基于PARP-1/AIF信号通路分析针刺对急性缺血性卒中大鼠脑血流及行为学恢复的影响
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国家自然科学基金资助项目(81603688);河北省卫建委医学科学研究基金项目(181218-4);河北省卫生健康委医学科学研究项目(20201226)


The effects of acupuncture on cerebral blood flow and behavioral recovery in acute ischemic stroke rats analyzed based on PARP-1/AIF signaling pathway
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    摘要:

    目的 基于PARP-1/AIF信号通路分析针刺对急性缺血性卒中大鼠脑血流及行为学恢复的影响。方法 将40只健康成年雄性SPF级大鼠分为Sham组、模型组、针刺组及针刺+PJ34组,每组10只,除Sham组外均建立动物模型,针刺组及针刺+PJ34组大鼠均进行针刺治疗,针刺+PJ34组以10 μmoL/g腹腔注射多聚腺苷二磷酸核糖转移酶-1(PARP-1)抑制剂PJ34,Sham组及模型组不进行针刺干预,均注射相同体积的0.9%生理盐水。11分制单盲评分评价建模后、干预第1、2、3天行为学评分;多普勒仪探头记录软脑膜微循环血流量;HE染色观察脑组织病理形态;TUNEL 法检测各组神经细胞凋亡率;免疫组化检测Bax、Bcl-2阳性表达;免疫印迹检测PARP-1、凋亡诱导因子(AIF)蛋白含量。结果 建模后,模型组、针刺组及针刺+PJ34组的行为学评分均升高,与Sham组比较差异有统计学意义(P<0.05),干预第1、2、3天时与模型组相比,针刺组行为学评分均降低(P<0.05),针刺+PJ34组上述时间行为学评分均降低,与针刺组比较差异有统计学意义(P<0.05);Sham组、模型组、针刺组及针刺+PJ34组的软脑膜微循环血流量分别为(756.35±102.33)、(233.06±45.17)、(416.40±63.55)、(560.80±72.04) AU,组间比较差异均有统计学意义(F=90.390,P<0.001);HE染色示Sham组大鼠脑组织及神经细胞排列整齐,模型组脑组织紊乱且神经细胞数目减少,针刺组及针刺+OJ34组神经细胞存活较多,排列整齐;Sham组、模型组、针刺组及针刺+PJ34组的神经细胞凋亡率分别为(3.30%±0.21%)、(30.04%±4.52%)、(16.30%±2.25%)、(11.69%±1.33%),组间比较差异均有统计学意义(F=90.390,P<0.001);与Sham组比较,模型组大鼠Bax阳性细胞数降低,Bcl-2阳性细胞数、PARP-1、AIF蛋白升高(均P<0.05),与模型组相比,针刺组及针刺+PJ34组的Bax阳性细胞数及Bcl-2阳性细胞数、PARP-1、AIF蛋白升高(均P<0.05),针刺组及针刺+PJ34组上述指标比较差异均有统计学意义(P<0.05)。结论 急性缺血性卒中大鼠采用针刺干预后能够改善行为学,增加脑血流灌溉的同时可通过调节细胞凋亡因子而减少神经细胞凋亡,研究机制可能与抑制PARP-1、AIF活性相关

    Abstract:

    Objective To analyze the effects of acupuncture on cerebral blood flow and behavioral recovery in rats with acute ischemic stroke based on PARP-1/AIF signaling pathway. Methods The rats were divided into SHAM group, model group, acupuncture group and acupuncture+PJ34 group, with 10 rats in each group. Animal models were established except sham group. The rats in acupuncture group and acupuncture +PJ34 group were treated with acupuncture, and the parP-1 inhibitor PJ34 was intraperitoneally injected with 10μmoL/g. Sham group and model group were injected 0.9% normal saline with the same volume without acupuncture intervention. Behavioral scores after modeling, intervention 1d, intervention 2D and intervention 3D were evaluated by 11-point single-blind score. The pia microcirculation blood flow was recorded by doppler probe. HE staining was used to observe the pathological morphology of brain tissue. TUNEL assay was used to detect the apoptosis rate of nerve cells in each group. Immunohistochemistry detected positive expression of Bax and Bcl-2. Western blot was used to detect the protein content of PARP-1 and AIF.Results After modeling, the behavioral scores of the model group, the acupuncture group and the acupuncture +PJ34 group increased, which was different from that of the SHAM group (P<0.05)Compared with the model group, the behavioral scores of the acupuncture group decreased on the first day, the second day and the third day of intervention (P<0.05)The behavioral scores of the acupuncture +PJ34 group decreased at the above time. There was a difference between acupuncture group and acupuncture group (P<0.05)The pia meningiae microcirculation blood flow of Sham group, model group, acupuncture group and acupuncture +PJ34 group were (756.35±102.33) AU, (233.06±45.17) AU, (416.40±63.55) AU and (560.80±72.04) AU, respectively, with differences (F=90.390, P<0.001). The brain tissue and nerve cells of sham group were arranged neatly, the brain tissue of model group was disordered and the number of nerve cells was reduced, the nerve cells of acupuncture group and acupuncture +OJ34 group were more survived and arranged neatly. The apoptosis rates of Sham group, model group, acupuncture group and acupuncture +PJ34 group were (3.30±0.21) %, (30.04±4.52) %, (16.30±2.25) % and (11.69±1.33) %, respectively (F=90.390, P<0.001)Compared with sham group, the number of Bax positive cells in model group was decreased, while the number of Bcl-2 positive cells, PARP-1 and AIF protein were increased (P<0.05). Compared with model group, the number of Bax positive cells, PARP-1 and AIF protein in acupuncture group and acupuncture +PJ34 group was decreased, while the bcl-2 positive, PARP-1 and AIF protein were increased (P<0.05)There were differences in above indexes between acupuncture group and acupuncture +PJ34 group (P<0.05)Conclusion Acupuncture can improve behavior, increase cerebral blood flow irrigation and reduce neuronal apoptosis by regulating apoptosis factors in rats with acute ischemic stroke. The mechanism may be related to inhibiting the activities of PARP-1 and AIF

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  • 在线发布日期: 2024-06-18
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