DOCK8 interference alleviates Aβ‑induced damage of BV2 cells by inhibiting STAT3/NLRP3/NF‑κB signaling

Zhou,Xueying; Hu,Ji; Xu,Deyi; Zhang,Sheng; Wang,Qianyan

doi:10.3892/etm.2023.11833

DOCK8 interference alleviates Aβ‑induced damage of BV2 cells by inhibiting STAT3/NLRP3/NF‑κB signaling

Authors:
- Xueying Zhou
- Ji Hu
- Deyi Xu
- Sheng Zhang
- Qianyan Wang
View Affiliations

Affiliations: Department of Psychiatry, Liyuan Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430077, P.R. China, Department of Anesthesiology, Liyuan Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430077, P.R. China, Department of Cardiology, Liyuan Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430077, P.R. China
Published online on: February 10, 2023 https://doi.org/10.3892/etm.2023.11833
Article Number: 134
Copyright: © Zhou et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Dementia is defined as memory loss and other cognitive decline and it severely influences daily life. Alzheimer's disease (AD) is the most common cause of dementia. Dedicator of cytokinesis 8 (DOCK8) is reported to be involved in neurological diseases. The present study focused on investigating the role that DOCK8 serves in AD and addressing its hidden regulatory mechanism. Initially, Aβ_1‑42 (Aβ) was applied for the administration of BV2 cells. Subsequently, the mRNA and protein expression levels of DOCK8 were evaluated utilizing reverse transcription‑quantitative PCR (RT‑qPCR) and western blotting. After the DOCK8 silencing, immunofluorescence staining (IF), ELISA, wound healing and Transwell assays were applied to assess ionized calcium binding adapter molecule‑1 (IBA‑1) expression, release of inflammatory factors, migration and invasion in Aβ‑induced BV2 cells. IF was used to evaluate cluster of differentiation (CD)11b expression. RT‑qPCR and western blotting were to analyze the levels of M1 cell markers inducible nitric oxide synthase (iNOS) and CD86. The expression of STAT3/NLR family pyrin domain containing 3 (NLRP3)/NF‑κB signaling‑related proteins were determined by western blotting. Finally, the viability and apoptosis in hippocampal HT22 cells with DOCK8 depletion were estimated. Results revealed that Aβ induction greatly stimulated the expression levels of IBA‑1 and DOCK8. DOCK8 silencing suppressed Aβ‑induced inflammation, migration and invasion of BV2 cells. Additionally, DOCK8 deficiency conspicuously decreased the expression levels of CD11b, iNOS and CD86. The expression of phosphorylated (p‑)STAT3, NLRP3, ASC, caspase1 and p‑p65 was downregulated in Aβ‑induced BV2 cells after DOCK8 depletion. STAT3 activator Colivelin reversed the effects of DOCK8 knockdown on IBA‑1 expression, inflammation, migration, invasion and M1 cell polarization. In addition, the viability and apoptosis in hippocampal HT22 cells stimulated by neuroinflammatory release of BV2 cells were repressed following DOCK8 deletion. Collectively, DOCK8 interference alleviated Aβ‑induced damage of BV2 cells by inhibiting STAT3/NLRP3/NF‑κB signaling.

View Figures

View References

1	Cui Y, Wang Y, Zhao D, Feng X, Zhang L and Liu C: Loganin prevents BV-2 microglia cells from Abeta1-42 -induced inflammation via regulating TLR4/TRAF6/NF-κB axis. Cell Biol Int. 42:1632–1642. 2018.PubMed/NCBI View Article : Google Scholar
2	Lane CA, Hardy J and Schott JM: Alzheimer's disease. Eur J Neurol. 25:59–70. 2018.PubMed/NCBI View Article : Google Scholar
3	De-Paula VJ, Radanovic M, Diniz BS and Forlenza OV: Alzheimer's disease. Subcell Biochem. 65:329–352. 2012.PubMed/NCBI View Article : Google Scholar
4	Calsolaro V and Edison P: Neuroinflammation in Alzheimer's disease: Current evidence and future directions. Alzheimers Dement. 12:719–732. 2016.PubMed/NCBI View Article : Google Scholar
5	Varnum MM and Ikezu T: The classification of microglial activation phenotypes on neurodegeneration and regeneration in Alzheimer's disease brain. Arch Immunol Ther Exp (Warsz). 60:251–266. 2012.PubMed/NCBI View Article : Google Scholar
6	Unger MS, Li E, Scharnagl L, Poupardin R, Altendorfer B, Mrowetz H, Hutter-Paier B, Weiger TM, Heneka MT, Attems J and Aigner L: CD8(+) T-cells infiltrate Alzheimer's disease brains and regulate neuronal- and synapse-related gene expression in APP-PS1 transgenic mice. Brain Behav Immun. 89:67–86. 2020.PubMed/NCBI View Article : Google Scholar
7	Mosher KI and Wyss-Coray T: Microglial dysfunction in brain aging and Alzheimer's disease. Biochem Pharmacol. 88:594–604. 2014.PubMed/NCBI View Article : Google Scholar
8	Kang SY, Jung HW, Lee MY, Lee HW, Chae SW and Park YK: Effect of the semen extract of Cuscuta chinensis on inflammatory responses in LPS-stimulated BV-2 microglia. Chin J Nat Med. 12:573–581. 2014.PubMed/NCBI View Article : Google Scholar
9	Namekata K, Kimura A, Kawamura K, Harada C and Harada T: Dock GEFs and their therapeutic potential: Neuroprotection and axon regeneration. Prog Retin Eye Res. 43:1–16. 2014.PubMed/NCBI View Article : Google Scholar
10	Kearney CJ, Randall KL and Oliaro J: DOCK8 regulates signal transduction events to control immunity. Cell Mol Immunol. 14:406–411. 2017.PubMed/NCBI View Article : Google Scholar
11	Namekata K, Guo X, Kimura A, Arai N, Harada C and Harada T: DOCK8 is expressed in microglia, and it regulates microglial activity during neurodegeneration in murine disease models. J Biol Chem. 294:13421–13433. 2019.PubMed/NCBI View Article : Google Scholar
12	Biggs CM, Keles S and Chatila TA: DOCK8 deficiency: Insights into pathophysiology, clinical features and management. Clin Immunol. 181:75–82. 2017.PubMed/NCBI View Article : Google Scholar
13	Moon MY, Kim HJ, Li Y, Kim JG, Jeon YJ, Won HY, Kim JS, Kwon HY, Choi IG, Ro E, et al: Involvement of small GTPase RhoA in the regulation of superoxide production in BV2 cells in response to fibrillar Abeta peptides. Cell Signal. 25:1861–1869. 2013.PubMed/NCBI View Article : Google Scholar
14	Cimino PJ, Yang Y, Li X, Hemingway JF, Cherne MK, Khademi SB, Fukui Y, Montine KS, Montine TJ and Keene CD: Ablation of the microglial protein DOCK2 reduces amyloid burden in a mouse model of Alzheimer's disease. Exp Mol Pathol. 94:366–371. 2013.PubMed/NCBI View Article : Google Scholar
15	Jiang CQ, Ma LL, Lv ZD, Feng F, Chen Z and Liu ZD: Polydatin induces apoptosis and autophagy via STAT3 signaling in human osteosarcoma MG-63 cells. J Nat Med. 74:533–544. 2020.PubMed/NCBI View Article : Google Scholar
16	Jian M, Kwan JS, Bunting M, Ng RC and Chan KH: Adiponectin suppresses amyloid-β oligomer (AβO)-induced inflammatory response of microglia via AdipoR1-AMPK-NF-κB signaling pathway. J Neuroinflammation. 16(110)2019.PubMed/NCBI View Article : Google Scholar
17	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
18	Latta CH, Brothers HM and Wilcock DM: Neuroinflammation in Alzheimer's disease; A source of heterogeneity and target for personalized therapy. Neuroscience. 302:103–111. 2015.PubMed/NCBI View Article : Google Scholar
19	Heneka MT, Carson MJ, El Khoury J, Landreth GE, Brosseron F, Feinstein DL, Jacobs AH, Wyss-Coray T, Vitorica J, Ransohoff RM, et al: Neuroinflammation in Alzheimer's disease. Lancet Neurol. 14:388–405. 2015.PubMed/NCBI View Article : Google Scholar
20	Heppner FL, Ransohoff RM and Becher B: Immune attack: The role of inflammation in Alzheimer disease. Nat Rev Neurosci. 16:358–372. 2015.PubMed/NCBI View Article : Google Scholar
21	Imai Y, Ibata I, Ito D, Ohsawa K and Kohsaka S: A novel gene iba1 in the major histocompatibility complex class III region encoding an EF hand protein expressed in a monocytic lineage. Biochem Biophys Res Commun. 224:855–862. 1996.PubMed/NCBI View Article : Google Scholar
22	Maneu V, Noailles A, Megias J, Gómez-Vicente V, Carpena N, Gil ML, Gozalbo D and Cuenca N: Retinal microglia are activated by systemic fungal infection. Invest Ophthalmol Vis Sci. 55:3578–3585. 2014.PubMed/NCBI View Article : Google Scholar
23	Shi FJ, Xie H, Zhang CY, Qin HF, Zeng XW, Lou H, Zhang L, Xu GT, Zhang JF and Xu GX: Is Iba-1 protein expression a sensitive marker for microglia activation in experimental diabetic retinopathy? Int J Ophthalmol. 14:200–208. 2021.PubMed/NCBI View Article : Google Scholar
24	Muhammad T, Ikram M, Ullah R, Rehman SU and Kim MO: Hesperetin, a citrus flavonoid, attenuates LPS-induced neuroinflammation, apoptosis and memory impairments by modulating TLR4/NF-κB signaling. Nutrients. 11(648)2019.PubMed/NCBI View Article : Google Scholar
25	Park T, Chen H, Kevala K, Lee JW and Kim HY: N-Docosahexaenoylethanolamine ameliorates LPS-induced neuroinflammation via cAMP/PKA-dependent signaling. J Neuroinflammation. 13(284)2016.PubMed/NCBI View Article : Google Scholar
26	Zhu SH, Liu BQ, Hao MJ, Fan YX, Qian C, Teng P, Zhou XW, Hu L, Liu WT, Yuan ZL and Li QP: Paeoniflorin suppressed high glucose-induced retinal microglia MMP-9 expression and inflammatory response via inhibition of TLR4/NF-κB pathway through upregulation of SOCS3 in diabetic retinopathy. Inflammation. 40:1475–1486. 2017.PubMed/NCBI View Article : Google Scholar
27	Hopperton KE, Mohammad D, Trepanier MO, Giuliano V and Bazinet RP: Markers of microglia in post-mortem brain samples from patients with Alzheimer's disease: A systematic review. Mol Psychiatry. 23:177–198. 2018.PubMed/NCBI View Article : Google Scholar
28	Dobryakova YV, Kasianov A, Zaichenko MI, Stepanichev MY, Chesnokova EA, Kolosov PM, Markevich VA and Bolshakov AP: Intracerebroventricular administration of (192)IgG-Saporin alters expression of microglia-associated genes in the dorsal but not ventral hippocampus. Front Mol Neurosci. 10(429)2017.PubMed/NCBI View Article : Google Scholar
29	Aydin SE, Kilic SS, Aytekin C, Kumar A, Porras O, Kainulainen L, Kostyuchenko L, Genel F, Kütükcüler N, Karaca N, et al: DOCK8 deficiency: Clinical and immunological phenotype and treatment options-a review of 136 patients. J Clin Immunol. 35:189–198. 2015.PubMed/NCBI View Article : Google Scholar
30	Zhang Z, Bao Y, Zhou L, Ye Y, Fu W and Sun C: DOCK8 Serves as a prognostic biomarker and is related to immune infiltration in patients with HPV positive head and neck squamous cell carcinoma. Cancer Control. 28(10732748211011951)2021.PubMed/NCBI View Article : Google Scholar
31	Xu X, Han L, Zhao G, Xue S, Gao Y, Xiao J, Zhang S, Chen P, Wu ZY, Ding J, et al: LRCH1 interferes with DOCK8-Cdc42-induced T cell migration and ameliorates experimental autoimmune encephalomyelitis. J Exp Med. 214:209–226. 2017.PubMed/NCBI View Article : Google Scholar
32	Miyamoto Y, Torii T, Kawahara K, Tanoue A and Yamauchi J: Dock8 interacts with Nck1 in mediating Schwann cell precursor migration. Biochem Biophys Rep. 6:113–123. 2016.PubMed/NCBI View Article : Google Scholar
33	Ham H, Guerrier S, Kim J, Schoon RA, Anderson EL, Hamann MJ, Lou Z and Billadeau DD: Dedicator of cytokinesis 8 interacts with talin and Wiskott-Aldrich syndrome protein to regulate NK cell cytotoxicity. J Immunol. 190:3661–3669. 2013.PubMed/NCBI View Article : Google Scholar
34	Hillmer EJ, Zhang H, Li HS and Watowich SS: STAT3 signaling in immunity. Cytokine Growth Factor Rev. 31:1–15. 2016.PubMed/NCBI View Article : Google Scholar
35	Takeda K, Kaisho T, Yoshida N, Takeda J, Kishimoto T and Akira S: Stat3 activation is responsible for IL-6-dependent T cell proliferation through preventing apoptosis: Generation and characterization of T cell-specific Stat3-deficient mice. J Immunol. 161:4652–4660. 1998.PubMed/NCBI
36	Kortylewski M and Yu H: Role of Stat3 in suppressing anti-tumor immunity. Curr Opin Immunol. 20:228–233. 2008.PubMed/NCBI View Article : Google Scholar
37	Egwuagu CE: STAT3 in CD4⁺ T helper cell differentiation and inflammatory diseases. Cytokine. 47:149–156. 2009.PubMed/NCBI View Article : Google Scholar
38	Keles S, Charbonnier LM, Kabaleeswaran V, Reisli I, Genel F, Gulez N, Al-Herz W, Ramesh N, Perez-Atayde A, Karaca NE, et al: Dedicator of cytokinesis 8 regulates signal transducer and activator of transcription 3 activation and promotes TH17 cell differentiation. J Allergy Clin Immunol. 138:1384–1394 e1382. 2016.PubMed/NCBI View Article : Google Scholar
39	Zhu H, Jian Z, Zhong Y, Ye Y, Zhang Y, Hu X, Pu B, Gu L and Xiong X: Janus kinase inhibition ameliorates ischemic stroke injury and neuroinflammation through reducing NLRP3 inflammasome activation via JAK2/STAT3 pathway inhibition. Front Immunol. 12(714943)2021.PubMed/NCBI View Article : Google Scholar
40	Dai R, Jiang Q, Zhou Y, Lin R, Lin H, Zhang Y, Zhang J and Gao X: Lnc-STYK1-2 regulates bladder cancer cell proliferation, migration, and invasion by targeting miR-146b-5p expression and AKT/STAT3/NF-kB signaling. Cancer Cell Int. 21(408)2021.PubMed/NCBI View Article : Google Scholar
41	Pal M, Bao W, Wang R, Liu Y, An X, Mitchell WB, Lobo CA, Minniti C, Shi PA, Manwani D, et al: Hemolysis inhibits humoral B-cell responses and modulates alloimmunization risk in patients with sickle cell disease. Blood. 137:269–280. 2021.PubMed/NCBI View Article : Google Scholar