摘要：

目的 探讨姜黄素（curcumin）对颅脑创伤小鼠大脑皮质神经元凋亡、自噬及神经修复的治疗作用。方法 90只健康雄性小鼠随机分为空白对照组（12只）、假模型组（26只）、颅脑创伤+生理盐水组（TBI+NaCl组，26只，模型制备后30min腹腔注射生理盐水）和颅脑创伤+姜黄素组（TBI+ curcumin组，26只，模型制备后30min腹腔注射姜黄素），通过碘化丙啶染色检测神经元凋亡数目、 Westernblotting法检测凋亡和自噬相关蛋白［Ｂ细胞淋巴瘤/白血病-2（Bcl-2）、激活型Caspase-3和P62、 微管相关蛋白1轻链3（MAP1LC3）］表达变化，Cavalieri法测量大脑皮质损伤体积。结果 颅脑创伤后 48h，小鼠神经元凋亡数目显著增加（t=26.000， P=0.000； t=18.520， P=0.000），姜黄素治疗后神经元凋亡数目减少（t=3.686， P=0.014）；大脑皮质损伤区凋亡相关蛋白Bcl-2表达降低（t=5.656， P=0.000； t= 3.254， P=0.008； t=4.485， P=0.001； t=3.150， P=0.009）、激活型Caspase-3表达升高（t=8.534， P=0.000； t=3.468， P=0.005； t=8.565， P=0.000； t=2.846， P=0.016），自噬相关蛋白P62表达降低（t=3.574， P= 0.004； t=7.612， P=0.000； t=3.465， P=0.005； t=8.637， P=0.000）、MAP1LC3-Ⅱ表达升高（t=7.101， P= 0.000； t=7.656， P=0.000； t=6.816， P=0.000； t=9.043， P=0.000），姜黄素治疗后Bcl-2表达升高（t= 3.290， P=0.007）、激活型Caspase-3表达降低（t=3.520， P=0.005）、P62表达持续降低（t=3.595， P= 0.004）、MAP1LC3-Ⅱ表达持续升高（t=4.954， P=0.000）。颅脑创伤后28d，小鼠大脑皮质损伤体积增加 （t=34.813， P=0.000； t=11.172， P=0.000），姜黄素治疗后大脑皮质损伤体积减少（t=4.525， P=0.003）。 结论 姜黄素可通过抑制神经元凋亡活性、增强自噬活性而对颅脑创伤后的神经元发挥保护作用并促进神经修复。

关键词: 颅脑创伤, 姜黄素, 细胞凋亡, 疾病模型, 动物

Abstract:

Objective To explore the therapeutic effect of curcumin on neronal apoptosis, autophagy and neural renovation in cerebral cortex of mice after traumatic brain injury (TBI). Methods A total of 90 mice were randomly divided into 4 groups: blank control group (N=12), sham group (N=26), TBI+NaCl group (N=26, NaCl was pretreated by intraperitoneal injection 30min after TBI) and TBI+ curcumin group (N=26, curcumin was pretreated by intraperitoneal injection 30 min after TBI). Propidium iodide (PI)-labeled cell-counting was used to study the cell protective effect of curcumin. Expression of apoptosis and autophagy related proteins B celllymphoma (Bcl-2), activated Caspase-3, P62, microtubule-associated protein 1 light chain 3 (MAP1LC3-Ⅱ) were detected by Western Blotting. The lesion volume of cortex was tested by Cavalieri mothod. Results PI positive cells significantly increased 48h after TBI (t= 26.000, P=0.000; t=18.520, P=0.000), and reduced after curcumin injection (t=3.686, P=0.014); in cortex lesion area,expression level of apoptosis associated protein Bcl-2 decreased(t=5.656, P=0.000; t= 3.254, P =0.008; t =4.485, P =0.001; t =3.150, P =0.009), expression level of activated Caspase-3 increased (t=8.534, P=0.000; t=3.468, P=0.005; t=8.565, P=0.000; t=2.846, P=0.016), expression level of autophagy associated protein P62 decreased (t=3.574, P=0.004; t=7.612, P=0.000; t=3.465, P= 0.005; t=8.637, P=0.000), expression level of MAP1LC3-Ⅱ increased (t=7.101, P=0.000; t=7.656, P= 0.000; t=6.816, P=0.000; t=9.043, P=0.000), curcumin suppressed apoptotic activity by showing higher expression level of Bcl-2 (t=3.290, P=0.007), lower expression level of activited Caspase-3 (t=3.520, P= 0.005), lower expression level of P62 (t=3.595, P=0.004), and higher expression level of MAP1LC3-Ⅱ (t= 4.954, P=0.000). More tissue damage was seen 28d after TBI (t=34.813, P=0.000; t=11.172, P=0.000) but less tissue damage was seen in TBI+curcumin group (t=4.525, P=0.003). Conclusions Curcumin could inhibit neuronal apoptosis and enhance antophagic activity. Therefore,it may induce neuroprotection and promote nerual renovaion after fraumatic brain injury.

Key words: Craniocerebral trauma, Curcumin, Apoptosis, Disease models, animal