还脑益聪方对老龄认知障碍大鼠氧化应激和抗衰老指标作用的研究

Effects of HuannaoYicong Decoction on the oxidative stress and anti-aging in aging rats with cognitive impairment

目的观察还脑益聪方对动物行为学、病理形态、氧化应激和衰老指标的影响。方法根据Morris水迷宫实验结果结合体质量,从60只老年大鼠中选出正常老年大鼠,并按照分层随机方法分为老年认知障碍(老年模型)组,阳性对照药(盐酸多奈哌齐片,0.49 mg/kg)组,中药小剂量(3.78 g生药/kg)、中剂量(7.56 g生药/kg)和大剂量(18.90 g生药/kg)组,同时另设10只4月龄SD青年大鼠为青年正常组。老年正常组、老年模型组和青年正常组予等体积蒸馏水灌胃,其他组给予相应的药物灌胃,连续灌胃14周。给药结束后,以Morris水迷宫实验测试大鼠学习记忆能力,腹腔麻醉,腹主动脉取血;按照随机原则选取5只大鼠,并取其脑组织(海马)匀浆,检测血清和组织中超氧化歧化酶(SOD)、丙二醛(MDA)、过氧化氢酶(CAT)、总抗氧化能力(T-AOC)、谷胱甘肽(GSH)等氧化应激相关指标;另外5只大鼠脑组织,HE染色和刚果红染色,观察海马组织CA1区形态学变化和淀粉样沉积变化。结果与老年模型组相比,中药大剂量组可显著提高大鼠穿越第四象限的时间和路程(P﹤0.05,P﹤0.01)。与老年模型组大鼠海马区相比,中药大、中和小剂量干预后其病变程度明显减轻,刚果红染色后镜下可见给药后β-淀粉样蛋白(Aβ)沉积数量明显减少。与老年模型组比较,中药小、中、大剂量及盐酸多奈哌齐均可提高血清和海马组织中SOD活性,降低MDA含量(P﹤0.05,P﹤0.01);中药小、大剂量还可显著提高血清和海马中CAT和GSH活性(P﹤0.01),降低海马中β-AP含量(P﹤0.05,P﹤0.01)。中药中、大剂量组血清中GSH-PX活性有增高趋势,但无统计学意义(P>0.05)。结论自然衰老大鼠学习记忆和空间识别能力减弱,还脑益聪颗粒剂可改善自然衰老大鼠AD模型的学习记忆功能,可保护脑组织海马结构,提高血清及组织中抗氧化能力和T-AOC的活性,减少β-AP的生成。

Objective To observe effects of HuannaoYicong Decoction （HY） on ethology, pathological morphology,oxidative stress and anti-aging in order to initially explore its effect mechanism and action process. Methods Sixty 18 to 20-month-oldSprague-Dawley aging rats, according to results of Morris water maze and body mass, were selected. By stratified randomized method, rats were divided into anaging modelgroup, a positive controlmodicine group（donepezil hydrochloride, 0.49 mg/kg）, a small-dose of HY granulegroup （3.78 g/Kg）, moderate-dose of HYgranulegroup （7.56 g/Kg）, and a large-dose of HYgranulegroup （18.90 g/Kg）. In addition, 10 4-month-old youth rats were taken as youth normal group. Intragastric administration of distilled water was applied in the aging normal group, aging model group and youth group for coniinuous 14 weeks. After the administration, Morris water maze test was performed to measure rats＇ learning-memory ability. With intraperitoneal anesthesia, blood was collected from abdominal aorta; brain tissue （hippocampus）homogenate of 5 rats were randomly selected to measure oxidativestress index, such asSOD, MDA, CAT, T-AOC, GSH.Other 5 rats＇ brain tissues were selected and fixed by 10 % neutral formaldehyde solutionto observe morphological and amyloid deposition changes of hippocampal tissue CA1 region with HE andCongo red staining.Results Compared with aging model group, the large-dose of HY granule group could significantly increase rats＇running time and distancein the fourth quadrant （P 〈 O.05,P 〈 0.01）. Compared with hippocampus area in rats with cognitive impairment, lesion severity was obviously relieved after intervention of HY decoction with small, moderate and large dose. Mieroscopicallythe number of β-amyloid protein deposition was obviouslydecreasedwith Congo red staining.Compared with model group, SOD activity was improved and MDA content was decreased in serum and hippocampus tissue in the positive control medicine group, small-dose group, moderate-dose group and large-dose group.The small-dose group and large-dose group could significantly improveSOD and GSH activity in serum and hippocampus tissue and lower β-AP in hippocampus tissue.There was a rising trend of GSH-PX activity inserum and hippocampus tissuein middle-dose group and large-dose group, which had no statistical significance.Conclusion The learning-memory ability and space recognition capability is impaired in aging rats. HY eould improve aging alzheimer diseaserats＇learning-memory ability, protect hippocampus formation in brain tissue, increase antioxidant capacity and T-AOC activity in serum and tissue, and decrease β-amyloid protein content.