异功散调控CXCL12/CXCR4信号减少谷氨酸释放改善衰老模型小鼠认知下降  

Yigong Powder regulates CXCL12/CXCR4 signaling to reduce glutamate release and prevent cognitive decline in mouse model of aging

作　　者：魏江平 赵子瑄 曾静 尚芳红[1] 花雷[1] 阳勇[1,3] 张小梅 WEI Jiang-ping;ZHAO Zi-xuan;ZENG Jing;SHANG Fang-hong;HUA Lei;YANG Yong;ZHANG Xiao-mei(the Third Level Laboratory of Traditional Chinese Medicine Chemistry of the National Administration of Traditional Chinese Medicine,Chongqing Academy of Chinese Materia Medica,Chongqing 400065,China;College of Pharmacy,Chongqing Medical University,Chongqing 400016,China;Luzhou Key Laboratory of Traditional Chinese Medicine for Chronic Diseases Jointly Built by Sichuan and Chongqing,the Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University,Luzhou 646000,China)

机构地区：[1]国家中医药管理局中药化学三级实验室,重庆市中药研究院,重庆400065 [2]重庆医科大学药学院,重庆400016 [3]川渝共建慢病中药新药泸州市重点实验室,西南医科大学附属中医院,四川泸州646000

出　　处：《中国中药杂志》2023年第23期6483-6491,共9页China Journal of Chinese Materia Medica

基　　金：重庆市自然科学基金项目博士后项目(cstc2021jscx-bshX0054);重庆市技术创新与应用发展专项重点项目(cstc2021jscxdxwtBX0012);重庆市中药研究院基本科研业务费项目(jbky20210003)。

摘　　要：探讨异功散预防性给药对D-半乳糖诱导的衰老模型小鼠学习记忆能力的影响及其作用机制,为异功散防治认知下降提供依据。将40只KM小鼠随机分成空白组、模型组、盐酸多奈哌齐组(1.5 mg·kg^(-1))和异功散高剂量组(7.5 g·kg^(-1))、异功散低剂量组(3.75 g·kg^(-1))。除空白组外,其余各组小鼠颈背部注射D-半乳糖(200 g·kg^(-1))复制衰老模型,同时灌胃给予相应药物干预1月后采用水迷宫评价其学习记忆能力改变;苏木精-伊红染色观察海马病理形态学变化;免疫荧光观察海马离子钙结合适配器分子1(ionized calcium binding adapter molecule 1,IBA1)、胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)、趋化因子C-X-C-基元配体12(chemokine C-X-C-motif ligand 12,CXCL12)和趋化因子C-X-C-基元受体4(chemokine CX-C-motif receptor 4,CXCR4)蛋白表达及IBA1、GFAP与CXCR4的位置关系;Western blot检测细胞外调节激酶(extracellular regulated kinase,ERK)及其磷酸化(p-ERK)和肿瘤坏死因子受体1(tumor necrosis factor receptor 1,TNFR1)的蛋白表达;酶联免疫吸附测定法检测脑组织谷氨酸、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)含量及血清和脾脏中TNF-α水平。异功散能够明显缩短衰老模型小鼠隐藏平台实验的逃避潜伏期、增加空间探索实验的跨越平台次数和平台象限累积时间,改善海马区神经细胞排列紊乱、细胞间隙增加和细胞变性或死亡病变,降低受损神经评分,减少IBA1和GFAP阳性表达面积和降低脑组织、血清和脾脏TNF-α水平以及脾脏指数,降低海马区CXCL12和CXCR4平均荧光强度以及降低星形胶质细胞和小胶质细胞中CXCR4的阳性表达,降低p-ERK/ERK、TNFR1表达及谷氨酸含量。该研究表明异功散预防给药改善D-半乳糖诱导的衰老模型小鼠学习记忆能力的下降与其调节脾脏免疫功能和调控脑内CXCL12/CXCR4信号而减少谷氨酸释放有关,但异功散如何通过�This study aims to explore the effect of preventive administration of Yigong Powder on the learning and memory abilities of the mouse model of aging induced by D-galactose and decipher the underlying mechanism,so as to provide a basis for the application of Yigong Powder in the prevention and treatment of cognitive decline.Forty KM mice were randomized into control,model,donepezil(1.5 mg·kg^(-1)),and high-dose(7.5 g·kg^(-1))and low-dose(3.75 g·kg^(-1))Yigong Powder groups.The mice in other groups except the control group were injected with D-galactose(200 g·kg^(-1))at the back of the neck for the modeling of aging.At the same time,the mice were administrated with corresponding drugs by gavage for one month.Morris water maze was used to examine the learning and memory abilities of the mice.Hematoxylin-eosin staining was employed to observe the pathological and morphological changes of the hippocampus.The immunofluorescence assay was employed to detect the expression of ionized calcium-binding adapter molecule 1(IBA1),glial fibrillary acidic protein(GFAP),chemokine C-X-C-motif ligand 12(CXCL12),chemokine C-X-C-motif receptor 4(CXCR4)in the hippocampus and observe the positional relationship between IBA1,GFAP,and CXCR4.Western blot was employed to determine the protein levels of extracellular regulated kinase(ERK),p-ERK,and tumor necrosis factor receptor 1(TNFR1).Enzyme-linked immunosorbent assay was employed to measure the levels of glutamate and tumor necrosis factor(TNF-α)in the brain tissue and the level of TNF-αin the serum and spleen.Yigong Powder significantly shortened the escape latency,increased the times crossing platforms,and prolonged the cumulative time in quadrants of the aging mice.It alleviated the nerve cell disarrangement,increased intercellular space,and cell degeneration or death in the hippocampus and reduced the pathology score of the damaged nerve.Moreover,Yigong Powder reduced the positive area of IBA1 and GFAP,reduced the levels of TNF-αin the brain tissue,serum,and spleen,and decrea

关 键 词：异功散 衰老 认知下降 CXCL12/CXCR4信号通路 谷氨酸 

分 类 号：R285.5[医药卫生—中药学]