针刺预处理对脑梗死大鼠胆碱能抗炎通路的影响研究

摘要

Objective To explore the impact of acupuncture preconditioning on cholinergic anti - inflammatory pathway in rats with cerebral infarction. Methods A total of 40 adult male SD rats were randomly divided into groups A,B, C,D,E,each of 8 cases. Rats of A group were given normal feeding,rats of B group were given sham-operation( cut the skin and exposed carotid general artery,external carotid artery,internal carotid artery,did not insert line-lock,then suture the skin),rats of C group were prepared for cerebral infarction model according to modified Zea Longa method,rats of D group were given acupuncture preconditioning 7 days before modeling,rats of E group were given acupuncture preconditioning and methyllycaconitine 7 days before modeling. All of the rats were killed 3 days after modeling,and neurological score was evaluated by Longar 5 -grade method before killing. Fresh parietal lobe cortex were collected and fixed by formaldehyde,and TUNEL method was used to detect the cell apoptosis;other brain tissues were kept by liquid nitrogen,and ELISA was used to detect the TNF-α and IL-1β levels,real -time fluorescent quantitative PCR was used to detect the expression of α7nAChR mRNA. Results TUNEL positive cells of groups C,D,E were more than those of groups A and B,respectively;TNF-αand IL-1βlevels of groups C,D,E were higher than those of groups A and B,respectively;while the expression of α7nAChR mRNA of groups C,E were lower than those of groups A and B,respectively(P<0. 05). TUNEL positive cells of D group were less than those of groups C and E,respectively;neurological score,TNF -α and IL -1β levels of D group were lower than those of groups C and E,respectively;while the expression of α7nAChR mRNA of D group were higher than those of groups C and E, respectively( P<0. 05 ). Conclusion Acupuncture preconditioning can improve neurological function in rats with cerebral infarction,inhibit neuronal apoptosis and reduce inflammatory cytokines levels,raise the expression of 7nAChR mRNA.%目的：探讨针刺预处理对脑梗死大鼠胆碱能抗炎通路（ CAP）的影响。方法选择40只健康成年SD雄性大鼠，采用随机数字表法将大鼠分为正常组（A组）、假手术组（B组）、脑梗死组（C组）、针刺预处理组（D组）和针刺预处理+甲基牛扁亭（ MLA）组（ E组），各8只。A组大鼠不做任何处理；B组大鼠只切开皮肤，暴露颈总动脉、颈外动脉、颈内动脉，不插入线栓，术后缝合皮肤；C组大鼠采用改良Zea Longa方法制备脑梗死模型；D组于造模前7 d开始针刺治疗；E组大鼠于针刺同时腹腔注射MLA，5 mg/kg，1次/d，连续治疗7 d。各组大鼠于造模成功后同步饲养，自由饮食，3 d后处死。于处死前采用Longar 5分制评分法进行神经功能评分；取处死大鼠新鲜脑组织顶叶皮质采用甲醛固定，采用 TUNEL 法检测细胞凋亡情况；剩余脑组织采用液氮保存，采用酶联免疫吸附试验（ELISA）检测TNF-α、IL-1β水平，采用实时定量荧光PCR检测α7烟碱型乙酰胆碱受体（α7nAChR）mRNA表达情况。结果 C组、D组、E组大鼠TUNEL阳性细胞多于A组和B组，脑组织中TNF-α、IL-1β水平高于A组和B组（P<0.05）；C组和E组大鼠α7nAChR mRNA相对表达量低于A组和B组（P<0.05）；D组大鼠TUNEL阳性细胞少于C组和E组，神经功能评分低于C组和E组，脑组织中TNF-α、IL-1β水平低于C组和E组，α7nAChR mRNA相对表达量高于C组和E组（P<0.05）。结论针刺预处理可以改善脑梗死大鼠神经功能，抑制神经元凋亡，降低炎性细胞因子水平，上调α7nAChR mRNA的表达。