摘要
目的:通过观察眼针对CIRI大鼠脑组织中IL-1β、IL-18含量和P2X7R、NLRP3、pro-caspase-1、ASC、caspase-1表达水平的影响,探讨眼针通过调控细胞焦亡途径发挥对CIRI大鼠抗炎作用的机制。方法:SD大鼠85只,随机分为假手术组(16只)和造模组(69只),采用改良的线栓法对造模组大鼠进行CIRI模型复制,假手术组大鼠采用相同术式进行手术,但不进行大脑中动脉栓塞。术后以神经功能缺损评分和脑组织TTC染色对CIRI大鼠进行模型质量评价,将造模成功的大鼠再次随机均分为:模型对照组、眼针组、抑制剂组。然后对于眼针组大鼠行眼针干预,干预时机分别为:再灌注即刻及再灌注8、16、24 h,共计进行4次干预。抑制剂组大鼠一次性经腹腔注射格列本脲,500 mg/kg。假手术组和模型对照组大鼠不作处理,正常饲养至实验结束。末次针刺后,对各组大鼠进行神经功能缺损评分,然后采用过量麻醉法处死大鼠,用免疫组织化学法检测、Western blot法检测脑组织中P2X7R、NLRP3、pro-caspase-1、ASC、caspase-1的表达水平;ELISA法检测脑缺血半暗带组织中IL-1β、IL-18的含量。结果:与假手术组比较,模型对照组大鼠脑缺血半暗带组织中IL-1β、IL-18含量显著升高,P2X7R、NLRP3、pro-caspase-1、ASC、caspase-1的表达水平显著上调,差异有统计学意义(P<0.01);与模型对照组比较,眼针组和抑制剂组大鼠脑缺血半暗带组织中IL-1β、IL-18含量显著降低,NLRP3、pro-caspase-1、ASC、caspase-1的表达水平显著下调,差异有统计学意义(P<0.01)。结论:CIRI损伤可能引起缺血半暗带脑组织中细胞焦亡的发生,同时脑组织中细胞因子IL-1β、IL-18高表达,P2X7R、NLRP3、pro-caspase-1、ASC、caspase-1的表达高于假手术组;而眼针干预能够抑制IL-1β、IL-18及P2X7R、NLRP3、pro-caspase-1、ASC、caspase-1的表达,进而调控细胞焦亡的发生而发挥抗炎作用。
Objective:To observe the content of IL-1β,IL-18 content,P2 X7 R,NLRP3,pro-caspase-1,ASC and caspase-1 expression in CIRI rats,and to explore the anti-inflammatory mechanism of eye-acupuncture on CIRI rats.Methods:Eighty-five SD rats were randomly divided into the Sham-operated group(16 rats) and the operation group(69 rats).The modified suture method was used to replicate the CIRI model in rats of the Operation group.After successful modeling,the operation rats were randomly divided into model control group,eye-acupuncture group and inhibitor group.Then the rats in the eye-acupuncture group were intervened by eye-acupuncture at the time of immediate reperfusion and 8,16 and 24 hours of reperfusion.In the inhibitor group,glibenclamide(500 mg/kg) was injected intraperitoneally.The rats in the Sham-operated group and the model control group were fed normally until the end of the experiment.After the last acupuncture,the rats in each group were scored for neurological deficit,and then the rats were killed by excessive anesthesia,and the expression levels of P2 X7 R,NLRP3,pro-caspase-1,ASC and caspase-1 in brain tissues were detected by the methods of immunohistochemistry and Western blot.ELISA was used to detect the contents of IL-1β and IL-18 in cerebral ischemic penumbra tissues.Results:Compared with the model control group,the contents of IL-1β and IL-18 in cerebral ischemic penumbra tissue of rats in eye-acupuncture group and inhibitor group decreased significantly,while the expressions of NLRP3,pro-caspase-1,ASC and caspase-1 decreased significantly(P<0.01).Conclusion:In CIRI rats,CIRI injury may cause apoptosis in ischemic penumbra,and the expression of apoptosis related protein in brain tissue is higher than that in sham-operated group.Eye-acupuncture therapy can inhibit the occurrence of pyroptosis by reducing the expression of apoptosis related cytokines and and play an anti-inflammatory role.
作者
刘昱麟
马贤德
宋采秋
徐畅
张双双
王哲
LIU Yu-Lin;MA Xian-De;SONG Cai-Qiu;XU Chang;ZHANG Shuang-Shuang;WANG Zhe(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2019年第24期2964-2970,共7页
Chinese Journal of Immunology
基金
国家自然科学基金面上项目(No.8167150734)
国家自然科学基金青年基金(No.81703993)
辽宁省自然科学基金(No.201602507)
辽宁省教育厅项目(No.L201714)
关键词
眼针
缺血性脑中风
细胞焦亡
Eye-acupuncture
Ischemic stroke
Pyroptosis
