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| 不同穴位电针刺激联合hUC-MSCs对缺血性脑损伤大鼠脑屏障、腺苷A2A受体及GSK-3β/β-catenin表达的影响 |
| Effects of electroacupuncture at different points combined with huc-Mscs on the expression of brain barrier, adenosine A2A receptor and GSK-3β/β-catenin in rats with ischemic brain injury |
| 投稿时间：2022-05-04 录用日期：2022-05-26 |
| DOI： |
| 中文关键词: 不同穴位电针刺激、hUC-MSCs、缺血性脑损伤大鼠、脑屏障、腺苷A2A受体、GSK-3β/β-catenin |
| 英文关键词: Electroacupuncture stimulation at different acupoints, hec-Mscs, brain barrier, adenosine A2A receptor, GSK-3β/β-catenin, ischemic brain injury rats |
| 基金项目:国家自然科学(编号：81704148) |
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| 中文摘要: |
| 目的：研究不同穴位电针刺激联合hUC-MSCs对缺血性脑损伤大鼠脑屏障、腺苷A2A受体及GSK-3β/β-catenin表达的影响。方法：健康SD大鼠40只购自北京博尔西科技有限公司,随机分为健康组、模型组(脑损伤大鼠)、移植组(模型+hUC-MSCs移植)、联合组(模型+hUC-MSCs移植+电针刺激百会、风府、肾俞穴),每组10只。HE染色观察组织形态学。TUNEL染色检测脑细胞凋亡。免疫荧光检测A2A受体。免疫组化检测β-catenin、GSK-3β表达。免疫印迹检测occludin 、 ZO-1 蛋白表达。结果：BrdU 标记 hUC-MSCs,荧光显微镜下可见红色荧光,证实该细胞为人脐带间充质干细胞。HE 染色结果显示,健康组海马 CA1 区组织结构正常,锥体细胞排列整齐,而模型组海马组织结构明显异常,锥体细胞排列紊乱,细胞间隙增宽,神经元变性明显,部分退变的神经元呈现凋亡特征,细胞皱缩成圆形或卵圆形、核染色质固缩、边集或碎裂。而移植组与联合组神经细胞排列略紊乱,仍可见细胞皱缩和胞核固缩浓染现象,但程度较模型组明显减轻,且以联合组效果更加。与健康组相比,模型组脑细胞凋亡率、GSK-3β升高,occludin 、 ZO-1、β-catenin降低(P<0.05)。与模型组相比,移植组细胞凋亡率、GSK-3β降低,occludin 、 ZO-1、β-catenin升高(P<0.05)。与移植组相比,联合组细胞凋亡率、GSK-3β降低,occludin 、 ZO-1、β-catenin升高(P<0.05)。结论：不同穴位电针刺激联合hUC-MSCs可改善缺血性脑损伤大鼠脑屏障； 抑制苷A2A受体、GSK-3β水平,提高β-catenin表达,抑制脑组织细胞凋亡,起到脑保护作用。 |
| 英文摘要: |
| Objective: To investigate the effects of electroacupuncture at different acupoints combined with hec-Mscs on the expression of brain barrier, adenosine A2A receptor and GSK-3β/β-catenin in rats with ischemic brain injury. Methods: forty healthy SD rats were purchased from Beijing Boxi Science and Technology Co., LTD., and were randomly divided into healthy group, model group (brain injury rats), transplantation group (model + huc-Mscs transplantation), combined group (model + huc-Mscs transplantation + electroacupuncture stimulation at Baihui, Fengfu and Shenshu), with 10 rats in each group. Histomorphology was observed by HE staining. TUNEL staining was used to detect apoptosis of brain cells. A2A receptor was detected by immunofluorescence. The expression of β-catenin and GSK-3β was detected by immunohistochemistry. The expression of Occludin and ZO-1 protein was detected by western blot. Results: THE BrdU labeled HUC-Mscs showed red fluorescence under fluorescence microscope, confirming that the SCS were human umbilical cord mesenchymal stem cells. HE staining, according to the results of hippocampal CA1 area structure normal healthy group, neat cone cells, and hippocampal structure model group was obviously abnormal, disordered arrangement of pyramidal cells and intercellular broadening, neuron degeneration, part of the degeneration of neurons apoptosis characteristics, cells shrivel round or oval, blue-stained nuclear chromatin pyknosis, side set or fracture. However, the arrangement of nerve cells in the transplantation group and the combined group was slightly disordered, and the phenomenon of cell shrinkage and nucleus hyperchromatism were still observed, but the degree was significantly less than that in the model group, and the effect was more obvious in the combined group. Compared with healthy group, the apoptosis rate and GSK-3β of brain cells in model group increased, while occludin, ZO-1 and β-catenin decreased (P<0.05). Compared with model group, the apoptosis rate and GSK-3β in transplantation group decreased, while occludin, ZO-1 and β-catenin increased (P<0.05). Compared with transplantation group, the apoptosis rate and GSK-3β in combined group decreased, while occludin, ZO-1 and β-catenin increased (P<0.05). Conclusion: Electroacupuncture at different points combined with HUC-Mscs can improve the brain barrier in rats with ischemic brain injury. Inhibit glycoside A2A receptor and GSK-3β, increase the expression of β-catenin, inhibit the apoptosis of brain tissue cells, and play a protective effect on brain. |
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