针刺“百会”“太阳”改善局灶性脑缺血脑微血管内皮细胞功能的动态观察

目的:在分子水平探讨并阐明脑缺血损伤的炎性机理和针刺对局灶性脑缺血模型脑微血管内皮细胞功能的动态影响。方法:老龄雄性Wistar大鼠80只,随机分为缺血模型对照组、针刺治疗组各32只,假手术组、正常对照组各8只,前二组又分为造模后针刺1、3、51、0 d 4个时间段组,每时间段各8只。选取大鼠“百会”“太阳”穴进行手捻针治疗。采用大脑中动脉线栓阻塞法建立大鼠局灶性脑缺血模型;用免疫组化SABC法检测脑缺血后治疗1、3、51、0 d内皮素(ET-1)、FⅧ因子相关抗原和细胞间粘附分子(ICAM-1)的表达。结果:针刺不同疗程治疗,各组大鼠在治疗前后的神经症状行为评定分析有显著性差异(P<0.05)。模型组ET-1与ICAM-1均脑缺血后1 d即出现表达,各个时间点相比具有显著性差异(P<0.01);针刺治疗随时间的延长可明显降低其表达(P<0.05,0.01)。FⅧ相关抗原在微血管内皮细胞中的表达在缺血后3 d开始增加(P<0.05),并持续上升,各个时间点的差异具有统计学意义(P<0.01);针刺治疗随时间的延长可使其表达继续升高(P<0.05,0.01)。结论:针刺对脑缺血状态下海马CA3区多层面进行良性调节,并且这种良性机制随着干预时间的选择(早期/6 h)和疗程的适度延长(5 d以上),具有明显的累加效应。

Effects of Acupuncture of "Baihui" (GV 20) and "Taiyang" (EX-HN 5) on Functions of Vascular Endothelial Cells in Cerebral Ischemia Injury Rats

Objective:To study the mechanism of acupuncture of "Baihui"(GV 20) and "Taiyang"(EX-HN 5) in improving cerebral ischemia(CI) injury and vascular endothelial cellular function.Methods: Eighty old male Wistar rats were randomized into control(n=8),model(n=32),acupuncture(n=32),sham-operation(n=8) groups.Model and acupuncture groups were further evenly and respectively divided into(1 d),(3 d),(5 d) and(10 d) subgroups.CI model was established by using middle cerebral artery occlusion(MCAO) method."Baihui"(GV 20) and "Taiyang"(EX-HN 5) were punctured with filiform needles which were twirled rapidly at a frequency of about 200 revolutions/min for(1 min)(1 time/10 min),and retained for 30 min.The treatment was given once daily,continuously for 1,3,5 and 10 days respectively in different subgroups.Endothelin(ET)-1,intercellular adhesion molecule (ICAM)-1,and factor Ⅷ related antigen(FⅧR-Ag) of CA_3 area tissue of the hippocampus were assayed with immunohistochemical method(SABC).Results: The CI rats' behavioral symptoms of(5 d) and(10 d) groups were improved significantly in comparison with the corresponding subgroups of model group (P<0.05,0.01).Compared with(1 d),(3 d),(5 d) and(10 d) subgroups of control group,the total area and integral optical density(IOD) values of ICAM-1,ET1 and FⅧR-Ag expression in CA_3 of hippocampus all increased significantly in the 4 subgroups of model group(P<0.01).Compared with 4 subgroups of model group,the total area values of acupuncture group,ICAM-1 expression in the 4 subgroups,ET-1 in(5 d) and(10 d) subgroups decreased significantly,and FⅧR-Ag expression in(3 d),(5 d) and(10) d subgroups increased;IOD values of ICAM-1 in(3 d),(5 d) and(10 d) subgroups,ET-1 in(5 d) and(10 d) subgroups decreased markedly,and FⅧR-Ag in(3 d),(5 d) and(10 d) subgroupsall increased considerably(P<0.05,0.01).Conclusion: Acupuncture of "Baihui"(GV 20) and "Taiyang"(EX-HN 5) can improve CI rats' behavior and regulate the expression of vascular endothelial ICAM-1,ET-1 and FⅧR-Ag in CA_3 of hippocampus and has accumulative effects.