蚓激酶在缺血再灌注模型中的作用及机制研究

The role and mechanism of lumbrokinase in ischemia-reperfusion model

目的探索蚓激酶在大鼠脑缺血再灌注模型中的作用及机制。方法40只SPF级健康雄性成年大鼠采用随机数字表法分为观察组15只、模型组15只和对照组10只。观察组和模型组大鼠行大鼠脑中动脉缺血再灌注模型建模,对照组仅分离左侧颈总动脉、颈外动脉和颈内动脉。观察组每只大鼠给予6 ml/d蚓激酶溶液(约120 U蚓激酶)灌胃给药,模型组和对照组大鼠给予0.9%氯化钠注射液6 ml/d,连续给药14 d。给药结束后,处死3组大鼠,留取脑组织。处死前使用改良的Longa五级4分法对3组大鼠进行神经功能评分,使用2,3,5-三苯基四唑氯化物法染色,继而测定3组大鼠脑梗死体积。使用RT-PCR检测3组大鼠脑组织中炎症因子mRNA表达水平,使用ELISA法检测3组大鼠脑组织中炎症因子蛋白表达水平。结果观察组和模型组大鼠神经功能评分高于对照组,脑梗死体积大于对照组,观察组神经功能评分低于模型组,脑梗死体积小于模型组,差异均有统计学意义(均P<0.05)。观察组和模型组IL-1β、TNF-α、IL-6的mRNA和蛋白表达水平高于对照组,观察组上述指标表达水平低于模型组,差异均有统计学意义(均P<0.05)。模型组IL-10的mRNA和蛋白表达水平低于对照组,观察组IL-10的mRNA和蛋白表达水平高于模型组,差异均有统计学意义(均P<0.05)。结论蚓激酶在缺血再灌注大鼠模型中发挥保护作用,可能是通过抑制炎症因子表达来实施的。

Objective To explore the role and mechanism of lumbrokinase in rat cerebral ischemia-reperfusion model.Methods Forty healthy adult male SPF rats were randomly divided into observation group(n=15),model group(n=15)and control group(n=10).Rat cerebral ischemia-reperfusion model was established in observation group and model group.Only the left common carotid artery,external carotid artery and internal carotid artery were separated in the control group.Each rat in the observation group was given 6ml/d lumbrokinase solution(about 120U lumbrokinase)by gavage,while rats in the model group and control group were given the same dose of 0.9%sodium chloride injection for 14 consecutive days.After the administration,rats in 3 groups were sacrificed and brain tissues were collected.Before sacrifice,neurological function scores of rats in 3 groups were evaluated by modified Longa five-grade four-point method.Infarct volume was measured by using 2,3,5-triphenyltetrazolium chloride staining.RT-PCR was used to detect the mRNA expression levels of inflammatory factors in brain tissues of rats in 3 groups.ELISA was used to detect the protein expression levels of inflammatory factors in brain tissues of rats in 3 groups.Results The neurological function scores and infarct volume of rats in observation group and model group were higher than those in control group.The neurological function scores were lower and infarct volume was smaller in observation group than in model group,the differences were statistically significant(all P<0.05).The mRNA and protein expression levels of IL-1β,TNF-αand IL-6 in observation group and model group were higher than those in control group.The above indicators in observation group were lower than those in model group,the differences were statistically significant(all P<0.05).The mRNA and protein expression levels of IL-10 in model group were lower than those in control group.The mRNA and protein expression levels of IL-10 in observation group were higher than those in model group,the differences were statistically significant(all P<0.05).Conclusion Lumbrokinase plays a protective role in ischemia-reperfusion rat model,which may be achieved by inhibiting the expression of inflammatory factors.