SIRT3激动剂厚朴酚通过抑制海马神经元铁死亡缓解小鼠术后认知功能障碍  

Honokiol,an SIRT3 activator,alleviates postoperative cognitive dysfunc⁃tion via inhibiting hippocampal neuronal ferroptosis in mice

作　　者：黄涛 韩甜甜 许倩倩 张振 胡鹏超 丁旭东 罗辉宇 曾炼 HUANG Tao;HU Pengchao;HAN Tiantian;XU Qianqian;ZHANG Zhen;DING Xudong;LUO Huiyu;ZENG Lian(Xiangyang No.1 People's Hospital,Hubei University of Medicine,Wuhan 430000,China;Department of Anesthesiolo-gy,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430000,China)

机构地区：[1]湖北医药学院附属襄阳市第一人民医院,湖北武汉430000 [2]华中科技大学同济医学院附属同济医院麻醉科,湖北武汉430000

出　　处：《中国病理生理杂志》2024年第5期827-835,共9页Chinese Journal of Pathophysiology

基　　金：湖北省教育厅科研计划重点项目(No.D20212103);湖北省卫健委项目(No.WJ2023F077;No.WJ2023F078);襄阳市科技局项目(No.2022YL29A;No.2022YL24B;No.2021YL123;No.2021ZD12);湖北医药学院研究生创新项目(No.YC2022048);襄阳市第一人民医院科技创新项目(No.XYY2023SD05;No.XYY2023SD21;No.XYY2023QT12)。

摘　　要：目的:探究沉默信息调节因子3(SIRT3)激动剂厚朴酚(HKL)在小鼠术后认知功能障碍(POCD)中的作用及其可能机制。方法:将10月龄雄性C57/BL6小鼠随机分成对照(Con)组、手术(Sur)组和Sur+HKL组(n=10)。Sur+HKL组小鼠给予HKL腹腔注射预处理7 d,Sur和Sur+HKL组小鼠均采用胫骨骨折切开复位内固定术的方法建立POCD模型。使用旷场实验、新物体识别实验、Morris水迷宫实验和Y迷宫实验来检测小鼠的认知功能;尼氏染色评价小鼠海马和皮层神经元形态、结构和活力;Western blot检测小鼠海马谷胱甘肽过氧化物酶4(GPX4)、溶质载体家族7成员11(SLA7A11)、酰基辅酶A合成酶长链家族成员4(ACSL4)、SIRT3和核因子E2相关因子2(NRF2)蛋白的表达;免疫荧光染色测定GPX4表达水平。结果:与Con组比较,Sur组和Sur+HKL组小鼠在旷场实验中移动的总距离和中央区域探索时间差异无统计学意义(P>0.05);新物体识别实验和Y迷宫实验结果显示,Sur组小鼠识别指数和轮替次数较Con组显著降低(P<0.01),侧臂封锁再通后,小鼠进入新臂的距离百分比和次数百分比显著下降(P<0.01);水迷宫训练时Sur组小鼠潜伏期较Con组下降缓慢,测试时Sur组小鼠潜伏期、目的象限穿越次数及穿越时间百分比均显著低于Con组(P<0.01);与Sur组比较,Sur+HKL组上述指标均显著升高(P<0.01)。尼氏染色结果显示,Sur组小鼠海马CA1区和内侧前额叶皮层的神经元染色较浅,尼氏染色阳性神经元数量显著下降(P<0.01);HKL治疗后神经元活力明显增强。Western blot结果显示,与Con组比较,Sur组SIRT3、GPX4、SLC7A11和NRF2蛋白表达水平显著降低(P<0.05),ACLS4蛋白表达水平显著升高(P<0.05);与Sur组比较,Sur+HKL组上述结果被逆转,差异有统计学意义(P<0.05)。海马神经元的免疫荧光染色与Western blot结果一致,Sur组海马区神经元中GPX4的表达水平显著下降,HKL显著升高神经元GPX4的表达水平(P<0.01)。结论:SIRT3AIM:To investigate the impact of honokiol(HKL),an activator of silent information regulator 3(SIRT3),on postoperative cognitive dysfunction(POCD)in mice,and to explore the potential mechanisms.METHODS:Ten-month-old male C57/BL6 mice were randomly divided into control(Con)group,surgical(Sur)group and Sur+HKL group(n=10).The mice in Sur+HKL group were intraperitoneally injected with HKL for 7 d before modeling.The mice in Sur and Sur+HKL groups underwent tibial fracture open reduction and internal fixation to establish the POCD model.The assessment of cognitive function was conducted using the open-field test(OFT),novel object recognition test(NORT),Morris water maze test(MWMT),and Y-maze test(YMT).Nissl staining was employed to assess the morphology,struc-ture and vitality of hippocampal and cortical neurons in mice.The protein expression of glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11),acyl coenzyme A synthetase long-chain family member 4(ACSL4),SIRT3 and nuclear factor E2-related factor 2(NRF2)in the mouse hippocampus was detected by Western blot,while im-munofluorescence staining was utilized to determine GPX4 level in mouse neurons.RESULTS:No statistically signifi-cant differences were observed among the groups in terms of total distance moved and central zone exploration during the OFT(P>0.05).However,the results from the NORT and YMT indicated that the mice in Sur group exhibited significant-ly lower recognition indexes,reduced alternation rates(P<0.01),and decreased percentages of entries and crossing time into the new arm after side arm blockade(P<0.01),when compared with Con group.Furthermore,the mice in Sur group demonstrated a slower decrease in latency during the learning period of MWMT,while significantly lower latency,fewer crossing number and lower percentage of time in the target quadrant were observed during the testing period of MWMT(P<0.01).The above indicators were obviously enhanced in Sur+HKL group compared with Sur group(P<0.01).The results of Nissl staining indic

关 键 词：厚朴酚 术后认知功能障碍 铁死亡 沉默信息调节因子3 

分 类 号：R742[医药卫生—神经病学与精神病学] R619[医药卫生—临床医学] R363.2