目的 探究咖啡酸苯乙酯（CAPE）對創(chuàng)傷性顱腦損傷（TBI）大鼠的作用及機制。方法 54只SD大鼠隨機分為假手術(shù)組，模型組，CAPE低、高劑量（5、10 mg·kg^-1）組，CAPE （10 mg·kg^-1）+腺相關(guān)病毒陰性對照（oe-NC，1×10⁹ pfu，200 μL）組，CAPE （10 mg·kg^-1）+過表達(dá)LRRK2腺相關(guān)病毒（oe-LRRK2，1×10⁹ pfu，200 μL）組，每組9只。采用改良的Feeney法制備TBI模型，造模后30 min，CAPE和oe-NC、oe-LRRK2均ip給藥，假手術(shù)組和模型組大鼠ip等量溶劑。所有大鼠每天給藥1次，連續(xù)7 d。改良神經(jīng)功能缺損評分（mNSS）評估大鼠神經(jīng)功能缺損程度；轉(zhuǎn)棒實驗評價大鼠綜合運動能力；檢測各組大鼠腦組織含水量；ELISA法檢測血清神經(jīng)元特異性烯醇化酶（NSE）、白細(xì)胞介素（IL）-6、腫瘤壞死因子（TNF）-α和IL-1β水平；實時熒光定量PCR（qRT-PCR）法檢測LRRK2 mRNA表達(dá)；Western blotting檢測LRRK2、p-JNK和JNK蛋白表達(dá)；TUNEL染色檢測大腦皮層細(xì)胞凋亡；FJB染色檢測神經(jīng)元細(xì)胞死亡。結(jié)果 與假手術(shù)組相比，模型組大鼠mNSS、腦組織含水量、大腦皮層細(xì)胞凋亡率、FJB⁺細(xì)胞數(shù)以及血清中NSE、IL-6、TNF-α和IL-1β含量顯著升高（P<0.05），LRRK2 mRNA和LRRK2、p-JNK/JNK蛋白水平顯著升高（P<0.05），轉(zhuǎn)棒時間顯著降低（P<0.05）；與模型組相比，CAPE低、高劑量組大鼠mNSS、腦組織含水量、大腦皮層細(xì)胞凋亡率、FJB+細(xì)胞數(shù)以及血清中NSE、IL-6、TNF-α和IL-1β含量顯著降低（P<0.05），LRRK2 mRNA和LRRK2、p-JNK/JNK蛋白表達(dá)顯著降低（P<0.05），轉(zhuǎn)棒時間顯著升高（P<0.05）；與CAPE+oe-NC組相比，CAPE+oe-LRRK2組大鼠mNSS、腦組織含水量、細(xì)胞凋亡率、FJB+細(xì)胞數(shù)以及血清中NSE、IL-6、TNF-α和IL-1β含量顯著升高（P<0.05），LRRK2 mRNA和LRRK2、p-JNK/JNK蛋白表達(dá)顯著升高（P<0.05），轉(zhuǎn)棒時間顯著降低（P<0.05）。結(jié)論 CAPE可能通過下調(diào)LRRK2表達(dá)抑制JNK通路活化，在TBI中發(fā)揮保護作用。

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

河南省醫(yī)學(xué)科技攻關(guān)計劃聯(lián)合共建項目（LHGJ20190814）