芒柄花黄素对阿尔茨海默病小鼠血脑屏障通透性和海马闭锁小带蛋白-1表达的影响
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摘要
目的探讨芒柄花黄素(FMN)对阿尔茨海默病(AD)小鼠血脑屏障(BBB)通透性和海马闭锁小带蛋白(ZO)-1表达的影响。方法将雄性小鼠随机分成对照组、模型组、多奈哌齐组(1 mg/kg)、FMN高、中、低剂量组(30.0、15.0、7.5 mg/kg)。采用小鼠双侧海马注射Aβ1～42和腹腔注射D-半乳糖诱导AD小鼠。FMN连续治疗35 d后取材,检测小鼠脑含水量和伊文思蓝(EB)含量,采用免疫组化、实时定量聚合酶链反应(PCR)和蛋白质印迹检测海马ZO-1表达。结果与对照组比较,模型组BBB通透性明显增加(P<0.05),海马ZO-1表达明显降低(P<0.05)。与模型组比较,FMN高、中剂量组BBB通透性明显降低(P<0.05),海马ZO-1表达明显增加(P<0.05)。结论 FMN通过上调海马ZO-1表达来降低AD小鼠BBB通透性。
Objective To explore the effects of formononetin(FMN) on blood brain barrier(BBB) and zonula occludens(ZO)-1 of the hippocampus in Alzheimer′s disease(AD) mice.Methods Male mice were randomly divided into control,model,donepezil(1 mg/kg),FMN high-dose(30.0 mg/kg),FMN medial-dose(15.0 mg/kg),FMN low-dose(7.5 mg/kg) groups. Microinjection incubated Aβ1～42 into the dorsal blade of the dentale gyrus in the bilateral hippocampus combined with intraperitoneal injection of D-galactose was used to induce AD model. The mice were killed after 35 days continuous treatment. After the treatment, water content and EB level in brain tissue were detected. Immunohistochemistry, real time-PCR and Western blot were used to determine the content of ZO-1 in hippocampus.Results Compared with the control group,the permeability of BBB in the model group was significantly increased(P<0.05), the expression of ZO-1 in the hippocampus was significantly decreased(P<0.05). Compared with the model group, FMN high-dose and medial-dose could significantly improve the permeability of BBB of AD mice(P<0.05),could significantly increase ZO-1 in the hippocampus(P<0.05).Conclusions FMN should depress the damage of BBB in hippocampus of AD mice. FMN plays a certain role in the treatment of AD through inhibiting ZO-1.
Objective To explore the effects of formononetin(FMN) on blood brain barrier(BBB) and zonula occludens(ZO)-1 of the hippocampus in Alzheimer′s disease(AD) mice.Methods Male mice were randomly divided into control,model,donepezil(1 mg/kg),FMN high-dose(30.0 mg/kg),FMN medial-dose(15.0 mg/kg),FMN low-dose(7.5 mg/kg) groups. Microinjection incubated Aβ1～42 into the dorsal blade of the dentale gyrus in the bilateral hippocampus combined with intraperitoneal injection of D-galactose was used to induce AD model. The mice were killed after 35 days continuous treatment. After the treatment, water content and EB level in brain tissue were detected. Immunohistochemistry, real time-PCR and Western blot were used to determine the content of ZO-1 in hippocampus.Results Compared with the control group,the permeability of BBB in the model group was significantly increased(P<0.05), the expression of ZO-1 in the hippocampus was significantly decreased(P<0.05). Compared with the model group, FMN high-dose and medial-dose could significantly improve the permeability of BBB of AD mice(P<0.05),could significantly increase ZO-1 in the hippocampus(P<0.05).Conclusions FMN should depress the damage of BBB in hippocampus of AD mice. FMN plays a certain role in the treatment of AD through inhibiting ZO-1.
引文
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7 Liao F,Meng Y,Zheng H,et al.Biospecific isolation and characterization of angiogenesis-promoting ingredients in Buyang Huanwu decoction using affinity chromatography on rat brain microvascular endothelial cellscombined with solid-phase extraction and HPLC-MS/MS[J].Talanta,2018;(179):490-500.
8 张英博,费洪新,郭家,等.蝙蝠葛碱对阿尔茨海默病小鼠海马晚期糖基化终产物受体和核转录因子-κBp65的影响[J].中国老年学杂志,2017;37(19):4697-700.
9 费洪新,高音,张英博,等.蝙蝠葛苏林碱对阿尔茨海默病小鼠海马的影响和机制[J].中国老年学杂志,2017;37(1):9-12.
10 张晓杰,费洪新,刘得水,等.丹溪痛风方对阿尔茨海默病小鼠血脑屏障通透性和海马β淀粉样蛋白的影响及机制[J].中国实验方剂学杂志,2016;22(22):118-23.
2 Yin T,Xie W,Sun J,et al.Penetratin peptide-functionalized gold nanostars:enhanced BBB permeability and NIR photothermal treatment of Alzheimer′s disease using ultralow irradiance[J].ACS Appl Mater Interfaces,2016;8(30):19291-302.
3 Bien-Ly N,Boswell CA,Jeet S,et al.Lack of Widespread BBB disruption in Alzheimer′s disease models:focus on therapeutic antibodies[J].Neuron,2015;88(2):289-97.
4 王利,丁晓瑜,毕明俊,等.正丁基甲硫醚对急性一氧化碳中毒大鼠血脑屏障ZO-1和claudin-5表达的影响[J].中华危重病急救医学,2018;30(5):422-7.
5 Zhang S,An Q,Wang T,et al.Autophagy and MMP-2/9-mediated reduction and redistribution of ZO-1 contribute to hyperglycemia-increased blood-brain barrier permeability during early reperfusion in stroke[J].Neuroscience,2018;(377):126-37.
6 Fei HX,Zhang YB,Liu T,et al.Neuroprotective effect of formononetin in ameliorating learning and memory impairment in mouse model of Alzheimer′s disease[J].Biosci Biotechnol Biochem,2018;82(1):57-64.
7 Liao F,Meng Y,Zheng H,et al.Biospecific isolation and characterization of angiogenesis-promoting ingredients in Buyang Huanwu decoction using affinity chromatography on rat brain microvascular endothelial cellscombined with solid-phase extraction and HPLC-MS/MS[J].Talanta,2018;(179):490-500.
8 张英博,费洪新,郭家,等.蝙蝠葛碱对阿尔茨海默病小鼠海马晚期糖基化终产物受体和核转录因子-κBp65的影响[J].中国老年学杂志,2017;37(19):4697-700.
9 费洪新,高音,张英博,等.蝙蝠葛苏林碱对阿尔茨海默病小鼠海马的影响和机制[J].中国老年学杂志,2017;37(1):9-12.
10 张晓杰,费洪新,刘得水,等.丹溪痛风方对阿尔茨海默病小鼠血脑屏障通透性和海马β淀粉样蛋白的影响及机制[J].中国实验方剂学杂志,2016;22(22):118-23.