ADSCs通过PI3K/Akt和NF-κB途径调控神经元存活及炎症反应  

ADSCs regulate neuronal survival and inflammatory response through PI3K/Akt and NF-kB pathways

作　　者：李伟宽 周治来 谢欢[3] 张辉 Li Weikuan;Zhou Zhilai;Xie Huan;Zhang Hui(Second Clinical College of Southern Medical University,Guangzhou 510220,China)

机构地区：[1]南方医科大学第二临床医学院,广州510220 [2]广东省第二人民医院脊柱骨科,广州510220 [3]广州暨南大学第一临床学院

出　　处：《脑与神经疾病杂志》2025年第1期1-7,共7页Journal of Brain and Nervous Diseases

基　　金：广东省自然科学基金(2016A030313627);广州市科技计划项目(202002030004);广州市科技计划项目(202201010803)。

摘　　要：目的探究体外脂肪间充质干细胞(ADSCs)对神经元的保护作用及机制。方法培养并鉴定原代ADSCs,并建立谷氨酸诱导的HT22神经元细胞系损伤模型。细胞免疫荧光检测损伤模型中HT22细胞分支数目及分支长度的变化。构建ADSCs与HT22共培养,检测ADSCs对HT22细胞凋亡的调节作用。采用ELISA研究ADSCs对HT22损伤细胞中炎症因子(TNF-α、IL-1β、IL-6、IL-10)的影响。通过Western blot检测PI3K/Akt及NF-κB的磷酸化变化,探讨ADSC对损伤HT22的机制。结果细胞免疫荧光验证酶解法培养的ADSCs高表达CD44。相较于对照组,细胞毒性试验表明随着谷氨酸增高,HT22细胞存活率下降,且20mM为半数抑制浓度,该浓度下HT22支数目及分支长度显著减少。相较于谷氨酸组,ADSCs共培养有效降低了谷氨酸诱导的HT22细胞系突起的分细胞凋亡,减少了Cleaved caspase3的表达。ELISA结果显示ADSCs共培养显著降低损伤HT22细胞中促炎因子的表达,同时提高抗炎因子IL-10的水平。ADSCs通过促进PI3K/Akt的磷酸化和抑制NF-κB的磷酸化,调节神经元存活及炎症反应。结论ADSCs具有显著的神经保护作用,通过调节关键信号途径促进受损神经元的存活和功能恢复。这为脊髓损伤的治疗提供了新的研究方向和希望。Objective This study aimed to investigate the neuroprotective effects and mechanisms of adiposederived mesenchymal stem cells(ADSCs)on neurons in vitro.Methods Primary ADSCs were cultured and identified,and a glutamate-induced HT22 neuronal cell injury model was established.Cell immunofluorescence was used to detect changes in the number and length of branches in the injured HT22 model.Co-cultures of ADSCs and HT22were established to examine the regulatory effects of ADSCs on HT22 cell apoptosis.ELISA was employed to study the impact of ADSCs on inflammatory factors(TNF-α,IL-1β,IL-6,IL-10)in injured HT22 cells.Western blot analysis was performed to investigate changes in PI3K/Akt and NF-κB phosphorylation,exploring the mechanisms of ADSCs action on injured HT22.Results Cell immunofluorescence confirmed that enzymatic culture of ADSCs expressed high levels of CD44.Compared to the control group,cytotoxicity assays indicated a gradual decrease in HT22 cell survival with increasing glutamate concentration,reaching 20mM as the half-maximal inhibitory concentration.At this concentration,HT22 branch number and length significantly decreased.ADSCs effectively reduced glutamate-induced HT22 cell apoptosis and decreased cleaved caspase3 expression compared to the Glu group.ELISA results showed that ADSCs significantly decreased the expression of pro-inflammatory factors in injured HT22 cells while increasing the level of the anti-inflammatory factor IL-10.ADSCs regulated neuronal survival and inflammatory responses through promoting PI3K/Akt phosphorylation and inhibiting NF-κB phosphorylation.Conclusion ADSCs demonstrate significant neuroprotective effects,promoting the survival and functional recovery of damaged neurons by modulating key signaling pathways.This discovery provides new research directions and hope for the treatment of spinal cord injuries(SCI).

关 键 词：脊髓损伤 脂肪间充质干细胞 HT22 谷氨酸 

分 类 号：R744[医药卫生—神经病学与精神病学]