PDF(Pubmed)
Abstract：
Adipose-derived stem cells (ASCs) possess therapeutic potential for ischemic brain injury, and the chemokine CXCL12 has been shown to enhance their functional properties. However, the cumulative effects of ASCs when combined with various structures of CXCL12 on ischemic stroke and its underlying molecular mechanisms remain unclear. In this study, we genetically engineered mouse adipose-derived ASCs with CXCL12 variants and transplanted them to the infarct region in a mice transient middle cerebral artery occlusion (tMCAO) model of stroke. We subsequently compared the post-ischemic stroke efficacy of ASC-mCXCL12 with ASC-dCXCL12, ASC-wtCXCL12, and unmodified ASCs. Neurobehavior recovery was assessed using modified neurological severity scores, the hanging wire test, and the elevated body swing test. Changes at the tissue level were evaluated through cresyl violet and immunofluorescent staining, while molecular level alterations were examined via Western blot and real-time PCR. The results of the modified neurological severity score and cresyl violet staining indicated that both ASC-mCXCL12 and ASC-dCXCL12 treatment enhanced neurobehavioral recovery and mitigated brain atrophy at the third and fifth weeks post-tMCAO. Additionally, we observed that ASC-mCXCL12 and ASC-dCXCL12 promoted angiogenesis and neurogenesis, accompanied by an increased expression of bFGF and VEGF in the peri-infarct area of the brain. Notably, in the third week after tMCAO, the ASC-mCXCL12 exhibited superior outcomes compared to ASC-dCXCL12. However, when treated with the CXCR4 antagonist AMD3100, the beneficial effects of ASC-mCXCL12 were reversed. The AMD3100-treated group demonstrated worsened neurological function, aggravated edema volume, and brain atrophy. This outcome is likely attributed to the interaction of monomeric CXCL12 with CXCR4, which regulates the recruitment of bFGF and VEGF. This study introduces an innovative approach to enhance the therapeutic potential of ASCs in treating ischemic stroke by genetically engineering them with the monomeric structure of CXCL12.
摘要：
脂肪来源的干细胞（ASCs）具有治疗缺血性脑损伤的潜力，趋化因子CXCL12已被证明可以增强其功能特性。然而,ASCs与CXCL12多种结构联合对缺血性卒中的累积效应及其潜在分子机制尚不清楚.在这项研究中,我们将小鼠脂肪来源的ASCs与CXCL12变体进行基因工程改造,并将其移植到小鼠短暂性大脑中动脉闭塞(tMCAO)卒中模型的梗死区.我们随后比较了ASC-mCXCL12与ASC-dCXCL12、ASC-wtCXCL12和未修饰的ASCs的缺血性卒中后疗效。使用改良的神经严重程度评分评估神经行为恢复，吊线测试，和升高的身体摆动测试。通过甲酚紫和免疫荧光染色评估组织水平的变化，同时通过蛋白质印迹和实时PCR检查分子水平的改变。改良的神经学严重程度评分和甲酚紫染色的结果表明，在tMCAO后的第三和第五周，ASC-mCXCL12和ASC-dCXCL12治疗均增强了神经行为恢复并减轻了脑萎缩。此外,我们观察到ASC-mCXCL12和ASC-dCXCL12促进血管生成和神经发生，伴随着bFGF和VEGF在脑梗塞周围区域的表达增加。值得注意的是,在tMCAO之后的第三周，与ASC-dCXCL12相比,ASC-mCXCL12表现出更好的结局.然而,当用CXCR4拮抗剂AMD3100治疗时,ASC-mCXCL12的有益作用被逆转.AMD3100治疗组表现出神经功能恶化，加重水肿体积，和脑萎缩.该结果可能归因于单体CXCL12与CXCR4的相互作用,其调节bFGF和VEGF的募集。这项研究引入了一种创新的方法，通过用CXCL12的单体结构对ASCs进行基因工程改造来增强ASCs治疗缺血性中风的治疗潜力。
