基于核受体辅激活因子4介导的铁自噬探讨补肾活血颗粒对帕金森病模型小鼠脑黑质神经元铁死亡的影响  

Effects of Bushen Huoxue Granules on ferroptosis in brain nigrostriatal neurons of Parkinson’s disease model mice based on NCOA4-mediated ferritinophagy

作　　者：谌盈帆 郝斐然 汤响林 齐小荣 李绍旦[1] 杨明会[1] CHEN Yingfan;HAO Feiran;TANG Xianglin;QI Xiaorong;LI Shaodan;YANG Minghui(Department of Traditional Chinese Medicine,Sixth Medical Center of Chinese PLA General Hospital,Beijing 100048,China;Medical School of Chinese PLA,Beijing 100853,China;Academy of MilitaryMedical Sciences,Beijing 100850,China)

机构地区：[1]解放军总医院第六医学中心中医学部,北京100048 [2]解放军医学院,北京100853 [3]军事医学研究院,北京100850

出　　处：《中华中医药杂志》2024年第2期926-931,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金重点项目(No.82130117)。

摘　　要：目的:探讨补肾活血颗粒(BSHXG)对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)诱导的帕金森病(PD)模型小鼠铁自噬-铁死亡的影响。方法:将72只小鼠随机分为正常组、模型组、美多芭组和BSHXG低、中、高剂量组,每组12只,采用MPTP腹腔注射的方法诱导PD小鼠模型。各组小鼠采用爬杆和转棒疲劳试验进行行为学评价;透射电镜检测脑黑质细胞线粒体形态;免疫组化检测脑黑质酪氨酸羟化酶(TH)阳性细胞表达;免疫印迹法检测脑黑质核受体辅激活因子4(NCOA4)、铁蛋白重链1(FTH1)、长链脂酰辅酶A合成酶4(ACSL4)、溶血卵磷脂酰基转移酶3(LPCAT3)蛋白的表达;比色法检测脑黑质Fe^(2+)、总超氧化物歧化酶(T-SOD)、谷胱甘肽(GSH)含量。结果:与模型组比较,美多芭、BSHXG各剂量组小鼠爬杆转向时间及总时间显著缩短(P<0.01),转棒停留时间显著增加(P<0.01),脑黑质神经元细胞TH阳性表达增加,线粒体结构形态明显改善,BSHXG各剂量组NCOA4、Fe^(2+)、ACSL4、LPCAT3表达显著降低(P<0.05,P<0.01),BSHXG中、高剂量组FTH1蛋白表达显著升高(P<0.05,P<0.01),美多芭、BSHXG各剂量组T-SOD、GSH表达显著增加(P<0.01,P<0.05)。结论:BSHXG可通过调节NCOA4介导的铁自噬,抑制脑黑质神经元细胞铁死亡,从而改善多巴胺能神经元损伤。Objective:To investigate the effect of Bushen Huoxue Granules(BSHXG)on ferritinophagy-ferroptosis in MPTP-induced Parkinson’s disease(PD)model mice.Methods:Seventy-two mice were randomly divided into normal group,model group,Medopar group and BSHXG high-,medium-and low-dose group,12 mice in each group,and the PD mouse model was induced by intraperitoneal injection of MPTP.The mice in each group were evaluated behaviorally using pole-climbing test and baton twirling fatigue test;transmission electron microscopy was used to detect the mitochondrial morphology of brain nigrostriatal cells;immunohistochemistry was used to detect the expression of brain nigrostriatal tyrosine hydroxylase(TH)-positive cells;immunoblotting was used to detect the expression of brain nigrostriatal NCOA4,FTH1,ACSL4,and LPCAT3 proteins;and colorimetric assay was used to detect the content of brain nigrostriatal Fe^(2+),T-SOD,and GSH.Results:Compared with the model group,the rod-climbing steering time and total time of mice in the Medopar and BSHXG three dose groups were shortened(P<0.01),and the stick-turning residence time was increased(P<0.01),the positive expression of TH in brain nigrostriatal neuronal cells was increased, the structural morphology of the mitochondria was improved, and the expression of NCOA4, Fe^(2+), ACSL4, and LPCAT3 in BSHXG three dose groups were decreased (P<0.05, P<0.01), and the expression of FTH1 in BSHXG medium- and high-dose groups was increased (P<0.05, P<0.01), the expression of T-SOD and GSH in the Medopar and BSHXG three dose groups were increased (P<0.01, P<0.05). Conclusion: BSHXG ameliorates DA ergic neuronal injury by regulating NCOA4-mediated ferritinophagy and inhibiting ferroptosis in brain nigrostriatal neuronal cells.

关 键 词：补肾活血颗粒 帕金森病 铁死亡 铁自噬 核受体辅激活因子4 1-甲基-4-苯基-1 2 3 6-四氢吡啶 机制 

分 类 号：R285.5[医药卫生—中药学]