参附注射液对大鼠脑缺血再灌注损伤后MMP-9和TIMP-1表达的影响

目的 观察参附注射液对大鼠脑缺血再灌注后基质金属蛋白酶-9(MMP-9)及基质金属蛋白酶抑制剂-1(TIMP-1)表达的影响,探讨参附注射液的脑保护机制. 方法 Wistar大鼠90只按随机数字表法分为假手术组、参附治疗组、模型组,每组30只.后2组大鼠采用线栓法经左侧颈外一颈内动脉插线建立脑缺血再灌注模型,并分别于再灌注时静脉注射等量参附注射液和生理盐水.每组根据缺血时间分为0.5、1、1.5h3个亚组,每亚组10只,再灌注24h后将大鼠处死以干湿重法检测缺血再灌注侧脑组织含水量,Western blotting检测脑组织中MMP-9和TIMP-1的表达.结果 模型组与参附治疗组在脑缺血不同时间脑组织含水量、MMP-9、TIMP-1的表达均高于假手术组,并且随着缺血时间的增加,脑组织含水量和MMP-9的表达增加,差异有统计学意义(P＜0.05)；与模型组比较,参附治疗组在不同脑缺血时间脑组织的含水量、MMP-9的表达较低,而TIMP-1蛋白的表达较高,差异均有统计学意义(P＜0.05). 结论 参附注射液可能通过调节MMP-9、TIMP-1的表达发挥抗脑水肿的作用.

Effect of Shenfu injection on expressions of matrix metalloproteinase-9 and tissue inhibitors of metalloproteinase-1 in rats after cerebral ischemia-reperfnsion injury

Objective To investigate the effect of Shenfu injection on the expressions of matrix metalloproteinase-9 (MMP-9) and tissue inhibitors of metalloproteinase-1 (TIMP-1) in rats after cerebral ischemia-reperfusion injury,and explore the brain protection mechanism of Shenfu injection. Methods Ninety adult healthy male Wister rats were chosen and randomly divided into sham-operated group,Shenfu injection treatment group and vehicle group (n=30); animal models of middle cerebral artery occlusion/reperfusion in the later 2 groups were established by a filament method in the left external-internal carotid artery.Rats in the Shenfu injection treatment group were injected with 8 mg/kg Shenfu injection during the reperfusion while those in the vehicle group with physiological saline.These rats in each group were divided into 3 sub-groups (n=10) according to the ischemia time (0.5,1 and 1.5 h after the model establishment). Twenty-four h after reperfusion, dry-weight method was employed to detect the content of brain tissues on the side received ischemia-reperfusion; and Western blotting was employed to observe the expressions of MMP-9 and TIMP-1 in the brain tissues. Results The water content of those brain tissues and the expressions of MMP-9 and TIMP-1 in the Shenfu injection treatment group and vehicle group at each time point significantly increased as compared with those in the sham-operated group (P＜0.05); the longer the ischemic time,the heavier the water content of the brain tissues and the higher the expressions of MMP-9 and TIMP-1. As compared with those in the vehicle group, the water content of the brain tissues and the expression of MMP-9 in the Shenfu injection treatment group obviously decreased,while the expression of TIMP-1 in the Shenfu injection treatment group significantly increased at each time point (P＜0.05). Conclusion Shenfu injection has the function against cerebral edema by accommodating the expressions of MMP-9 and TIMP-1.