目的: 研究睫状体营养因子(CNTF)对氧糖剥夺(OGD)模型小鼠海马神经元细胞(HT22)凋亡的影响。方法: 将更换为无糖培养基的HT22细胞置于低氧培养箱中培养6 h,转移至常氧培养箱复氧18 h。观察细胞形态变化,检测凋亡相关因子蛋白的表达,检测凋亡相关因子mRNA表达,转染质粒(sh-nc、sh-cntf、oe-cntf)及抑制剂(sh-akt)至HT22细胞后更换无糖培养基置于低氧培养箱中培养6 h,转移至常氧培养箱复氧18 h。检测各组HT22凋亡相关因子及AKT/STAT3蛋白表达。结果: 与正常组相比,OGD处理组细胞凋亡水平提高、抗凋亡能力降低,镜下观察OGD组细胞胞体空泡明显增多;与正常组相比,OGD组与sh-nc+OGD组细胞凋亡水平明显升高且两组之间凋亡水平无明显差异,sh-cntf组细胞凋亡水平进一步升高,但在过表达(oe-cntf)组中凋亡水平又明显下降,AKT敲低组(sh-akt)与AKT、CNTF共同敲低组(sh-akt+cntf)相比于正常组细胞凋亡水平明显上升并且sh-akt+cntf组中凋亡水平上升的更为明显,在敲低AKT与过表达CNTF(sh-akt+oe-cntf)后凋亡得到了抑制。结论: CNTF通过激活AKT/STAT3信号通路促进STAT3中705号磷酸位点的磷酸化从而抑制OGD后神经元细胞的凋亡从而发挥保护作用。
林家静
,
青格尔
,
焦定一
,
谢伟
,
谢雅彬
,
许文强
,
石蕊
,
贾小娥
,
姜树原
,
刘晓蕾
,
鲍牧兰
,
巴德仁贵
. CNTF调控AKT/STAT3信号通路影响氧糖剥夺模型神经元细胞的凋亡水平*[J]. 包头医学院学报, 2025
, 41(12)
: 10
-15
.
DOI: 10.16833/j.cnki.jbmc.2025.12.003
Objective: To study the effect of ciliary neurotrophic factor (CNTF) on the apoptosis of hippocampal neurons (HT22) in oxygen-glucose deprivation (OGD) model mice. Methods: HT22 cells, which were replaced with sugar-free medium, were cultured in a hypoxic incubator for 6 h, and then transferred to a normoxic incubator for reoxygenation for 18 h. The morphological changes of cells were observed, the expression of apoptosis-related factor protein was detected, and the expression of apoptosis-related factor mRNA was detected. The plasmids (sh-nc, sh-cntf, oe-cntf) and inhibitors (sh-akt) were transfected into HT22 cells, and the sugar-free medium was replaced and cultured in the hypoxic incubator for 6 h, then transferred to a normal-oxygen incubator for 18h. The expression of HT22 apoptosis-related factors and AKT/STAT3 protein in each group were detected. Results: Compared with the normal group, the apoptosis level of the OGD treatment group was increased and the anti-apoptosis ability was decreased. Under the microscope, the cell body vacuoles of the OGD group were significantly increased. Compared with the normal group, the apoptosis level of the OGD group and the sh-nc+OGD group was significantly increased, and there was no significant difference between the two groups. The apoptosis level of sh-cntf group was further increased, but the apoptosis level of oe-cntf group was significantly decreased. Compared with the normal group, the apoptosis level in the AKT knockdown group (sh-akt) and the co-knockdown group with AKT and CNTF (sh-akt+cntf) was significantly increased, and the apoptosis level in the sh-akt+cntf group was more significantly increased. Apoptosis was inhibited after AKT knockdown and CNTF overexpression (sh-akt+oecntf). Conclusion: CNTF plays a protective role by up-regulating AKT/STAT3 signaling pathway and activating phosphorylation at phosphoric acid site 705 in STAT3, thereby inhibiting the apoptosis of neuronal cells after OGD.
[1] Ermawati S, Triniputvi WY, Ilyas MF, et al. Research on neurotrophic factor for glaucoma: a worldwide bibliometric analysis[J]. Int J Ophthalmol, 2025, 18(9): 1747-1758.
[2] Rafii P, Cruz PR, Ettich J, et al. Engineered interleukin-6-derived cytokines recruit artificial receptor complexes and disclose CNTF signaling via the OSMR[J]. J Biol Chem, 2024, 300(5): 107251.
[3] Zhou B, Mu K, Yu X, et al. Serum Levels and Clinical Significance of NSE, BDNF and CNTF in Patients with Cancer-associated Ischemic Stroke Complicated with Post-stroke Depression[J]. Actas Esp Psiquiatr, 2024, 52(4): 474-483.
[4] Hu Z, Deng N, Liu K, et al. CNTF-STAT3-IL-6 Axis Mediates Neuroinflammatory Cascade across Schwann Cell-Neuron-Microglia[J]. Cell Rep, 2020, 31(7): 107657.
[5] Liu H, Liu G, Bi Y. CNTF regulates neurite outgrowth and neuronal migration through JAK2/STAT3 and PI3K/Akt signaling pathways of DRG explants with gp120-induced neurotoxicity in vitro[J]. Neurosci Lett, 2014, 569: 110-115.
[6] Li W, Wei D, Zhu Z, et al. Dl-3-n-Butylphthalide Alleviates Hippocampal Neuron Damage in Chronic Cerebral Hypoperfusion via Regulation of the CNTF/CNTFRα/JAK2/STAT3 Signaling Pathways[J]. Front Aging Neurosci, 2020, 12: 587403.
[7] Gu YL, Gao GQ, Ma N, et al. CNTF protects neurons from hypoxic injury through the activation of STAT3pTyr705[J]. Int J Mol Med, 2016, 38(6): 1915-1921.
[8] Heo JH, Yoon JA, Ahn EK, et al. Intraperitoneal administration of adipose tissue-derived stem cells for the rescue of retinal degeneration in a mouse model via indigenous CNTF up-regulation by IL-6[J]. J Tissue Eng Regen Med, 2018, 12(3): e1370-e1382.
[9] Lv L, Qian J, Sang J, et al. Protective effects of PIK3CG knockdown against OGD/R-induced neuronal damage via inhibition of autophagy through the AMPK/mTOR pathway[J]. Neuroscience, 2025, 565: 91-98.
[10] Zhang W, Lu J, Gao Y, et al. NRIP1 is a downstream target of YY1 in promoting OGD/R-induced H9c2 cardiomyocyte injury and mitochondrial dysfunction[J]. Histol Histopathol, 2024: 18820.
[11] Wu L, Cheng Y, Wang R, et al. NDRG2 regulates glucose metabolism and ferroptosis of OGD/R-treated astrocytes by the Wnt/β-catenin signaling[J]. J Biochem Mol Toxicol, 2024, 38(9): e23827.
[12] Zheng K, Zhang Q, Sheng Z, et al. Ciliary Neurotrophic Factor (CNTF) Protects Myocardial Cells from Oxygen Glucose Deprivation (OGD)/Re-Oxygenation via Activation of Akt-Nrf2 Signaling[J]. Cell Physiol Biochem, 2018, 51(4): 1852-1862.
[13] Zhang T, Liu W, Yang J, et al. Components of Salvia miltiorrhiza and Panax notoginseng Protect Pericytes Against OGD/R-Induced Injury via Regulating the PI3K/AKT/mTOR and JNK/ERK/P38 Signaling Pathways[J]. J Mol Neurosci, 2022, 72(12): 2377-2388.
[14] Askarian-amiri S, Fotovat eskandari H, ramezani F, et al. Lavender protects H9c2 cardiomyocytes against oxygen-glucose deprivation (OGD)-induced injury via targeting the JAK2/STAT3 pathway[J]. Iran J Basic Med Sci, 2022, 25(2): 263-267.
[15] Yuan J, Zeng L, Sun Y, et al. SH2B1 protects against OGD/R-induced apoptosis in PC12 cells via activation of the JAK2/STAT3 signaling pathway[J]. Mol Med Rep, 2018, 18(3): 2613-2620.
