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_(1-40)的影响及机制

Effect and Mechanisms of Danxi Tongfeng Fang on Permeability of Blood Brain Barrier and β-Amyloid_(1-40) of Hippocampus in Alzheimer's Disease Mice

目的:探讨丹溪痛风方(DXTFF)对阿尔茨海默病(AD)小鼠血脑屏障(BBB)通透性和海马β淀粉样蛋白1-40(Aβ1-40)的影响及机制。方法:小鼠随机分成正常组,模型组,多奈哌齐组(0.001 g·kg-1),DGP高、中、低剂量组(52,26,13 g·kg-1)。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。DGP连续治疗35 d后取材。检测小鼠脑含水量和伊文氏蓝(EB)含量,海马电镜结构和脏器指数。采用双抗体夹心法测定海马和血清Aβ1-40,海马晚期糖基化终末产物受体(RAGE)和低密度脂蛋白受体相关蛋白1(LRP1)。采用实时荧光定量聚合酶链式反应(Real-time PCR)检测海马RAGE和LRP1 mRNA表达。结果:与正常组比较,模型组AD小鼠脑含水量,EB含量,海马Aβ1-40含量及海马RAGE mRNA水平明显升高(P<0.05),血清Aβ1-40,海马LRP1 mRNA,脾指数和胸腺指数均明显降低(P<0.05),海马CA1区神经元形态较规则,神经元较小,细胞核圆形且萎缩,双层核膜不清晰,细胞质不丰富,细胞器较少;与模型组比较,DGP组脑含水量,EB含量,海马Aβ1-40及RAGE mRNA水平明显降低(P<0.05),血清Aβ1-40,海马LRP1,脾指数和胸腺指数明显增加(P<0.05),海马神经元结构明显改善。结论:DGP通过下调海马RAGE和上调海马LRP1表达来降低AD小鼠BBB通透性和海马Aβ1-40。

Objective： To explore the effect and relevant mechanisms of Danxi Tongfeng Fang（DXTFF） on the permeability of blood brain barrier（BBB） and β-amyloid1-40（Aβ1-40） of hippocampus in Alzheimer＇s disease（AD） mice. Method： Mice were randomly divided into normal group,model group,donepezil（0. 001 g·kg- 1） group,and DXTFF high-dose（52 g·kg- 1）,medial-dose（26 g·kg- 1） and low-dose（13 g·kg- 1） groups. Through injection with Aβ1-42 into bilateral hippocampus and intraperitoneal injection with Dgalactose,the AD mice model was established. The mice were killed after continuous treatment for 35 days. Water content and EB level in brain tissue,electron microscopy of hippocampus and organ coefficients of AD mice were measured. Biochemical methods were used to determine the content of Aβ1-40 in hippocampus and serum,the content of receptor of advanced glycation and products（RAGE） and low-density lipoprotein receptor related protein 1（LRP1） in hippocampus. Real-time PCR methods were used to determine the expression of RAGE and LRP1 mRNA in hippocampus. Result： Compared with the normal group,the model group showed significant increases in water content and EB level in brain tissue,content of Aβ1-40 in hippocampus and content of RAGE in hippocampus（P 〈0. 05）,and significant decreases in Aβ1-40 in serum,LRP1 in hippocampus,spleen index and thymus index（P 〈0. 05）,regular neuronic morphology in hippocampus CA1,smaller neurons,round and atrophic nucleus,nuclear double karyotheca,insufficient cytoplasm and fewer organelle. Compared with the model group,in the DXTFF group, water content in brain, EB level in brain, Aβ1-40 and RAGE level in hippocampus significantly decreased（P〈 0. 05）,and Aβ1-40 level in serum,LRP1 level in hippocampus,organ coefficients of splenic and thymus significantly increased（P 〈0. 05）, with notable improvement in neuronal structure of hippocampus. Conclusion： DXTFF reduces the permeability of BBB and Aβ1-40 in hippocampus by down-regulating RAGE and up-regulating LRP1 in hippocampus.