This study explores the neuroprotective effects of Stanniocalcin-1 (STC1) in the context of ischemic stroke, a major cause of death and disability worldwide. Although STC1 is recognized for its cytoprotective properties, its role in cerebral ischemia/reperfusion (I/R) injury has not been fully characterized. Our objective was to clarify the effects and underlying mechanisms of STC1 using a mouse model of middle cerebral artery occlusion/reperfusion (MCAO/R). We administered recombinant human STC1 (rh-STC1) intranasally and assessed cerebral injury through various methods including neurological scoring, behavioral tests, and TUNEL/Nissl staining. Additionally, we evaluated markers of oxidative stress and ferroptosis. The results demonstrated that endogenous STC1 levels were decreased following ischemia. Treatment with rh-STC1 led to significant improvements in neurological function, reduction in infarct volume, and decreased permeability of the blood–brain barrier. Furthermore, rh-STC1 treatment mitigated neuronal apoptosis, oxidative stress, and ferroptosis. Mechanistically, rh-STC1 was found to interact with calreticulin (CRT), activating the CAMKK2/AMPK/GSK-3β/Nrf2 signaling pathway. Notably, the neuroprotective effects of rh-STC1 were abolished by an AMPK inhibitor. In conclusion, our findings indicate that rh-STC1 protects against cerebral I/R injury by inhibiting apoptosis and ferroptosis, primarily via the CRT/CAMKK2/AMPK/GSK-3β/Nrf2 pathway. This positions STC1 as a promising therapeutic target for ischemic stroke.
GBD 2019 Diseases and Injuries Collaborators. Global burden of 369 diseases and injuries in 204 countries and territories, 1990–2019: a systematic analysis for the Global Burden of Disease Study 2019. Lancet2020; 396: 1204–1222.
2.
PuigBBrennaSMagnusT.Molecular communication of a dying neuron in stroke. Int J Mol Sci2018; 19: 2834.
3.
LebesgueDChevaleyreVZukinRS, et al. Estradiol rescues neurons from global ischemia-induced cell death: multiple cellular pathways of neuroprotection. Steroids2009; 74: 555–561.
4.
YaghiSWilleyJZCucchiaraB, et al. Treatment and outcome of hemorrhagic transformation after intravenous alteplase in acute ischemic stroke: a scientific statement for healthcare professionals from the American Heart Association/American Stroke Association. Stroke2017; 48: e343–e361.
5.
MunichSAVakhariaKLevyEI.Overview of mechanical thrombectomy techniques. Neurosurgery2019; 85: S60–S67.
6.
XuHShenJXiaoJ, et al. Neuroprotective effect of cajaninstilbene acid against cerebral ischemia and reperfusion damages by activating AMPK/Nrf2 pathway. J Adv Res2021; 34: 199–210.
7.
YuLZhangYChenQ, et al. Formononetin protects against inflammation associated with cerebral ischemia–reperfusion injury in rats by targeting the JAK2/STAT3 signaling pathway. Biomed Pharmacother2022; 149: 112836.
8.
OjoOBAmooZASaliuIO, et al. Neurotherapeutic potential of kolaviron on neurotransmitter dysregulation, excitotoxicity, mitochondrial electron transport chain dysfunction and redox imbalance in 2-VO brain ischemia/reperfusion injury. Biomed Pharmacother2019; 111: 859–872.
9.
IshibashiKImaiM.Prospect of a stanniocalcin endocrine/paracrine system in mammals. Am J Physiol Renal Physiol2002; 282: F367–F375.
10.
PanJSHuangLBelousovaT, et al. Stanniocalcin-1 inhibits renal ischemia/reperfusion injury via an AMP-activated protein kinase-dependent pathway. J Am Soc Nephrol2015; 26: 364–378.
11.
WangPLiXCaoZ.STC1 ameliorates cognitive impairment and neuroinflammation of Alzheimer’s disease mice via inhibition of ERK1/2 pathway. Immunobiology2021; 226: 152092.
12.
LeungCCWongCK.Effects of STC1 overexpression on tumorigenicity and metabolism of hepatocellular carcinoma. Oncotarget2018; 9: 6852–6861.
13.
JiayanXYaqiMManJ, et al. Epithelial expression and role of secreted STC1 on asthma airway hyperresponsiveness through calcium channel modulation. Allergy2020; 76: 2475–2487.
14.
GaoCFZhangGHYeZH, et al. Usability of serum stanniocalcin-1 as a prognostic biochemical marker of acute supratentorial intracerebral hemorrhage: a prospective cohort study. Int J Gen Med2023; 16: 2791–2803.
15.
ZhangYShanPSrivastavaA, et al. Endothelial stanniocalcin 1 maintains mitochondrial bioenergetics and prevents oxidant-induced lung injury via Toll-like receptor 4. Antioxid Redox Signal2019; 30: 1775–1796.
16.
GuoFLiYWangJ, et al. Stanniocalcin1 (STC1) inhibits cell proliferation and invasion of cervical cancer cells. PLoS One2013; 8: e53989.
17.
BaiYXiaoYDaiY, et al. Stanniocalcin 1 promotes cell proliferation via cyclin E1/cyclin‑dependent kinase 2 in human prostate carcinoma. Oncol Rep2017; 37: 2465–2471.
18.
SonnichsenBFullekrugJNguyenVP, et al. Retention and retrieval: both mechanisms cooperate to maintain calreticulin in the endoplasmic reticulum. J Cell Sci1994; 107: 2705–2717.
19.
LuoMWangXWuS, et al. A20 promotes colorectal cancer immune evasion by upregulating STC1 expression to block “eat-me” signal. Signal Transduct Target Ther2023; 8: 312.
20.
SunBHeSLiuB, et al. Stanniocalcin-1 protected astrocytes from hypoxic damage through the AMPK pathway. Neurochem Res2021; 46: 2948–2957.
21.
WoodsAJohnstoneSRDickersonK, et al. LKB1 is the upstream kinase in the AMP-activated protein kinase cascade. Curr Biol2003; 13: 2004–2008.
22.
WoodsADickersonKHeathR, et al. Ca2+/calmodulin-dependent protein kinase kinase-β acts upstream of AMP-activated protein kinase in mammalian cells. Cell Metab2005; 2: 21–33.
23.
TianXWangYYuanM, et al. Heme oxygenase-1-modified BMMSCs activate AMPK–Nrf2–FTH1 to reduce severe steatotic liver ischemia–reperfusion injury. Digest Dis Sci2023; 68: 4196–4211.
24.
SunXOuZChenR, et al. Activation of the p62–Keap1–NRF2 pathway protects against ferroptosis in hepatocellular carcinoma cells. Hepatology2016; 63: 173–184.
25.
HouWYaoJLiuJ, et al. USP14 inhibition promotes recovery by protecting BBB integrity and attenuating neuroinflammation in MCAO mice. CNS Neurosci Ther2023; 29: 3612–3623.
MaYLiuCRenL, et al. β-1,4-Galactosyltransferase 1 protects against cerebral ischemia injury in mice by suppressing ferroptosis via the TAZ/Nrf2/HO-1 signaling pathway. CNS Neurosci Ther2024; 30: e70030.
28.
ShiXXuLDoychevaDM, et al. Sestrin2, as a negative feedback regulator of mTOR, provides neuroprotection by activation AMPK phosphorylation in neonatal hypoxic–ischemic encephalopathy in rat pups. J Cereb Blood Flow Metab2017; 37: 1447–1460.
29.
YeYJianZJinT, et al. NOX2-mediated reactive oxygen species are double-edged swords in focal cerebral ischemia in mice. J Neuroinflammation2022; 19: 184.
30.
LiYDongYHeY, et al. Endothelial cell-based vascular bandages for blood–brain barrier repair and targeted siRNA delivery. Adv Sci2026; 13: e17780.
31.
ChenYVeenmanLLiaoM, et al. Enhanced angiogenesis in the thalamus induced by a novel TSPO ligand ameliorates cognitive deficits after focal cortical infarction. J Cereb Blood Flow Metab2024; 44: 477–490.
ZhouSLiuCWangJ, et al. CCL5 mediated astrocyte-T cell interaction disrupts blood–brain barrier in mice after hemorrhagic stroke. J Cereb Blood Flow Metab2024; 44: 367–383.
36.
MignotteBVayssiereJL.Mitochondria and apoptosis. Eur J Biochem1998; 252: 1–15.
37.
ClelandMMNorrisKLKarbowskiM, et al. Bcl-2 family interaction with the mitochondrial morphogenesis machinery. Cell Death Differ2011; 18: 235–247.
38.
LiT.SLC7A11 in hepatocellular carcinoma: potential mechanisms, regulation, and clinical significance. Am J Cancer Res2024; 14: 2326–2342.
39.
CuiCYangFLiQ.Post-translational modification of GPX4 is a promising target for treating ferroptosis-related diseases. Front Mol Biosci2022; 9: 901565.
40.
MassieABoilléeSHewettS, et al. Main path and byways: non-vesicular glutamate release by system xc(−) as an important modifier of glutamatergic neurotransmission. J Neurochem2015; 135: 1062–1079.
41.
DollSPronethBTyurinaYY, et al. ACSL4 dictates ferroptosis sensitivity by shaping cellular lipid composition. Nat Chem Biol2017; 13: 91–98.
42.
GarciaDShawRJ.AMPK: mechanisms of cellular energy sensing and restoration of metabolic balance. Mol Cell2017; 66: 789–800.
43.
DuanJCuiJYangZ, et al. Neuroprotective effect of Apelin 13 on ischemic stroke by activating AMPK/GSK-3β/Nrf2 signaling. J Neuroinflammation2019; 16: 24.
44.
WangZYaoMJiangL, et al. Dexmedetomidine attenuates myocardial ischemia/reperfusion-induced ferroptosis via AMPK/GSK-3beta/Nrf2 axis. Biomed Pharmacother2022; 154: 113572.
45.
SunXWangDZhangT, et al. Eugenol attenuates cerebral ischemia–reperfusion injury by enhancing autophagy via AMPK–mTOR–P70S6K pathway. Front Pharmacol2020; 11: 84.
46.
LiuYSChangYCKuoWW, et al. Calreticulin nuclear translocalization alleviates CaM/CaMKII/CREB signaling pathway to enhance chemosensitivity in HDAC inhibitor-resistant hepatocellular carcinoma cells. Aging2022; 14: 5097–5115.
47.
McAloonLMMullerAGNayK, et al. CaMKK2: bridging the gap between Ca2+ signaling and energy-sensing. Essays Biochem2024; 68: 309–320.
48.
NajarMARexDModiPK, et al. A complete map of the calcium/calmodulin-dependent protein kinase kinase 2 (CAMKK2) signaling pathway. J Cell Commun Signal2021; 15: 283–290.
49.
WangYRuanWMiJ, et al. Balasubramide derivative 3C modulates microglia activation via CaMKKβ-dependent AMPK/PGC-1α pathway in neuroinflammatory conditions. Brain Behav Immun2018; 67: 101–117.
50.
ChangACJellinekDAReddelRR.Mammalian stanniocalcins and cancer. Endocr Relat Cancer2003; 10: 359–373.
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