针刺对缺氧缺血性脑损伤新生大鼠的神经保护作用及对小胶质细胞和炎症反应的影响

Neuroprotective effects of acupuncture on neonatal rats with hypoxic-ischemic brain damage and their effects on microglia and inflammatory response

目的:探讨针刺对缺氧缺血性脑损伤新生大鼠的神经保护作用及对小胶质细胞和炎症反应的影响。方法:将150只大鼠根据随机数字法分为假手术组、模型组(建立缺氧缺血性脑损伤模型)和针刺组(建立缺氧缺血性脑损伤模型+针刺治疗),每组50只。采用2,3,5-三苯基氯化四氮唑(TTC)染色法测定脑梗死情况,采用免疫组化法测定脑组织小胶质细胞标记物离子钙接头分子(Iba-1)表达,采用逆转录-聚合酶链反应(RT-PCR)测定脑组织IL-6、IL-1β、TNF-αmRNA表达水平,采用ELISA法测定脑组织IL-6、IL-1β、TNF-α含量水平,采用Morris水迷宫测定大鼠空间学习记忆能力。结果:与假手术组比较,模型组和针刺组大鼠逃避潜伏期延长(P<0.05),穿台指数降低(P<0.05);与模型组比较,针刺组大鼠逃避潜伏期缩短(P<0.05),穿台指数增加(P<0.05)。假手术组大鼠无梗死现象,针刺组脑梗死面积明显低于模型组(P<0.05)。与假手术组比较,模型组和针刺组大鼠脑组织Iba-1阳性细胞增多(P<0.05),与模型组比较,针刺组大鼠脑组织Iba-1阳性细胞减少(P<0.05)。与假手术组比较,模型组和针刺组大鼠脑组织IL-6、IL-1β、TNF-α含量水平及mRNA相对表达量升高(P<0.05);与模型组比较,针刺组大鼠脑组织IL-6、IL-1β、TNF-α含量水平及mRNA相对表达量降低(P<0.05)。结论:针刺对缺氧缺血性脑损伤新生大鼠有神经保护作用,其机制可能与抑制小胶质细胞活化和降低脑组织炎症反应有关。

Objective:To investigate the neuroprotective effects of acupuncture on neonatal rats with hypoxic-ischemic brain damage and their effects on microglia and inflammatory response.Methods:150 rats were divided into sham operation group,model group(established hypoxic ischemic brain injury model) and acupuncture group(established hypoxic ischemic brain injury model +acupuncture treatment) accordedto random number method,50 cases in each group.Cerebral infarction was measured by 2,3,5-triphenyltetrazolium chloride(TTC) staining.Immunohistochemistry was used to determine the expression of microglia marker ion calcium linker molecule(Iba-1) in brain tissue.The levels of IL-6,IL-1β and TNF-α mRNA in brain tissue were determined by reverse transcription-polymerase chain reaction(RT-PCR).The levels of interleukin-6(IL-6),interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) in brain tissue were determined by enzyme-linked immunosorbent assay(ELISA).The spatial learning and memory ability of rats were deter mined by Morris water maze.Results:Compared with the sham operation group,the escape latency of the model group and the acupuncture group were prolonged(P<0.05),and the penetration index were decreased(P<0.05).Compared with the model group,the escape latency of the acupuncture group was shortened(P<0.05),and the penetration index was increased(P<0.05).There was no infarction in the sham operation group.The infarct size in the acupuncture group was significantly lower than that in the model group(P<0.05).Compared with the sham operation group,the Iba-1 positive cells in the brain tissue of the model group and the acupuncture group were increased(P<0.05).Compared with the model group,the Iba-1 positive cells in the brain tissue of rats in the acupuncture group was decreased(P<0.05).Compared with the sham operation group,the levels and the mRNA expression of IL-6,IL-1β,TNF-α in the brain tissue of the model group and the acupuncture group were increased(P<0.05).Compared with the model group,the levels and the mRNA expression of in the acupuncture group were decreased(P<0.05).Conclusion:Acupuncture has neuroprotective effects on neonatal rats with hypoxic-ischemic brain damage,and its mechanism may be related to inhibition of microglia activation and reduction of brain tissue inflammation.