中国组织工程研究

中国组织工程研究 ›› 2024, Vol. 28 ›› Issue (31): 5036-5041.doi: 10.12307/2024.709

• 干细胞移植 stem cell transplantation • 上一篇    下一篇

颅内移植人骨髓间充质干细胞减轻大鼠脑缺血再灌注损伤

宋文学1，廖益东1，明  江1，何龙才1，陈光唐1，陈  晨1，王梓力1，熊明松1，崔君拴2，徐卡娅2，3   

  1. 1贵州医科大学，贵州省贵阳市   550004；贵州医科大学附属医院，2神经外科，3高压氧科，贵州省贵阳市   550004
  • 收稿日期:2023-07-24 接受日期:2023-10-12 出版日期:2024-11-08 发布日期:2024-01-22
  • 通讯作者: 徐卡娅，博士，主任医师，贵州医科大学附属医院，神经外科，高压氧科，贵州省贵阳市 550004
  • 作者简介:宋文学，男，1995年生，河南省信阳市人，汉族，贵州医科大学在读硕士，主要从事人骨髓间充质干细胞移植治疗缺血性脑卒中的研究。
  • 基金资助:
    国家自然科学基金项目(81901173，82060231)，项目负责人：徐卡娅；贵州医科大学附属医院博士科研启动基金项目(gyfybsky-2021-6)，项目负责人：徐卡娅；2022年国家自然科学基金培育项目计划(gyfynsfc-2022-08)，项目负责人：徐卡娅；贵州省科技计划项目(ZK【2023】重点039)，项目负责人：徐卡娅

Intracranial transplantation of human bone marrow mesenchymal stem cells alleviates rat brain ischemia-reperfusion injury

Song Wenxue1, Liao Yidong1, Ming Jiang1, He Longcai1, Chen Guangtang1, Chen Chen1, Wang Zili1, Xiong Mingsong1, Cui Junshuan2, Xu Kaya2, 3   

  1. 1Guizhou Medical University, Guiyang 550004, Guizhou Province, China; 2Department of Neurosurgery, 3Department of Hyperbaric Oxygen, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China
  • Received:2023-07-24 Accepted:2023-10-12 Online:2024-11-08 Published:2024-01-22
  • Contact: Xu Kaya, MD, Chief physician, Department of Neurosurgery, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China; Department of Hyperbaric Oxygen, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China
  • About author:Song Wenxue, Master candidate, Guizhou Medical University, Guiyang 550004, Guizhou Province, China
  • Supported by:
    National Natural Science Foundation of China, Nos. 81901173 and 82060231 (to XKY); Doctoral Research Start-up Fund Project of Affiliated Hospital of Guizhou Medical University, No. gyfybsky-2021-6 (to XKY); Cultivation Project Plan of National Natural Science Foundation of China in 2022, No. gyfynsfc-2022-08 (to XKY); Science and Technology Plan Project of Guizhou Province, No. ZK [2023] Key 039 (to XKY)

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摘要：


文题释义：

脑缺血再灌注损伤：指脑组织在一段时间的缺血(血液供应不足)后，恢复血液供应时会出现一系列的病理生理过程，如氧化应激、能量代谢紊乱、细胞损伤、炎症反应等，这些过程可能对脑组织造成直接或间接的损害。
氧化应激：是指细胞内氧化还原平衡失调，导致氧化物(例如活性氧化物和自由基)产生过多，超过细胞的抗氧化能力所能消除的状态。这种失衡状态可以引起细胞膜的脂质过氧化、蛋白质氧化、核酸损伤等，损害细胞结构和功能。


背景：研究发现激活Nrf2/HO-1通路可减轻脑缺血再灌注损伤引起的氧化应激，但人骨髓间充质干细胞(human bone marrow 

mesenchymal stem cells，hBMSC)是否可激活Nrf2/HO-1通路减轻脑缺血再灌注损伤目前尚缺乏相关研究。
目的：探讨颅内移植hBMSC是否通过激活Nrf2/HO-1途径减轻脑缺血再灌注模型大鼠的氧化应激损伤。
方法：40只雄性SPF级SD大鼠随机分为假手术组、模型组、hBMSC移植组、hBMSC+溶剂组和hBMSC+Nrf2抑制剂组，每组8只。模型组、hBMSC移植组通过线栓法制备大脑中动脉栓塞模型，1 h后取出线栓，将30 μL PBS或培养至5代以后的hBMSC注入大鼠右侧皮质及纹状体内；hBMSC+Nrf2抑制剂组和hBMSC+溶剂组在造模前24 h于左侧脑室分别注射Nrf2抑制剂Brusatol或其溶剂二甲基亚砜，然后制备大脑中动脉栓塞模型，注射hBMSC。移植后3 d进行相关指标检测。

结果与结论：①脑CT和TTC染色显示脑梗死灶面积和体积一致：模型组> hBMSC+Nrf2抑制剂组> hBMSC+溶剂组> hBMSC移植组>假手术组；②苏木精-伊红染色和尼氏染色显示，假手术组缺血侧脑组织形态完好和神经元正常；模型组神经元排列结构紊乱和神经元损害严重；与模型组相比，hBMSC移植组、hBMSC+溶剂组病理形态和神经元损伤有较大的改善；与hBMSC+溶剂组相比，hBMSC+Nrf2抑制剂组病理形态和神经元损伤更严重；③Western blot和氧化应激指标检测结果显示，与假手术组相比，模型组Nrf2和HO-1蛋白表达降低(均P < 0.05)，丙二醛升高而超氧化物歧化酶降低(均P < 0.05)；与模型组相比，hBMSC移植组和hBMSC+溶剂组Nrf2、HO-1蛋白表达升高(均P < 0.05)，丙二醛降低而超氧化物歧化酶升高(均P < 0.05)；与hBMSC+溶剂组相比，hBMSC+Nrf2抑制剂组Nrf2和HO-1蛋白表达降低(均P < 0.05)，丙二醛升高而超氧化物歧化酶降低(均P < 0.05)；④上述结果表明，hBMSC可能通过激活Nrf2/HO-1途径减轻脑缺血再灌注氧化应激损伤。

https://orcid.org/0009-0008-1478-3444 (宋文学)

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 人骨髓间充质干细胞, 脑缺血再灌注损伤, 氧化应激, 核因子E2相关因子2, 血红素加氧化酶1

Abstract: BACKGROUND: Studies have found that activation of nuclear factor-erythroid 2-related factor 2/heme oxidase-1 (Nrf2/HO-1) pathway can alleviate oxidative stress caused by cerebral ischemia-reperfusion injury, but whether human bone marrow mesenchymal stem cells (hBMSC) can activate Nrf2/HO-1 pathway to alleviate cerebral ischemia-reperfusion injury is still lacking relevant studies.
OBJECTIVE: To investigate whether intracranial transplantation of hBMSC alleviates oxidative stress injury in cerebral ischemia-reperfusion animal models by activating Nrf2/HO-1 pathway.
METHODS: Totally 40 male SPF SD rats were randomly divided into sham operation group, model group, hBMSC transplantation group, hBMSC+solvent group and hBMSC+Nrf2 inhibitor group. Each group consisted of eight animals. In the model group and the hBMSC transplantation group, middle cerebral artery occlusion model was prepared by thread embolization method. The thread embolization was removed 1 hour later, and 30 μL PBS or hBMSC cultured to at least passage 5 was injected into the right cortex and striatum of rats. In the hBMSC+Nrf2 inhibitor group and hBMSC+solvent group, the left ventricle was injected with Nrf2 inhibitor Brusatol and its solvent dimethyl sulfoxide respectively 24 hours before model establishment, then the middle cerebral artery occlusion model was prepared, and hBMSC was injected. Relevant indexes were detected 3 days after transplantation.
RESULTS AND CONCLUSION: (1) CT and TTC staining showed the same area and volume of cerebral infarction: model group > hBMSC+Nrf2 inhibitor group > hBMSC+solvent group > hBMSC transplantation group > sham operation group. (2) Hematoxylin-eosin staining and Nissl’s staining showed that the ischemic brain tissue was intact and the neurons were normal in the sham operation group. Compared with the model group, the pathological morphology and neuronal injury of the hBMSC transplantation group and the hBMSC+solvent group were significantly improved. Compared with the hBMSC+solvent group, the hBMSC+Nrf2 inhibitor group had more serious pathological morphology and neuronal damage. (3) Western blot assay and oxidative stress index detection results showed that compared with the sham operation group, Nrf2 and HO-1 proteins were decreased (all P < 0.05), malondialdehyde was increased and superoxide dismutase was decreased (all P < 0.05) in the model group. Compared with the model group, the expression levels of Nrf2 and HO-1 proteins were increased (all P < 0.05), malondialdehyde was decreased and superoxide dismutase was increased (all P < 0.05) in the hBMSC transplantation group and the hBMSC+solvent group. Compared with the hBMSC+solvent group, the expression levels of Nrf2 and HO-1 proteins were simultaneously decreased (all P < 0.05), and malondialdehyde was increased and superoxide dismutase was decreased (all P < 0.05) in the hBMSC+Nrf2 inhibitor group. (4) These results indicate that hBMSC can alleviate cerebral ischemia-reperfusion injury possibly by activating Nrf2/HO-1 pathway.

Key words: human bone marrow mesenchymal stem cell, cerebral ischemia-reperfusion injury, oxidative stress, nuclear factor-erythroid 2-related factor 2, heme oxidase-1

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