨海默病 黑逍遥散 组蛋白脱乙酰酶sirtuin-1(SIRT1)/肿瘤抑制因子p53/溶质载体家族7A11(SLC7A11)信号通路 铁死亡 

分 类 号：R259[医药卫生—中西医结合] R285[医药卫生—中医内科学] R742[医药卫生—中医学]