康益胶囊对缺血再灌注脑水肿大鼠脑组织AQP4、MMP-9蛋白的影响

Effect of Kangyi capsule on AQP-4 and MMP-9 protein in brain tissue of rats with cerebral ischemia-reperfusion edema

目的研究康益胶囊对缺血再灌注脑水肿大鼠脑组织水通道蛋白(AQP)4、基质金属蛋白酶(MMP)-9的影响。方法选用清洁级健康雄性SD大鼠72只,采用数字表法随机分为假手术组、模型组、康益胶囊低剂量组(270 mg/kg)、中剂量组(540 mg/kg)、高剂量组(1080 mg/kg)、银杏叶片组(680 mg/kg),每组12只,模型组和用药组采用线栓法制作大脑中动脉栓塞(MCAO)大鼠模型,缺血1 h行再灌注,假手术组行假手术。术后连续灌胃给药14 d,给药期间定期观察各组脑损伤后神经功能症状,14 d后断头取脑,检测脑组织含水量、观察2,3,5-三苯基氯化四氮唑(TTC)染色后脑梗死面积、苏木素-伊红(HE)染色观察损伤脑区的病理损伤,Western印迹法检测AQP4、MMP-9蛋白表达水平。结果与假手术组相比,模型组神经功能缺损症状明显、脑组织含水量显著增加(均P<0.01);与模型组相比,康益胶囊低、中、高及银杏叶片组均能显著改善脑缺血再灌注大鼠神经功能缺损症状、显著降低脑组织含水量(均P<0.01);TTC染色结果表明,与假手术组相比,模型组脑梗死面积显著(P<0.01);与模型组相比,康益胶囊低、中、高及银杏叶片组脑梗死面积明显减小(均P<0.01);与银杏叶片组相比,康益胶囊高剂量组减少脑梗死面积明显(P<0.05);Western印迹法结果显示,与模型组相比,康益胶囊可显著降低脑缺血再灌注大鼠缺血侧脑组织中AOP-4及MMP-9蛋白表达水平(均P<0.01)。结论康益胶囊可降低缺血再灌注脑水肿大鼠脑组织中AQP4、MMP-9蛋白表达而发挥脑保护作用。

Objective To study the effect of Kangyi capsule on aquaporin(AQP)-4 and matrix metalloproteinase(MMP)-9 in brain tissue of rats with cerebral ischemia-reperfusion edema.Methods 72 clean healthy male SD rats were divided into sham operation group,model group,Kangyi capsule low dose group(270 mg/kg),medium dose group(540 mg/kg),high dose group(1080 mg/kg)and ginkgo biloba tablet group(680 mg/kg)by random number table method,with 12 rats in each group.Middle cerebral artery occlusion(MCAO)rat model was made by thread embolization method in model group and treatment group,reperfusion was performed at 1 h after ischemia.Sham operation was performed in the sham operation group.After 14 d of continuous intragastric administration,the neurological symptoms after brain injury in each group were observed regularly during administration.After 14 d,the brain was decapitated to take brain.The water content of brain tissue was measured,the cerebral infarction area was observed after 2,3,5-triphenyltetrazolium chloride(TTC)staining,the pathological injury of the injured brain area was observed by hematoxylin-eosin(HE)staining,and the protein expression levels of AQP4 and MMP-9 were detected by Western blotting.Results Compared with the sham operation group,the symptoms of neurological deficit and the water content of brain tissue in the model group were significantly increased(all P<0.01).Compared with the model group,Kangyi capsule low,medium,high and ginkgo biloba tablet groups could significantly improve the symptoms of neurological deficit and significantly reduce the water content of brain tissue in rats with cerebral ischemia-reperfusion(all P<0.01).The results of TTC staining showed that compared with the sham operation group,the cerebral infarction area of the model group was significantly lower(P<0.01).Compared with the model group,the cerebral infarction area of Kangyi capsule low,middle,high and ginkgo biloba tablet groups were significantly decreased(all P<0.01).Compared with the ginkgo biloba tablet group,the cerebral infarction area of the high dose of Kangyi capsule group were significantly decreased(P<0.05).The results of Western blotting showed that compared with the model group,Kangyi capsule could significantly reduce the protein expression levels of AOP-4 and MMP-9 in the ischemic brain tissue of rats with cerebral ischemia-reperfusion(all P<0.01).Conclusions Kangyi capsule could protect the brain by reducing the protein expression of AQP4 and MMP-9 in brain tissue of rats with cerebral ischemia-reperfusion edema.