摘要
目的:探究黄芪赤风汤对大脑中动脉栓塞(MCAO)大鼠模型的治疗作用及机制。方法:将90只SPF级雄性大鼠随机分为假手术组,模型组,黄芪赤风汤高、中、低剂量组(中药低、中、高剂量组)(8.10、4.05、2.025 g·kg^(-1)),脑心通组(脑心通,0.32 g·kg^(-1)),建立MCAO大鼠模型,给予黄芪赤风汤药液进行干预,对各组大鼠进行神经功能学评分;大鼠脑组织采用2,3,5-氯化三苯四氮唑(TTC)染色,计算脑梗死面积;采用酶联免疫吸附测定法(ELISA)检测各组大鼠血清白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、基质金属蛋白酶-9(MMP-9)、血管内皮生长因子(VEGF)和血管内皮生长因子受体2(VEGFR2)含量;采用苏木素-伊红(HE)染色观察各组大鼠脑组织病理变化;蛋白免疫印迹法(Western blot)检测大鼠脑组织闭锁小带蛋白-1(ZO-1)、紧密连接蛋白-5(Claudin-5)、P-糖蛋白(P-gp)和多药耐药蛋白1(MRP1)的表达。结果:与假手术组比较,模型组大鼠神经功能评分、脑梗死率显著增加(P<0.01),血清IL-6、IL-1β和MMP-9含量显著升高(P<0.01),脑组织ZO-1,Claudin-5蛋白表达水平显著降低(P<0.01),P-gp,MRP1蛋白表达水平显著升高(P<0.01);与模型组比较,各给药组大鼠14 d神经功能评分明显降低(P<0.05,P<0.01),且有效改善脑组织病理变化,显著降低脑梗死率(P<0.01),血清IL-6、IL-1β、MMP-9表达水平明显降低(P<0.05,P<0.01),VEGFR2含量显著增加(P<0.01),中药高、中剂量组及脑心通组VEGF含量明星增加(P<0.05,P<0.01),中药低剂量组虽有下降趋势,但差异无统计学意义,中药高剂量组和脑心通组脑组织ZO-1、Claudin-5蛋白表达水平明显升高(P<0.05,P<0.01),P-gp、MRP1蛋白表达水平明显降低(P<0.05)。结论:黄芪赤风汤可以通过改善脑组织病理变化,减少炎症反应、促进血管生成、调节血脑屏障(BBB)功能,从而发挥治疗脑梗死作用。
Objective:To explore the therapeutic effect and mechanism of Huangqi Chifengtang on middle cerebral artery embolism(MCAO)rat model.Method:The 90 SPF male rats were randomly divided into sham operation group,model group,high,medium and low dose groups of Huangqi Chifengtang(8.10,4.05,2.025 g·kg^(-1))and positive drug group(Naoxintong 0.32 g·kg^(-1)).MCAO rat model was established and intervened with Huangqi Chifengtang,and the neurological scores of each group were scored.The area of cerebral infarction was calculated by 2,3,5-triphenyltetrazolium chloride(TTC)staining.Serum interleukin-6(IL-6)and interleukin-1β(IL-1β)were detected by enzyme-linked immunosorbent assay(ELISA),The contents of matrix metalloproteinase-9(MMP-9),vascular endothelial growth factor(VEGF)and vascular endothelial growth factor receptor 2(VEGFR2),the pathological changes of brain tissue in each group were observed by hematoxylin eosin(HE)staining.Western blot was used to detect zonule atresia protein-1(ZO-1),tight junction protein-5(Claudin-5)and P-glycoprotein(P-gp)and multidrug resistance protein 1(MRP1).Result:Compared with the sham operation group,the neurological function score and cerebral infarction rate of the model group were significantly increased(P<0.01),and the levels of IL-6,IL-1βand MMP-9 in serum were significantly increased(P<0.01),the levels of ZO-1 and Claudin-5 protein expression decreased significantly(P<0.01),and the levels of P-gp and MRP1 protein expression increased significantly(P<0.01).Compared with the model group,the neurological function score of rats in each administration group decreased significantly at 14 days(P<0.05,P<0.01),the pathological changes of brain tissue effectively improved,the rate of cerebral infarction significantly reduced(P<0.01),and the expression level of IL-6,IL-1βand MMP-9 in serum decreased significantly(P<0.05,P<0.01),the content of VEGFR2 increased significantly(P<0.01),and the content of VEGF increased significantly in high,medium dose and positive drug groups(P<0.05,P<0.01).Although it decreased in low dose group,there was no significant difference.The levels of ZO-1 and Claudin-5 protein expression in brain tissue of high dose group and positive drug group increased significantly(P<0.05,P<0.01),the level of MRP1 and P-gp protein expression decreased significantly(P<0.05).Conclusion:Huangqi Chifengtang can play a therapeutic role in rats with cerebral infarction by improving the pathological changes of brain tissue,reducing inflammatory reaction,promote angiogenesis and regulating the function of blood-brain barrier(BBB).
作者
刘舒毓
王秋月
刘树民
LIU Shu-yu;WANG Qiu-yue;LIU Shu-min(Heilongjiang University of Chinese Medicine,Institute of Traditional Medicine,Harbin 150040,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2022年第6期26-33,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
黑龙江省自然科学基金项目(ZD2020H006)
黑龙江中医药大学优秀创新人才(领军人才)项目(2018RCL13)。
关键词
黄芪赤风汤
脑梗死
炎症
血管生成
血脑屏障
紧密连接蛋白
Huangqi Chifengtang
cerebral infarction
inflammation
angiogenesis
blood brain barrier
tight junctional protein
作者简介
第一作者:刘舒毓,在读硕士,从事中药药性理论及药效物质基础研究,E-mail:1366076152@qq.com;通信作者:刘树民,博士,教授,博士生导师,从事中药药性理论及药效物质基础研究,E-mail:keji-liu@163.com。
