胡黄连苷Ⅱ对大鼠脑缺血性损害和基质金属蛋白酶-9表达的影响及治疗的最佳时间窗与剂量的选取  

Effect of picroside Ⅱ on cerebral ischemic injury and expression of matrix metalloproteinase-9 in rats and the selection of the best time window and dose of treatment

作　　者：张美增[1] 逄芳芳[1] 赵丽[1] 常翠翠[1] 郭云良[1] 

机构地区：[1]青岛大学附属医院神经内科,266003

出　　处：《临床神经病学杂志》2015年第2期114-120,共7页Journal of Clinical Neurology

摘　　要：目的：探讨胡黄连苷Ⅱ对大鼠脑缺血性损伤和基质金属蛋白酶－9（ MMP－9）表达的影响，以及最佳治疗时间窗与剂量的选取。方法将174只Wistar大鼠随机分为脑缺血组、正常对照组和治疗组，按正交试验设计治疗组大鼠再分为缺血1．0 h、1．5 h、2．0 h、2．5 h，以及分别用胡黄连苷Ⅱ5 mg／kg、10 mg／kg、20 mg／kg和40 mg／kg亚组。用双侧颈总动脉结扎法制作脑缺血大鼠模型，治疗组大鼠按相应时间点和剂量经腹腔注射胡黄连苷Ⅱ。用HE染色观察脑变性细胞指数（ DCI）及形态；电镜观察脑组织超微结构。免疫组化染色和Western blot法检测脑组织MMP－9蛋白水平，逆转录（ RT）－PCR检测MMP－9 mRNA水平。结果脑缺血组大鼠脑组织损害和MMP－9表达水平明显高于正常对照组（均P＜0．05）。与脑缺血组比较，治疗组的脑组织损害及DCI和MMP－9表达水平明显减轻和降低（均P＜0．05）。治疗组各亚组间两两比较，缺血1 h亚组与2 h、2．5 h亚组间脑DCI的差异有统计学意义（均P＜0．05）；缺血2 h与2．5 h亚组间的脑MMP－9阳性细胞指数，缺血1 h与1．5 h、2 h亚组，1．5 h与2．5 h亚组，2 h与2．5 h亚组间的脑MMP－9蛋白水平的差异均有统计学意义（均P＜0．05）；以及剂量5 mg／kg与20 mg／kg亚组间的脑MMP－9 mRNA水平的差异有统计学意义（均P＜0．05）。各剂量亚组间的脑DCI、MMP－9阳性细胞指数和蛋白水平的差异均无统计学意义（均P＞0．05）。结论胡黄连苷Ⅱ能抑制大鼠脑缺血后MMP－9的表达，并有显著的脑保护作用。按治疗时间窗最大化和用药剂量最小化的原则，其最佳的治疗时间窗为脑缺血后2 h内，剂量为（10～20） mg／kg。Objective To explore the effect of picrosideⅡon cerebral ischemic injury and expression of matrix metalloproteinase-9 （ MMP-9） in rats, and the selection of the best time window and dose of treatment.Methods A total of 174 Wistar rats were randomly divided into cerebral ischemia group, normal control group and treatment group.The rats in treatment group were randomly divided into four subgroups with therapeutic time window designed：1.0 h, 1.5 h, 2.0 h, 2.5 h after ischemia, and four subgroups with therapeutic drug picroside Ⅱ dose which has four levels as follows：5 mg/kg, 10 mg/kg, 20 mg/kg, 40 mg/kg body weight according to the principle of orthogonal experimental design.The forebrain ischemia models were established by bilateral common carotid artery occlusion （ BCCAO） methods.The rats in treatment group were injected intraperitoneally according to the corresponding doses designed in the orthogonal layout with picrosideⅡ.The morphology of nerve cell and the denatured cell index（ DCI） were observed by HE staining.The ultrastructure of brain tissue was observed by transmission electron microscopy （TEM）.The expression of MMP-9 protein was determined by immunohistochemical staining and Western blot method.The expression of MMP-9 mRNA was detected by reverse transcription-polymerase chain reaction （ RT-PCR）.Results The cerebral ischemic injury and the expression of MMP-9 in cerebral ischemia group rats increased significantly compared with those in normal control group （ all P〈0.05 ） , and the cerebral ischemic injury and the expression of MMP-9 in treatment group were alleviated and reduced obviously compared with cerebral ischemia group （ all P〈0.05 ） . All data from treatment group was analyzed by the least significant difference for pairwise comparisons and the results showed that there was significant deviation of DCI between ischemia 1.0 h and 2.0 h, and ischemia 1.0 h and 2.5 h（all P〈0.05）.Also there was significant deviation of the positive cell index of MMP-9

关 键 词：脑缺血 胡黄连苷Ⅱ 治疗时间窗 剂量 基质金属蛋白酶-9 

分 类 号：R743.3[医药卫生—神经病学与精神病学]