摘要
研究开心散对化疗小鼠行为学和脑组织的影响,以探究开心散治疗荷瘤小鼠化疗性认知障碍的作用机制.将30只雌性BALB/c小鼠通过乳腺脂肪垫接种4T1乳腺癌细胞建立荷瘤小鼠模型,并随机均分为肿瘤组、阿霉素组和开心散组.通过旷场实验、高架十字迷宫实验、强迫游泳实验、悬尾实验、Morris水迷宫、新物体识别实验等行为学检测,苏木素-伊红染色、尼氏染色、固蓝染色、Fluoro-Jade B染色、末端脱氧核苷酸转移酶介导的dUTP原位切口末端标记、免疫荧光染色、透射电子显微镜等病理检测,运用网络药理学和全转录组学测序等方法分析化疗性认知障碍产生的机制以及开心散的作用靶点.结果显示,开心散可以防止化疗引起的行为学改变(P＜0.001),增加旷场实验总运动路程和中央区停留时间比,增加高架十字迷宫实验开臂区探索时间,减少强迫游泳实验和悬尾实验静止时间,减少Morris水迷宫逃逸潜伏期和增加穿越平台次数,提高新物体识别实验认知指数;抑制化疗引起的前额叶皮质神经炎症、细胞凋亡、细胞自噬等组织损伤;并重塑化疗过程中前额叶皮质组织的RNA表达谱(P＜0.05).综上研究表明,开心散可能通过重塑化疗过程中前额叶皮质组织的RNA表达谱,减轻神经元组织损伤,最终缓解荷瘤小鼠的化疗性认知障碍.
Abstract
The study aims to evaluate the effect of Kaixin Powder(KXP)on the behavior and brain tissue of chemotherapy-treated mice to explore its mechanism in alleviating chemotherapy-induced cognitive impairment in tumor-bearing mice.Thirty female BALB/c mice were inoculated with 4T1 breast cancer cells to establish a tumor-bearing mouse model and randomly divided into the tumor group,a doxorubicin group,and a KXP group.Behavioral tests,including open field test,elevated plus maze test,forced swimming test,tail suspension test,Morris water maze test,and novel object recognition test,were conducted.Pathological examinations,including hematoxylin-eosin staining,Nissl staining,toluidine blue staining,Fluoro-Jade B staining,terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)assay,immunofluorescence staining,and transmission electron microscopy,were performed.Network pharmacology and whole transcriptome sequencing methods were used to analyze the mechanism of chemotherapy-induced cognitive impairment and the targets of KXP.The results showed that KXP prevented chemotherapy-induced behavioral changes(P＜0.001),increased the total movement distance and central zone residence time in the open field test,increased exploration time in the open arm area in the elevated plus maze test,reduced immobility time in the forced swimming test and tail suspension test,reduced escape latency in the Morris water maze test and increased platform crossings,and improved cognitive index in the novel object recognition test.KXP also inhibited chemotherapy-induced neuroinflammation,apoptosis,and autophagy in the prefrontal cortex,and reshaped the RNA expression profile of the prefrontal cortex tissue during chemotherapy(P＜0.05).In conclusion,KXP may alleviate chemotherapy-induced cognitive impairment in tumor-bearing mice by reshaping the RNA expression profile of prefrontal cortex tissue,thereby reducing neuronal tissue damage.
基金项目
国家自然科学基金项目(81873155)
国家自然科学基金项目(81974571)
国家自然科学基金项目(82274513)
国家自然科学基金项目(82305234)
广东省自然科学基金项目(2020A15110760)
广东省自然科学基金项目(2023A1515011115)
广州市科技计划项目(SL2023A03J01120)
广州市科技计划项目(SL2023A04J00228)
广东省中医院中医药科技专项(YN2022QN32)
广东省中医证候临床研究重点实验室项目(YN2023ZH10)
中医湿证国家重点实验室自主项目(QZ2023ZZ13)
第七批全国老中医药专家学术经验继承工作项目()
NETL
NSTL
万方数据