梓醇-川芎嗪方干预APPswe/PS1dE9双转基因阿尔茨海默病模型小鼠的作用机制研究

Study on Mechanism of Catalpol-Tetramethylpyrazine Prescription on APPswe/PS1dE9 Double Transgenic Mice

目的:探究梓醇-川芎嗪方干预APPswe/PS1dE9双转基因阿尔茨海默病模型小鼠的作用机制。方法:将3月龄C57BL/6J野生型小鼠为对照组(control组),构建阿尔茨海默病模型后,将同月龄APPswe/PS1dE9双转基因小鼠随机分为模型组(model组)、梓醇-川芎嗪方低剂量组(CT-L组)、梓醇-川芎嗪方中剂量组(CT-M组)、梓醇-川芎嗪方高剂量(CT-H组)、安理申组,对照组及模型组均给予生理盐水灌胃,各用药组以相应药物灌胃,每日灌胃1次,连续8周。对各组小鼠进行行为学观察,检测各组小鼠海马组织乙酰胆碱酯酶(AChE)、诱导型一氧化氮合酶(iNOS)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)及血清中β淀粉样蛋白42(Aβ42)。结果:行为学观察:与model组比较,CT-M组、CT-H组、Arceipt组长时程学习和参考记忆能力、自主探究能力、情景识别记忆能力、短期工作记忆能力、条件恐惧记忆能力,缓解焦虑、抑郁状态等7项行为学指标均明显好转(P<0.01);与CT-L组比较,CT-M组、CT-H组、Arceipt组7项行为学指标均明显好转(P<0.05或P<0.01);与CT-M组比较,CT-H组7项行为学指标均明显好转(P<0.05);与Arceipt组比较,CT-H组7项行为学指标均明显好转(P<0.05)。生化检测方面:与model组比较,CT-M组、CT-H组、Arceipt组海马组织SOD、GSH-Px水平均升高,AChE活性、iNOS、IL-6、TNF-α、MDA水平及血清Aβ42水平均降低,差异均有统计学意义(P<0.01);与CT-L组比较,CT-M组、CT-H组、Arceipt组海马组织SOD、GSH-Px水平均升高,AChE活性、iNOS、IL-6、TNF-α、MDA水平及血清Aβ42水平均降低,差异均有统计学意义(P<0.05或P<0.01);与CT-M组比较,CT-H组海马组织SOD、GSH-Px水平均升高(P<0.05),AChE活性、iNOS、IL-6、TNF-α、MDA水平及血清Aβ42水平均降低(P<0.05);与Arceipt组比较,CT-H组海马组织SOD、GSH-Px水平均升高(P<0.05),AChE活性、iNOS、IL-6、TNF-α、MDA水平及血清Aβ42水平均降低(P<0.05)。结论:梓醇-川芎嗪方可以改善阿尔茨海默病模型小鼠的长时程学习和参考记忆能力、自主探究能力、情景识别记忆能力、短期工作记忆能力、条件恐惧记忆能力,缓解焦虑、抑郁状态,且其效应呈剂量依赖性,高剂量梓醇-川芎嗪方优于安理申。其作用机制可能与提高海马组织SOD、GSH-Px水平,降低AChE活性及iNOS、IL-6、TNF-α、MDA、Aβ42水平有关。

Objective:To investigate the effects of Catalpo-l Tetramethylpyrazine Prescription(CT)on APPswe/PS1dE9 double transgenic mice.Methods:Three-month-old C57BL/6J wild-type mice were set as control group,and three-month-old APPswe/PS1dE9 double transgenic mice were divided into model group,Catalpo-l Tetramethylpyrazine Prescription low-dose(CT-L)group,medium-dose(CT-M)group,high-dose(CT-H)group,and Aricept group,respectively.The mice in control group and model group were given intragastric administration of normal saline,and the mice in each medication group were given intragastric administration of the corresponding drug once a day for 8 weeks.The behavior of mice in each group was observed,and the acetylcholinesterase(AChE),inducible nitric oxide synthase(iNOS),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),and malondialdehyde(MDA)in the hippocampal tissues and serum amyloid beta 42(Aβ42)of mice in each group were detected.Results:Behavioral test,Compared with model group,the ability of long-term learning and reference memory,independent inquiry ability,episodic recognition and memory ability,short-term working memory ability,conditional fear memory ability,and anxiety and depression state of mice in CT-M group,CT-H group,and Arceipt group were significantly improved(P<0.01).Compared with CT-L group,the seven behavioral indexes of mice in CT-M group,CT-H group and Arceipt group were improved(P<0.05 or P<0.01).Compared with CT-M group,the seven behavioral indexes of CT-H group were improved(P<0.05).Compared with Arceipt group,the seven behavioral indexes of CT-H group were improved(P<0.05).Biochemical tests,Compared with model group,the levels of SOD and GSH-Px in hippocampus of CT-M group,CT-H group,and Arceipt group were increased,while the activity of AChE,the levels of iNOS,IL-6,TNF-α,and MDA in hippocampus and the level of serum Aβ42 were decreased(P<0.05 or P<0.01).Compared with CT-L group,the levels of SOD and GSH-Px were increased in hippocampus of CT-M group,CT-H group,and Arceipt group,while the activity of AChE,the levels of iNOS,IL-6,TNF-α,and MDA in hippocampus and the level of serum Aβ42 were decreased(P<0.05 or P<0.01).Compared with CT-M group,the levels of SOD and GSH-Px in hippocampus of CT-H group were increased,the activity of AChE,the levels of iNOS,IL-6,TNF-α,and MDA in hippocampus and the level of serum Aβ42 were decreased(P<0.05).Compared with Arceipt group,the levels of SOD and GSH-Px in hippocampus of CT-H group were increased,the activity of AChE,the levels of iNOS,IL-6,TNF-α,and MDA in hippocampus and the level of serum Aβ42 were decreased(P<0.05).Conclusion:Catalpo-l Tetramethylpyrazine Prescription can improve AD model mice long process learning and reference memory ability,the ability of independent exploring,scene recognition memory,short-term memory ability,condition of fear memory ability,reduce anxiety,depression,and its effect is dose-dependent and high-dose Catalpo-l Tetramethylpyrazine Prescription is superior to Aricept.Its mechanism may be related to increasing the levels of SOD and GSH-Px,decreasing the activity of AChE and the levels of iNOS,IL-6,and TNF-αin hippocampus and decreasing the level of MDA in hippocampus and the level of serum Aβ42.