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Effect of MALT1 inhibition by MI‑2 on the microglial phenotype switch, inflammatory cytokine secretion, neuronal loss and oxidative stress in Alzheimer's disease

  • Authors:
    • Yuanlong Li
    • Hua Fan
    • Xiong Han
    • Ming Ni
    • Xiaodan Hou
    • Hailan Xia
    • Yinzhu Shi
    • Lin Zhang
    • Jun Sun
  • View Affiliations / Copyright

    Affiliations: Department of Pharmacy, Henan Provincial People's Hospital, Zhengzhou, Henan 450003, P.R. China, School of Clinical Medicine, The First Affiliated Hospital of Henan University of Science and Technology, Henan University of Science and Technology, Luoyang, Henan 471003, P.R. China, Department of Neurology, Henan Provincial People's Hospital, Zhengzhou, Henan 450003, P.R. China, Department of Clinical Pharmacy, Fuwai Central China Cardiovascular Hospital, Zhengzhou, Henan 450053, P.R. China, Ward of Heart Failure, Fuwai Central China Cardiovascular Hospital, Zhengzhou, Henan 450000, P.R. China, Department of Pharmacy, Henan Province Reproductive Maternity Hospital, Zhengzhou, Henan 450000, P.R. China
    Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 302
    |
    Published online on: August 31, 2026
       https://doi.org/10.3892/ijmm.2026.5973
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Abstract

Mucosa‑associated lymphoid tissue lymphoma translocation protein 1 (MALT1) is a key paracaspase enzyme regulating immune responses, inflammation and oxidative stress. The present study aimed to investigate the effect of MALT1 inhibition on neuroinflammation, neuronal loss and oxidative stress in Alzheimer's disease (AD). A co‑culture system involving microglia and neuron cells under β‑amyloid (Aβ) intervention was used to establish AD cellular models using human microglia HMC3 cells and neuroblastoma SH‑SY5Y cells and mouse microglia BV‑2 and hippocampal neuron HT‑22 cells. The inhibition of MALT1 proteolytic activity was achieved by MALT1 inhibitor 2 (MI‑2) treatment, and the NF‑κB pathway was activated by phorbol 12‑myristate 13‑acetate (PMA) treatment in HMC3 and BV‑2 cells. Western blotting, ELISA, Cell Counting Kit‑8, EdU staining, reactive oxygen species (ROS) detection and reduced glutathione (GSH) assays were performed to evaluate molecular changes, inflammatory responses, neuronal viability and oxidative stress. MALT1 expression was upregulated following Aβ treatment in HMC3 and BV‑2 cells. MALT1 inhibition by MI‑2 suppressed the microglial M1 phenotype but enhanced the M2 phenotype, reduced the levels of the proinflammatory cytokines TNF‑α and IL‑1β and inactivated the NF‑κB pathway in HMC3 and BV‑2 cells. Moreover, microglial MALT1 inhibition elevated cell viability (verified by Cell Counting Kit‑8 and EdU assays), increased the level of reduced glutathione and decreased the levels of reactive oxygen species in SH‑SY5Y and HT‑22 cells. NF‑κB activation by PMA attenuated the effects of MALT1 inhibition on the microglial phenotype switch and proinflammatory cytokine secretion in HMC3 and BV‑2 cells, as well as cell viability and oxidative stress in SH‑SY5Y and HT‑22 cells. The present study reveals that MALT1 inhibition may suppress microglial M1 phenotype, neuroinflammation, neuronal loss and oxidative stress by inactivating the NF‑κB pathway in AD.
View Figures

Figure 1

Co-culture system design. Aβ,
amyloid-β; MI-2, MALT1 inhibitor-2; PMA, phorbol 12-myristate
13-acetate; CM, culture medium; CCK, Cell Counting Kit; ROS,
reactive oxygen species; GSH, glutathione.

Figure 2

Effect of MALT1 inhibition on the
microglial phenotype switch and inflammation. Expression levels of
(A) MALT1 and CYLD, (B) microglial M1 phenotype marker iNOS and the
M2 phenotype marker ARG1 and (C) inflammatory cytokines TNF-α and
IL-1β in HMC3 and BV-2 cells. *P<0.05,
**P<0.01, ***P<0.001. ns, not
significant; MALT, mucosa-associated lymphoid tissue lymphoma
translocation protein; CYLD, CYLD lysine 63 deubiquitinase; iNOS,
inducible nitric oxide synthase; ARG, arginase; Aβ, amyloid-β;
MI-2, MALT1 inhibitor-2.

Figure 3

Effect of MALT1 inhibition on the
NF-κB pathway. Expression of p-p65 in (A) HMC3 and (B) BV-2 cells.
*P<0.05, **P<0.01. MALT,
mucosa-associated lymphoid tissue lymphoma translocation protein;
p-, phosphorylated; Aβ, amyloid-β; MI-2, MALT1 inhibitor-2.

Figure 4

Effect of microglial MALT1 inhibition
on neuronal viability and oxidative stress. (A) OD values obtained
from Cell Counting Kit-8 assay. (B) EdU-positive cells. Relative
(C) ROS and (D) GSH levels in SH-SY5Y and HT-22 cells.
*P<0.05, **P<0.01,
***P<0.001. ns, not significant; MALT,
mucosa-associated lymphoid tissue lymphoma translocation protein;
OD, optical density; ROS, reactive oxygen species; GSH,
glutathione; Aβ, amyloid-β; MI-2, MALT1 inhibitor-2.

Figure 5

Effect of PMA on the NF-κB pathway.
Expression of p-p65 in (A) HMC3 cells and (B) BV-2 cells.
*P<0.05, **P<0.01,
***P<0.001. PMA, phorbol 12-myristate 13-acetate; p-,
phosphorylated; Aβ, amyloid-β; MI-2, MALT1 inhibitor-2.

Figure 6

Effect of NF-κB activation on the
microglial phenotype switch and inflammation. Expression levels of
(A) microglial M1 phenotype marker iNOS and the M2 phenotype marker
ARG1 and (B) inflammatory cytokines TNF-α and IL-1β in HMC3 and
BV-2 cells. *P<0.05, **P<0.01,
***P<0.001. ns, not significant; iNOS, inducible
nitric oxide synthase; ARG, arginase; Aβ, amyloid-β; MI-2, MALT1
inhibitor-2.

Figure 7

Effect of microglial NF-κB activation
on neuronal viability and oxidative stress. (A) OD values obtained
from Cell Counting Kit-8 assay. (B) EdU-positive cells. Relative
(C) ROS and (D) GSH levels in SH-SY5Y and HT-22 cells.
*P<0.05, **P<0.01,
***P<0.001; OD, optical density; ROS, reactive oxygen
species; GSH, glutathione; Aβ, amyloid-β; MI-2, MALT1 inhibitor-2;
PMA, phorbol 12-myristate 13-acetate; ns, not significant.
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Copy and paste a formatted citation
Spandidos Publications style
Li Y, Fan H, Han X, Ni M, Hou X, Xia H, Shi Y, Zhang L and Sun J: Effect of MALT1 inhibition by MI‑2 on the microglial phenotype switch, inflammatory cytokine secretion, neuronal loss and oxidative stress in Alzheimer's disease. Int J Mol Med 58: 302, 2026.
APA
Li, Y., Fan, H., Han, X., Ni, M., Hou, X., Xia, H. ... Sun, J. (2026). Effect of MALT1 inhibition by MI‑2 on the microglial phenotype switch, inflammatory cytokine secretion, neuronal loss and oxidative stress in Alzheimer's disease. International Journal of Molecular Medicine, 58, 302. https://doi.org/10.3892/ijmm.2026.5973
MLA
Li, Y., Fan, H., Han, X., Ni, M., Hou, X., Xia, H., Shi, Y., Zhang, L., Sun, J."Effect of MALT1 inhibition by MI‑2 on the microglial phenotype switch, inflammatory cytokine secretion, neuronal loss and oxidative stress in Alzheimer's disease". International Journal of Molecular Medicine 58.5 (2026): 302.
Chicago
Li, Y., Fan, H., Han, X., Ni, M., Hou, X., Xia, H., Shi, Y., Zhang, L., Sun, J."Effect of MALT1 inhibition by MI‑2 on the microglial phenotype switch, inflammatory cytokine secretion, neuronal loss and oxidative stress in Alzheimer's disease". International Journal of Molecular Medicine 58, no. 5 (2026): 302. https://doi.org/10.3892/ijmm.2026.5973
Copy and paste a formatted citation
x
Spandidos Publications style
Li Y, Fan H, Han X, Ni M, Hou X, Xia H, Shi Y, Zhang L and Sun J: Effect of MALT1 inhibition by MI‑2 on the microglial phenotype switch, inflammatory cytokine secretion, neuronal loss and oxidative stress in Alzheimer's disease. Int J Mol Med 58: 302, 2026.
APA
Li, Y., Fan, H., Han, X., Ni, M., Hou, X., Xia, H. ... Sun, J. (2026). Effect of MALT1 inhibition by MI‑2 on the microglial phenotype switch, inflammatory cytokine secretion, neuronal loss and oxidative stress in Alzheimer's disease. International Journal of Molecular Medicine, 58, 302. https://doi.org/10.3892/ijmm.2026.5973
MLA
Li, Y., Fan, H., Han, X., Ni, M., Hou, X., Xia, H., Shi, Y., Zhang, L., Sun, J."Effect of MALT1 inhibition by MI‑2 on the microglial phenotype switch, inflammatory cytokine secretion, neuronal loss and oxidative stress in Alzheimer's disease". International Journal of Molecular Medicine 58.5 (2026): 302.
Chicago
Li, Y., Fan, H., Han, X., Ni, M., Hou, X., Xia, H., Shi, Y., Zhang, L., Sun, J."Effect of MALT1 inhibition by MI‑2 on the microglial phenotype switch, inflammatory cytokine secretion, neuronal loss and oxidative stress in Alzheimer's disease". International Journal of Molecular Medicine 58, no. 5 (2026): 302. https://doi.org/10.3892/ijmm.2026.5973
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