丙戊酸钠对创伤性颅脑损伤大鼠脑组织中金属基质蛋白-9及水通道蛋白4表达的影响  

Valproic acid inhibits the expressions of MMP-9 and AQP-4 proteins in the brain tissue of rats with traumatic brain injury

作　　者：汪棋笙 丁华强 廖帅 何济民 张烨 侯黎明[2] 刘亮 WANG Qi-sheng;DING Hua-qiang;LIAO Shuai;HE Ji-min;ZHANG Ye;HOU Li-ming;LIU Liang(Department of Neurosurgery,Affiliated Hospital of Southwest Medical University,Luzhou 646000,Sichuan,China)

机构地区：[1]西南医科大学附属医院神经外科,泸州646000 [2]合江县人民医院神经外科,泸州646200 [3]合江县人民医院神经外科工作

出　　处：《医学研究生学报》2019年第8期809-814,共6页Journal of Medical Postgraduates

基　　金：泸州市科技基金(2017LZXNYD-J10)

摘　　要：目的大鼠创伤性颅脑损伤模型中,丙戊酸钠(VPA)能否通过降低脑损伤后金属基质蛋白-9(MMP-9)、水通道蛋白4(AQP4)表达发挥脑保护作用目前报道较少。文章研究不同剂量VPA对实验性大鼠创伤性颅脑损伤后的神经保护作用,探讨最佳的给药剂量。方法取成年雄大鼠100只,采用控制性皮层撞击损伤模型(CCI)法建立大鼠创伤性颅脑损伤模型,随机数字表法分成5组假手术组(假手术组与造模组程序一样,但不行硬膜外撞击),外伤模型组[创伤性颅脑损伤(TBI)]、VPA低剂量组(TBI+30mgVPA)、VPA中浓度组(TBI+150mgVPA)、VPA高浓度组(TBI+300mgVPA),每组20只。大鼠于术后24h、3d、7d、14d时间点,分别行外伤功能损伤评分,评分后抽静脉血测血药浓度,抽血后处死,采用干湿重法行脑组织含水测定,Westernblot及免疫组化法测MMP9、AQP4含量。结果TBI组24h、3d、7d、14dmNSS评分分别为(8.4±0.9)、(7.0±0.7)、(5.8±1.0)和(4.5±1.3)分,VPA高浓度组分别为(4.6±1.3)、(3.8±1.3)、(3.0±0.7)和(1.8±0.8)分;与TBI组相比,伤后24h、3d、7d、14d,VPA中、高浓度组的mNSS评分均下降(P<0.05),VPA各浓度组各时间点mNSS评分逐渐降低,两两比较,差异均有统计学意义(P<0.05)。TBI组伤后24h、3d和7d脑含水量分别为(79.6±0.8)%、(82.6±0.8)%、(78.6±0.7)%,VPA高浓度组分别为(76.2±0.7)%、(76.9±1.7)%、(73.9±1.3)%;伤后24h、3d、7d,与TBI组脑组织含水量相比,VPA中、高浓度组降低(P<0.05),VPA各浓度组各时间点逐渐降低,两两比较,差异均有统计学意义(P<0.05)。伤后24h、3d,与TBI组比较,VPA各浓度组AQP4、MMP-9表达下降,给药浓度越高,下降越明显(P<0.05),两两比较发现,VPA各浓度组组间差异有统计学意义(P<0.05)。伤后24h、3d,各组大鼠脑组织AQP4、MMP-9表达免疫组化显示,TBI组见大量阳性表达细胞数,细胞质棕黄色深染,随着给药浓度增加,阳性表达细胞数下降,细胞质棕黄色变浅。结论VPA可抑制�Objective Few studies are reported on the protective effect of valproic acid(VPA)against traumatic brain injury (TBI)by down-regulating the protein expressions of matrix metallo. proteinase-9(MMP-9)and aquaporin-4(AQP-4)in the brain tissue. This study aimed to investigate the neuroprotective effects of different doses of VPA against TBI in experimental rats. Methods We randomly divided 100 adult male rats into five groups of equal number,sham operation,TBI model,and low-(30 mg),medium-(150 mg)and high-dose(300 mg)VPA treatment. At 1,3,7 and 14 days after modeling by controlled cortex impact,we obtained the modified Neurological Severity Scores(mNSS),measured the VPA concentration in the venous blood,and then killed the rats and harvested the brain tissue for determination of the water content using the dry-wet method and the expressions of MMP-9 and AQP-4 by Western blot and immunohistochemistry. Results At 1,3,7 and 14 days after modeling,the mNSSs in the high-dose VPA group were 4.6 ± 1.3,3.8 ± 1.3,3.0 ± 0.7 and 1.8 ± 0.8,respectively,significantly lower than 8.4 ± 0.9,7.0 ± 0.7,5.8 ± 1.0 and 4.5 ± 1.3 in the TBI group(P < 0.05),decreasing in a time-dependent manner, with statistically significant difference between any two dose groups(P < 0.05). At 1,3 and 7 days,the water contents in the brain tissue were(76.2 ± 0.7)%,(76.9 ± 1.7)% and(73.9 ± 1.3)% in the high-dose VPA group,significantly lower than(79.6 ± 0.8)%,(82.6 ± 0.8)% and(78.6 ± 0.7)% in the TBI group(P < 0.05),also decreasing in a time-dependent manner,with statistically significant difference between any two dose groups(P < 0.05). At 1 and 3 days,the expressions of MMP-9 and AQP-4 in the brain tissue were markedly down-regulated in the VPA groups in a dose-dependent manner as compared with those in the TBI group(P < 0.05), with statistically significant difference between any two dose groups(P < 0.05),and meanwhile immunohistochemistry showed large numbers of cells with positive expressions of MMP-9 and AQP-4,which were reduced with th

关 键 词：大鼠颅脑损伤模型 丙戊酸钠 金属基质蛋白9 水通道蛋白4 脑保护作用 

分 类 号：R74[医药卫生—神经病学与精神病学]