电针对脑缺血-再灌注损伤大鼠运动功能的改善作用及机制研究
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摘要
目的探讨电针对缺血性脑损伤后GRP78和caspase12的影响及对大鼠肢体运动功能修复的影响及机制。方法本实验使用改良线栓法制备大鼠脑缺血-再灌注模型(MCAO),对"百会"和"水沟"穴电针刺激,电针干预时间30min。SD大鼠随机分为对照组(sham组)、脑缺血-再灌注(3h、6h、12h、24h、72h、120h)组(I/R组)和再灌注时间点+电针组(I/R+EA组),每组5只,转棒式疲劳仪实验检测大鼠肢体运动功能的修复状况,RT-PCR检测caspase-12 mRNA的表达,免疫荧光实验检测GRP78蛋白的表达。结果 I/R+EA组(7d和14d)大鼠在转棒上停留时间与对应I/R组相比显著延长,差异有统计学意义(P<0.05),显示电针干预能促进大鼠运动功能的修复;I/R+EA组(3h、6h、12h、24h、72h和120h)缺血侧脑组织caspase12 mRNA的表达与对应I/R组相比显著降低,差异有统计学意义(P<0.05),I/R+EA组(3h、6h、12h和24h)GRP78阳性细胞数与对应I/R组相比显著减少,差异有统计学意义(P<0.05)。结论电针干预能促进缺血性脑损伤大鼠肢体运动功能修复可能与阻断内质网介导的凋亡通路有关。
Objective To investigate the effect and possible mechanism of electroacupuncture on cerebral cellular apoptosis and neural function repair in rats with ischemic stroke. Methods Simple random sampling divided 65 rats into control group( sham),groups of cerebral ischemia/reperfusion( I/R) for 3 h,6 h,12 h,24 h,72 h,120 hand groups of cerebral I/R for 3 h,6 h,12 h,24 h,72 h,120 h and acupuncture intervention( I/R + EA). Each group has 5 rats. We used thread bolt method to make model of cerebral ischemia reperfusion in rats( MCAO). Then we stimulated on "baihui"and "shuigou"acupoint. The intervention time of electroacupuncture was 30 min. The repair status of limb movement function in rats was detected by rotating-fatigue instrument,and the expression of caspase-12 mRNA was detected by RT-PCR. The expression of GRP78 protein was detected by immunofluorescence experiment. Results The duration of staying on the rotating bar in the I/R + EA group( 7 d and 14 d) was significantly longer than that in the corresponding I/R group,and the difference was statistically significant( P < 0. 05),which indicated that the electro-acupuncture intervention could promote the repair of motor function in the rats. The mRNA expression of caspase12 in ischemic lateral brain tissue of the I/R + EA group( 3 h,6 h,12 h,24 h,72 h and 120 h) was significantly lower than that of the corresponding I/R group( P < 0. 05). The number of GR P78-positive cells in the I/R + EA group( 3 h,6 h,12 h and 24 h)was significantly reduced compared with the corresponding I/R group,and the difference was statistically significant( P <0. 05). Conclusion Acupuncture intervention may prevent cerebral cellular apoptosis and promote the repair of body movement function after cerebral I/R injury by blocking apoptotic pathway mediated by endoplasmic reticulum and inflammatory factor.
Objective To investigate the effect and possible mechanism of electroacupuncture on cerebral cellular apoptosis and neural function repair in rats with ischemic stroke. Methods Simple random sampling divided 65 rats into control group( sham),groups of cerebral ischemia/reperfusion( I/R) for 3 h,6 h,12 h,24 h,72 h,120 hand groups of cerebral I/R for 3 h,6 h,12 h,24 h,72 h,120 h and acupuncture intervention( I/R + EA). Each group has 5 rats. We used thread bolt method to make model of cerebral ischemia reperfusion in rats( MCAO). Then we stimulated on "baihui"and "shuigou"acupoint. The intervention time of electroacupuncture was 30 min. The repair status of limb movement function in rats was detected by rotating-fatigue instrument,and the expression of caspase-12 mRNA was detected by RT-PCR. The expression of GRP78 protein was detected by immunofluorescence experiment. Results The duration of staying on the rotating bar in the I/R + EA group( 7 d and 14 d) was significantly longer than that in the corresponding I/R group,and the difference was statistically significant( P < 0. 05),which indicated that the electro-acupuncture intervention could promote the repair of motor function in the rats. The mRNA expression of caspase12 in ischemic lateral brain tissue of the I/R + EA group( 3 h,6 h,12 h,24 h,72 h and 120 h) was significantly lower than that of the corresponding I/R group( P < 0. 05). The number of GR P78-positive cells in the I/R + EA group( 3 h,6 h,12 h and 24 h)was significantly reduced compared with the corresponding I/R group,and the difference was statistically significant( P <0. 05). Conclusion Acupuncture intervention may prevent cerebral cellular apoptosis and promote the repair of body movement function after cerebral I/R injury by blocking apoptotic pathway mediated by endoplasmic reticulum and inflammatory factor.
引文
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[13]杜秋国.丹参预处理对大鼠肝缺血再灌注时葡萄糖调节蛋白78/94表达的影响[D].广州:南方医科大学,2013.
[14]宋小燕,赵永波,周晓琳,等.大鼠脑缺血再灌注后GRP78和GADD153的表达变化研究[J].中风与神经疾病杂志,2008,25(02):139-141.
[15]张雨平,赵聪敏,王丽雁,等.Caspase-3抑制剂抗新生鼠缺氧缺血脑损伤作用的观察[J].第三军医大学学报,2006,28(20):2068-2070.
[16]黄丽.体外扩增的调节性T细胞静脉输注移植对脑梗死的保护作用[D].泰安:泰山医学院,2013.
[2]仇健,武菲,王春梅,等.大鼠脑缺血再灌注后GRP78、CHOP表达变化[J].安徽医科大学学报,2018,53(4):497-501.
[3]樊永梅,王如蜜,张长杰,等.高频电疗对大鼠脑缺血再灌注损伤后caspase-3和海马神经细胞超微结构的影响[J].中南大学学报(医学版),2017,42(1):21-25.
[4]沈菁,雷晓明,宋洋,等.电针内关、百会穴对脑缺血/再灌注大鼠GRP78和Caspase-12基因表达的影响[J].湖南中医药大学学报,2016,36(2):54-57.
[5]戴伟娟,朱凡河,张国安,等.前列腺素E1对大鼠全脑缺血-再灌注损伤细胞凋亡及Caspase-3表达的影响[J].济宁医学院学报,2017,40(2):98-102.
[6]宋张杰.脑脉通对老龄大鼠脑缺血/再灌注脑微血管生成促进作用机制[D].郑州:河南中医药大学;2011.
[7]胡蔚婧,王华,吴松,等.针刺预处理不同配穴对延缓运动性疲劳及心脏功能调节的影响[J].湖北中医药大学学报,2018,20(3):5-8.
[8]李蓉,刘明华,张潇,等.橄榄醽的药理实验研究[J].时珍国医国药,2008,19(10):2466-2467.
[9]王海英.电针对大鼠局灶性脑缺血再灌注后脑细胞凋亡的影响及其机制的研究[D].泰安:泰山医学院,2008.
[10]田园,李运鹏,韩若东.川芎嗪对大鼠脑缺血再灌注后细胞凋亡及caspase 12表达的影响[J].临床和实验医学杂志,2014,13(1):16-19.
[11]陈露.电针对胎窘新生鼠脑Neuro D表达的影响[D].福州:福建医科大学,2015.
[12]黄秀景,高春霖,吕国义.丁苯酞氯化钠注射液预处理对局灶性脑缺血再灌注损伤大鼠GRP78及caspase-12表达的影响[J].天津医科大学学报,2013,19(3):186-188.
[13]杜秋国.丹参预处理对大鼠肝缺血再灌注时葡萄糖调节蛋白78/94表达的影响[D].广州:南方医科大学,2013.
[14]宋小燕,赵永波,周晓琳,等.大鼠脑缺血再灌注后GRP78和GADD153的表达变化研究[J].中风与神经疾病杂志,2008,25(02):139-141.
[15]张雨平,赵聪敏,王丽雁,等.Caspase-3抑制剂抗新生鼠缺氧缺血脑损伤作用的观察[J].第三军医大学学报,2006,28(20):2068-2070.
[16]黄丽.体外扩增的调节性T细胞静脉输注移植对脑梗死的保护作用[D].泰安:泰山医学院,2013.