目的 探究咖啡酸苯乙酯（CAPE）对创伤性颅脑损伤（TBI）大鼠的作用及机制。方法 54只SD大鼠随机分为假手术组，模型组，CAPE低、高剂量（5、10 mg·kg^-1）组，CAPE （10 mg·kg^-1）+腺相关病毒阴性对照（oe-NC，1×10⁹ pfu，200 μL）组，CAPE （10 mg·kg^-1）+过表达LRRK2腺相关病毒（oe-LRRK2，1×10⁹ pfu，200 μL）组，每组9只。采用改良的Feeney法制备TBI模型，造模后30 min，CAPE和oe-NC、oe-LRRK2均ip给药，假手术组和模型组大鼠ip等量溶剂。所有大鼠每天给药1次，连续7 d。改良神经功能缺损评分（mNSS）评估大鼠神经功能缺损程度；转棒实验评价大鼠综合运动能力；检测各组大鼠脑组织含水量；ELISA法检测血清神经元特异性烯醇化酶（NSE）、白细胞介素（IL）-6、肿瘤坏死因子（TNF）-α和IL-1β水平；实时荧光定量PCR（qRT-PCR）法检测LRRK2 mRNA表达；Western blotting检测LRRK2、p-JNK和JNK蛋白表达；TUNEL染色检测大脑皮层细胞凋亡；FJB染色检测神经元细胞死亡。结果 与假手术组相比，模型组大鼠mNSS、脑组织含水量、大脑皮层细胞凋亡率、FJB⁺细胞数以及血清中NSE、IL-6、TNF-α和IL-1β含量显著升高（P<0.05），LRRK2 mRNA和LRRK2、p-JNK/JNK蛋白水平显著升高（P<0.05），转棒时间显著降低（P<0.05）；与模型组相比，CAPE低、高剂量组大鼠mNSS、脑组织含水量、大脑皮层细胞凋亡率、FJB+细胞数以及血清中NSE、IL-6、TNF-α和IL-1β含量显著降低（P<0.05），LRRK2 mRNA和LRRK2、p-JNK/JNK蛋白表达显著降低（P<0.05），转棒时间显著升高（P<0.05）；与CAPE+oe-NC组相比，CAPE+oe-LRRK2组大鼠mNSS、脑组织含水量、细胞凋亡率、FJB+细胞数以及血清中NSE、IL-6、TNF-α和IL-1β含量显著升高（P<0.05），LRRK2 mRNA和LRRK2、p-JNK/JNK蛋白表达显著升高（P<0.05），转棒时间显著降低（P<0.05）。结论 CAPE可能通过下调LRRK2表达抑制JNK通路活化，在TBI中发挥保护作用。

Objective To explore the effects of caffeic acid phenethyl ester (CAPE) on rats with traumatic brain injury (TBI) and its possible mechanism. Methods Totally 54 SD rats were randomly divided into sham operation group, model group, CAPE low and high dose (5, 10 mg·kg^-1) group, CAPE (10 mg·kg^-1) + adeno-associated virus negative control (oe-NC, 1×10⁹ pfu, 200 μL) group, CAPE (10 mg·kg^-1) + over expression of LRRK2 adeno-associated virus (oe-LRRK2, 1×10⁹ pfu, 200 μL) group, nine in each group. TBI model was prepared by modified Feeney method, 30 min after modeling, CAPE, oe-NC, oe-LRRK2 were all administered with ip, and rats of sham operation group and model group was ip the same amount of solvent. All rats were administered once a day for seven days. The neurological function score (mNSS) was used to assess the degree of neurological deficit in rats; the rotating rod test was used to evaluate the comprehensive exercise capacity of rats; The brain water content of rats in each group was measured; ELISA was used to detect the levels of NSE, IL-6, and TNF- α and IL-1β in serum; qRT-PCR was used to detect the expression of LRRK2 mRNA; Western blotting was used to detect the expression of LRRK2, p-JNK and JNK protein; TUNEL staining was used to detect neuronal cell apoptosis; FJB staining was used to detect the neuronal cell death. Results Compared with the sham operation group, the scores of neurological deficit, brain water content, apoptosis rate, number of FJB⁺ cells, and levels of serum NSE, IL-6, TNF-α and IL-1β in the model group were significantly increased (P < 0.05), expressionsn of LRRK2 mRNA, LRRK2 and p-JNK/JNK protein were significantly increased (P < 0.05), and the time of rod transfer was significantly reduced (P < 0.05); compared with model group, the neurological deficit score, brain water content, cell apoptosis rate, number of FJB+ cells, and levels of serum NSEIL-6, TNF- α and IL-1β in CAPE group were significantly reduced (P < 0.05), expressions of LRRK2 mRNA And LRRK2 and p-JNK/JNK protein was significantly reduced (P < 0.05), and the rod time was significantly increased (P < 0.05). Compared with CAPE + oe-NC group, the neurological deficit score, brain water content, apoptosis rate, number of FJB+ cells, and levels of serum NSE, IL-6, TNF- α and IL-1β in CAPE + oe-LRRK2 group increased significantly (P < 0.05), the expressions of LRRK2 mRNA, LRRK2 and p-JNK/JNK protein was significantly increased (P < 0.05), and the rod-rotating time was significantly reduced (P < 0.05). Conclusion CAPE may inhibit the activation of JNK pathway by down-regulating the expression of LRRK2 and play a protective role in TBI.

R285.5

河南省医学科技攻关计划联合共建项目（LHGJ20190814）