PSD-95抑制剂ZL006对新生大鼠缺氧缺血性脑损伤的作用探讨

The effect of PSD-95 inhibitor ZL006 on hypoxic-ischemic brain damage in neonatal rats

目的探讨PSD-95抑制剂ZL006对新生大鼠缺氧缺血性脑损伤(hypoxic-ischemic brain damage,HIBD)的作用及其发病机制。方法选取7日龄健康Wistar新生大鼠(80只),随机将其分为正常对照组(20只)、假手术组(20只)、手术组(即HIBD模型组40只)。将手术组又随机分为ZL006治疗组(采用ZL00610 mg/kg进行腹腔注射,20只)与非治疗组(20只)。于手术后第1天、第7天随机选取各组新生大鼠,通过氯化三苯基四氮唑(triphenyl tetrazolium chloride)染色法观察脑组织梗死程度;通过蛋白印迹法监测损伤区域脑组织PSD-95蛋白表达;采用ELISA评价ZL006对HIBD大鼠氧化应激反应及抗氧化酶的影响。结果(1)术后第1天,非治疗组大鼠脑损伤最严重,ZL006治疗组脑梗死减少,术后第7天手术组仍可见少许梗死灶。(2)术后第1天,手术组PSD-95蛋白表达量显著高于正常对照组及假手术组(P<0.01),ZL006治疗组低于非治疗组(P<0.05);术后第7天,各组间PSD-95蛋白表达量差异无统计学意义(P>0.05)。(3)术后第1天,手术组4-羟基壬烯醛表达量显著高于正常对照组及假手术组(P<0.01),且非治疗组高于ZL006治疗组(P<0.05);术后第7天各组间4-羟基壬烯醛表达量差异无统计学意义(P>0.05)。(4)术后第1天,手术组中超氧化物歧化酶表达量显著低于正常对照组及假手术组(P<0.01),且非治疗组低于ZL006治疗组(P<0.05);术后第7天各组间超氧化物歧化酶表达量差异无统计学意义(P>0.05)。结论PSD-95可能参与了HIBD的早期发病过程,其抑制剂ZL006对新生大鼠HIBD可能存在神经保护作用。

Objective To explore the effect of PSD-95 inhibitor ZL006 in neonatal rats with hypoxic-ischemic brain damage(HIBD)and its potential mechanism.Methods The seven-day-old healthy Wistar rats(n=80)were randomly divided into control group(n=20),sham operation group(n=20)and operation group(HIBD model group,n=40).The operation group was randomly divided into ZL006,treatment group(intraperitoneal injection of ZL006,10 mg/kg,n=20)and non-treatment group(n=20).The neonatal rats of each group were randomly selected on the 1st and 7th day after operation,and the degree of cerebral infarction was observed by triphenyl tetrazolium chloride staining.The protein expression level of brain tissue in the injured area was examed by Western blot,and the effects of ZL006 on oxidative stress and antioxidant enzymes in rats with HIBD were evaluated by ELISA.Results(1)On the first day after operation,the brain injury was the most serious in the non-treatment group,and the cerebral infarction decreased in the ZL006 treatment group.On the 7th day after operation,a little infarction could be seen in the operation group,but there was no significant difference among other three groups.(2)On the first day after operation,the expression of PSD-95 protein in the operation group was significantly higher than that in the control group and sham operation group(P<0.01).There was significant difference in the expression of PSD-95 protein between the ZL006 treatment group and the non-treatment group(P<0.05).On the 7th day after operation,there was no significant difference in the expression of PSD-95 protein among three groups.(3)On the first day after operation,the expression of 4-hydroxynonenal in the operation group was significantly higher than that in the control group and sham operation group(P<0.01),and that in the non-treatment group was higher than that in the ZL006 treatment group(P<0.05).On the 7th day after operation,there was no significant difference in the expression of 4-hydroxynonenal among three groups.(4)On the first day after operation,the expression of superoxide dismutase in the operation group was significantly lower than that in the control group and sham operation group(P<0.01),and that in the non-treatment group was lower than that in the ZL006 treatment group(P<0.05).There was no significant difference in the expression level of superoxide dismutase among three groups on the 7th day after operation.Conclusion It is suggested that PSD-95 may be involved in the early pathogenesis of HIBD,and ZL006 may have neuroprotective effect on HIBD in newborn rats.