The Effect of Probiotics on the Expression of Aβ and the Protective Effect of Neurons in Rats with Cerebral Ischemia-Reperfusion Injury
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摘要:
目的 探讨益生菌干预对脑缺血再灌注损伤大鼠脑组织Aβ表达的影响及神经元的保护作用。 方法 实验动物为雄性Sprague-Dawley大鼠，随机分为4组，各组n = 12：假手术组、模型组、益生菌组、依达拉奉组。假手术组仅分离颈总动脉，其余各组使用线拴法建立脑缺血再灌注模型。脑损伤程度使用神经行为学评分评估；大鼠脑组织梗死面积使用TTC染色观察；大鼠大脑海马CA1区与皮层区神经元病理形态学改变特征使用HE染色观察；Aβ蛋白的表达使用免疫组化实验检测。 结果 模型组相较假手术组，神经行为学评分升高（P < 0.05），脑梗死面积增加（P < 0.05），海马CA1区与皮层区神经元出现损伤，Aβ蛋白表达显著上调（P < 0.001，P < 0.05）；益生菌组、依达拉奉组与模型组相比，神经行为学评分降低（P < 0.05），脑梗死面积减小（P < 0.05），海马CA1区与皮层区神经元损伤减轻，Aβ蛋白表达水平在两组中均显著下调（P < 0.001，P < 0.05）；依达拉奉组改善更为明显。 结论 益生菌干预可下调Aβ蛋白的表达，减轻脑缺血再灌注大鼠脑组织损伤、发挥神经元保护作用。 Abstract:Objective To explore the effect of probiotics interventionon of the expression of Aβ in rats with cerebral ischemia-reperfusion and the protective effect of neurons. Methods Male SD rats were selected as the experimental animals and randomly divided into Sham Group, Model Group, Probiotics Group and Edaravone Group.The Sham Group only separated the common carotid artery, while the other groups established a cerebral ischemia-reperfusion model using the suture method.The degree of brain injury was evaluated using neurobehavioral scoring; The infarct area of rat brain tissue was observed using TTC staining; HE staining was used to observe the pathological and morphological changes of neurons in the CA1 and cortical regions of the hippocampus in rats; The expression of Aβ proteins was detected using immunohistochemistry experiments. Results Compared with the Sham Group, the Model Group showed an increase in neurobehavioral scores （P < 0.05）, an increase in cerebral infarction focus （P < 0.05）, neuronal damage in the hippocampal CA1 and cortical regions, and a significant upregulation of Aβ protein expression （P < 0.001, P < 0.05）; Compared with the Model Group, the Probiotics Group and Edaravone Group showed a decrease in neurobehavioral scores （P < 0.05） and a reduction in cerebral infarction focus （P < 0.05）. The Probiotic Group and the Edaravone Group showed less neuronal damage in the hippocampal CA1 and cortical regions , and the Aβ protein expression levels were significantly down regulated in both groups （P < 0.001, P < 0.05）. The improvement was more significant in Edaravone Group. Conclusion Probiotics intervention can downregulate the expression of Aβ protein, alleviate the brain tissue damage and exert neuroprotective effects in rats with cerebral ischemia-reperfusion. -
Key words:
- Probiotics /
- Cerebral ischemia-reperfusion injury /
- β amyloid /
- Edaravone
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图 1 各组大鼠脑组织梗死灶
A：假手术组；B：模型组；C：益生菌组；D：依达拉奉组。
Figure 1. Cerebral infarction focus of rats in each group
图 2 各组大鼠大脑海马CA1区与皮层区神经元病理形态学改变（×400）
A～D：大鼠海马CA1区神经元病理形态学改变；E～H：大鼠皮层区神经元病理形态学改变。
Figure 2. Pathological and morphological changes of neurons in the hippocampus CA1 and cortical regions of rats in each group （×400）
图 3 各组大鼠大脑海马CA1区与皮层区Aβ蛋白的表达
A～D：大鼠海马CA1区神经元Aβ蛋白表达情况；E～H：大鼠皮层区神经元Aβ蛋白表达情况。
Figure 3. Expression of Aβ protein in the hippocampal CA1 and cortical regions of rats in each group
图 4 各组大鼠大脑海马CA1区与皮层区Aβ蛋白的表达比较
A：大鼠大脑海马CA1区Aβ蛋白的表达统计图；B：大鼠大脑皮层区Aβ蛋白的表达统计图与模型组比较，**P < 0.05，****P < 0.001。
Figure 4. Comparison of protein expression in the hippocampal CA1 and cortical regions of rats in each groups
表 1 Zea Longa 5 级评分法评分标准
Table 1. Zea longa grade 5 scoring method scoring standard
评分 表现 0 无神经损害症状 1 梗死侧对侧前肢和前爪不能完全伸展 2 不能正常行走，行走时向梗死侧旋转 3 行走时向梗死对侧倾倒 4 无法行走和意识障碍 
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表 2 各组大鼠神经行为学评分[（$ \bar x \pm s $），分]
Table 2. Neurobehavioral scores of rats in each group[ （$ \bar x \pm s $），points]
分组 n 神经行为学评分 假手术组 12 0±0* 模型组 12 3.508±0.412 益生菌组 12 2.53±0.41* 依达拉奉组 12 2.274±0.196* 与模型组比较，*P < 0.05。
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表 3 各组大鼠脑组织梗死灶体积的比较[（$ \bar x \pm s$）%]
Table 3. Comparison of cerebral infarction focus volume of rats in each group [（$ \bar x \pm s$）%]
分组 n 脑梗死体积百分比 假手术组 3 0±0* 模型组 3 21.558±2.238 益生菌组 3 9.482±1.428* 依达拉奉组 3 6.988±1.322* 与模型组比较，*P < 0.05。
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[1] Savitz S I,Baron J C,Yenari M A,et al. Reconsidering neuroprotection in the reperfusion era[J]. Stroke,2017,48(12):3413-3419. doi: 10.1161/STROKEAHA.117.017283 [2] Pawar A,Pardasani K R. Mechanistic insights of neuronal calcium and ip3 signaling system regulating atp release during ischemia in progression of Alzheimer's disease[J]. Eur Biophys J,2023,52(3):153-173. doi: 10.1007/s00249-023-01660-1 [3] Ren H,Ma L,Gong X,et al. Edaravone exerts brain protective function by reducing the expression of aqp4,app and aβ proteins[J]. Open Life Sci,2019,14(1):651-658. doi: 10.1515/biol-2019-0074 [4] Pluta R,Ułamek-Kozioł M,Januszewski S,et al. Participation of amyloid and tau protein in neuronal death and neurodegeneration after brain ischemia[J]. Int J Mol Sci,2020,21(13):4599. doi: 10.3390/ijms21134599 [5] Ułamek-Kozioł M,Czuczwar S J,Januszewski S,et al. Proteomic and genomic changes in tau protein,which are associated with Alzheimer's disease after ischemia-reperfusion brain injury[J]. Int J Mol Sci,2020,21(3):892. doi: 10.3390/ijms21030892 [6] Nguyen T-V V,Hayes M,Zbesko J C,et al. Alzheimer's associated amyloid and tau deposition co-localizes with a homeostatic myelin repair pathway in two mouse models of post-stroke mixed dementia[J]. Acta Neuropathol Commun,2018,6(1):100. doi: 10.1186/s40478-018-0603-4 [7] Wang J,Zhang H,He J,et al. The role of the gut microbiota in the development of ischemic stroke[J]. Front Immunol,2022,28(13):845243. [8] Benakis C,Poon C,Lane D,et al. Distinct commensal bacterial signature in the gut is associated with acute and long-term protection from ischemic stroke[J]. Stroke,2020,51(6):1844-1854. doi: 10.1161/STROKEAHA.120.029262 [9] Li H,Sun J,Du J,et al. Clostridium butyricum exerts a neuroprotective effect in a mouse model of traumatic brain injury via the gut-brain axis[J]. Neurogastroenterol Motil,2018,30(5):e13260. doi: 10.1111/nmo.13260 [10] 冯云,亢君君,方宗平,等. 肠道菌群移植通过降低il-17水平减轻老年小鼠脑缺血再灌注损伤[J]. 细胞与分子免疫学杂志,2019,35(1):52-57. [11] Wang H,Ren S,Lv H,et al. Gut microbiota from mice with cerebral ischemia-reperfusion injury affects the brain in healthy mice[J]. Aging (Albany NY),2021,13(7):10058-10074. doi: 10.18632/aging.202763 [12] Wu H,Chiou J. Potential benefits of probiotics and prebiotics for coronary heart disease and stroke[J]. Nutrients,2021,13(8):2878. doi: 10.3390/nu13082878 [13] Pan F,Zhang L,Li M,et al. Predominant gut lactobacillus murinus strain mediates anti-inflammaging effects in calorie-restricted mice[J]. Microbiome,2018,6(1):54. doi: 10.1186/s40168-018-0440-5 [14] Akhoundzadeh K,Vakili A,Shadnoush M,et al. Effects of the oral ingestion of probiotics on brain damage in a transient model of focal cerebral ischemia in mice[J]. Iran J Med Sci,2018,43(1):32-40. [15] 刘兆兰,刘宝全,李兰花,等. 动态随机分组方法介绍及应用[J]. 中西医结合学报,2011,9(3):246-251. [16] 梁国晶,任海燕,张钰鸽,等. 红茶菌在脑缺血再灌注损伤大鼠模型中的作用研究[J]. 现代生物医学进展,2023,23(3):428-432. [17] 卢小叶,吕倩忆,李棋龙,等. Zea-longa评分与改良Garcia评分应用于针刺治疗CIRI大鼠神经功能缺损评估的研究[J]. 湖南中医药大学学报,2021,41(9):1356-1360. [18] Li Y,Zhang J. Animal models of stroke[J]. Animal Model Exp Med,2021,4(3):204-219. doi: 10.1002/ame2.12179 [19] Maida C D,Norrito R L,Daidone M,et al. Neuroinflammatory mechanisms in ischemic stroke: Focus on cardioembolic stroke,background,and therapeutic approaches[J]. Int J Mol Sci,2020,21(18):6454. doi: 10.3390/ijms21186454 [20] Dai S J,Zhang J Y,Bao Y T,et al. Intracerebroventricular injection of aβ1-42 combined with two-vessel occlusion accelerate Alzheimer's disease development in rats[J]. Pathol Res Pract,2018,214(10):1583-1595. doi: 10.1016/j.prp.2018.07.020 [21] Pluta R,Miziak B,Czuczwar S J. Post-ischemic permeability of the blood-brain barrier to amyloid and platelets as a factor in the maturation of alzheimer's disease-type brain neurodegeneration[J]. Int J Mol Sci,2023,24(13):10739. doi: 10.3390/ijms241310739 [22] Loh J S, Mak W Q, Tan L K S, et al. Microbiota-gut-brain axis and its therapeutic applications in neurodegenerative diseases[J]. Signal Transduct Target Ther,2024,9(1):37. Published 2024 Feb 16. [23] Abdelhamid M,Zhou C,Ohno K,et al. Probiotic bifidobacterium breve prevents memory impairment through the reduction of both amyloid-β production and microglia activation in app knock-in mouse[J]. J Alzheimers Dis,2022,85(4):1555-1571. doi: 10.3233/JAD-215025 [24] 毛丙永,崔树茂,潘明罗,等. 复合益生菌制剂对人体肠道菌群组成的调节作用[J]. 现代食品科技,2021,37(11):8-13,113. [25] 杨梅芳,程萍,陈治任,等. 肠道菌群代谢产物TMAO激活HMGB1/NLRP3炎症通路促进小鼠脑缺血半暗带损伤的机制研究[J]. 中风与神经疾病杂志,2023,40(12):1096-1100. [26] Gilbert K,Arseneault-Br é ard J,Flores Monaco F,et al. Attenuation of post-myocardial infarction depression in rats by n-3 fatty acids or probiotics starting after the onset of reperfusion[J]. Br J Nutr,2013,109(1):50-56. doi: 10.1017/S0007114512003807 [27] Rezaeiasl Z,Salami M,Sepehri G. The effects of probiotic lactobacillus and bifidobacterium strains on memory and learning behavior,long-term potentiation (ltp),and some biochemical parameters in β-amyloid-induced rat's model of alzheimer's disease[J]. Prev Nutr Food Sci,2019,24(3):265-273. doi: 10.3746/pnf.2019.24.3.265 [28] Kim M S,Kim Y,Choi H,et al. Transfer of a healthy microbiota reduces amyloid and tau pathology in an Alzheimer's disease animal model[J]. Gut,2020,69(2):283-294. doi: 10.1136/gutjnl-2018-317431 [29] 任海燕,赵晖,王蕾,等. 依达拉奉对脑缺血再灌注后aβ及其前体表达干预[J]. 科技导报,2015,33(12):77-82. [30] Ye T,Yuan S,Kong Y,et al. Effect of probiotic fungi against cognitive impairment in mice via regulation of the fungal microbiota-gut-brain axis[J]. J Agric Food Chem,2022,70(29):9026-9038. doi: 10.1021/acs.jafc.2c03142 -
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