基于SIRT1/NRF2/HO-1信号探讨樱草杜鹃石油醚提取物抗痴呆的作用及机制  

Study on the Anti-dementia Effect and Its Mechanism of Petroleum Ether Extract from Rhododendron Primulaeflorum Bur. Et Franch. Based on SIRT1/NRF2/HO-1 Signaling Pathway

作　　者：屈培珍 魏江平 刘毅[3] 瞿显友[1] 王云红[1,2] 刘翔 高思佳[1] 樵星芳 阳勇[1,2] 张小梅[1,2] 钟国跃 QU Peizhen;WEI Jiangping;LIU Yi;ZHAI Xianyou;WANG Yunhong;LIU Xiang;GAO Sijia;QIAO Xingfang;YANG Yong;ZHANG Xiaomei;ZHONG Guoyue(Chongqing Academy of Chinese Materia Medica,Chongqing 400065,China;School of Pharmacy,Zunyi Medical University,Zunyi 563003 Guizhou,China;Chongqing University of Posts and Telecommunications,Chongqing 400065,China;Research Center of Chinese Medicine Resources and Ethnic Minority Medicine,Jiangxi University of Chinese Medicine,Nanchang 330004 Jiangxi,China)

机构地区：[1]重庆市中药研究院、国家中医药管理局中药化学三级实验室,重庆400065 [2]遵义医科大学药学院,贵州遵义563003 [3]重庆邮电大学,重庆400065 [4]江西中医药大学中药资源与民族药研究中心,江西南昌330004

出　　处：《中药新药与临床药理》2023年第3期341-348,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家重点研发计划“中医药现代化研究”重点专项(2019YFC1712300)。

摘　　要：目的探讨樱草杜鹃石油醚提取物(RPE)对痴呆模型小鼠的学习记忆功能的影响及其潜在的作用机制。方法根据体质量将小鼠随机分成空白组、模型组、安理申组(1.5 mg·kg^(-1))、RPE高(4.5 g·kg^(-1))和低(2.25 g·kg^(-1))剂量组,采用D-半乳糖(100 mg·kg^(-1))颈背部皮下注射制备痴呆模型,造模当天开始灌胃给药(10 mL·kg^(-1)),连续给药30 d。采用Morris水迷宫评价小鼠学习记忆能力变化;苏木精-伊红染色观察小鼠海马病理形态学改变;免疫组化检测沉默信息调节因子2同源体1(SIRT1)的蛋白表达;Western Blot法分析核因子红细胞系2相关因子2(NRF2)、血红素加氧酶1(HO-1)、B细胞淋巴瘤/白血病-2基因(Bcl-2)、BCL2-相关X蛋白(Bax)和半胱氨酸蛋白酶3(Caspase-3)的蛋白表达;生化试剂盒检测血清和脑组织中超氧化物歧化酶(SOD)、谷胱甘肽(GSH)、过氧化氢酶(CAT)活性及过氧化氢(H_(2)O_(2))和丙二醛(MDA)含量。结果与空白组比较,模型组小鼠第5天的逃避潜伏期明显延长(P<0.01),空间探索实验进入平台次数明显减少(P<0.05);海马区神经细胞数量减少、排列紊乱,海马区和皮层均可见较多神经细胞变性或死亡且病理评分明显升高(P<0.01,P<0.001);血清和脑组织SOD、GSH和CAT活性均明显降低(P<0.05,P<0.001)而H_(2)O_(2)(血清)和MDA含量则均明显升高(P<0.05,P<0.001);SIRT1、NRF2、Bcl-2的蛋白表达均下调而Bax和Caspase-3的蛋白表达均上调(P<0.05,P<0.01)。与模型组比较,RPE可明显缩短模型小鼠第5天的逃避潜伏期(P<0.05),明显增加平台象限时间和进入平台次数(P<0.05,P<0.01,P<0.001);改善海马和皮层神经细胞病变及降低病理评分(P<0.05,P<0.01);明显提高模型小鼠血清和海马组织SOD、GSH和CAT活性(P<0.05,P<0.01,P<0.001)而降低H_(2)O_(2)(血清)和MDA含量(P<0.01,P<0.001);上调脑组织SIRT1、NRF2、HO-1、Bcl-2的蛋白表达(P<0.05,P<0.01,P<0.001)而下调Bax和Caspase-3的蛋白表达(P<0.05)。�Objective To investigate the effect of petroleum ether extract from Rhododendron primulaeflorum Bur. et Franch.(RPE) on learning and memory function in dementia mice and its potential mechanism.Methods Mice were randomly divided into blank group,model group,Aricept group(1.5 mg·kg^(-1)),high-(4.5 g·kg^(-1))and lowdose RPE(2.25 g·kg^(-1))groups according to body mass index. Dementia models were prepared by subcutaneous injection of D-galactose(100 mg · kg^(-1)) into the cervical dorsal region. All mice received corresponding drugs(10 mL·kg^(-1))by gavage on the first day of modeling,administered for 30 consecutive days. Morris water maze was used to evaluate the learning and memory ability of mice,the pathomorphological changes in hippocampus of mice were observed by hematoxylin eosin staining. The protein expression of silence information regulator 1(SIRT1)was detected by immunohistochemistry,while the protein expressions of nuclear factor erythrocyte line 2-related factor 2(NRF2),heme oxygenase 1(HO-1),B-cell lymphoma/leukemia 2(Bcl-2),BCL2-associated X protein(Bax)and cysteine proteinase-3(Caspase-3) were analyzed by Western Blot. The activity of superoxide dismutase(SOD),glutathione(GSH),catalase(CAT)and the content of hydrogen peroxide(H_(2)O_(2))and malondialdehyde(MDA)in serum and brain were detected by biochemical kit.Results Compared with the blank group,the escape latency of mice in the model group was significantly prolonged on the fifth day(P<0.01) and the number of entering the platform in the space exploration experiment was significantly reduced(P<0.05). The number of neurons in the hippocampus decreased and the arrangement was disordered. Nerve cell degeneration and death in the cortex and hippocampus were found and their pathological score were significantly increased(P<0.01,P<0.001).The activities of SOD,GSH and CAT in serum and brain were significantly decreased(P 0.05,P<0.001),while the content of H_(2)O_(2)(only in serum)and MDA were significantly increased(P<0.05,P<0.001). The protein ex

关 键 词：樱草杜鹃 石油醚提取物 痴呆症 氧化应激 细胞凋亡 小鼠 

分 类 号：R285.5[医药卫生—中药学]