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Correlative Effects of PARP12 mRNA Knockdown on Pro-inflammatory Gene Expression and Alternative Splicing in BV2 Microglial Cells.

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Τίτλος: Correlative Effects of PARP12 mRNA Knockdown on Pro-inflammatory Gene Expression and Alternative Splicing in BV2 Microglial Cells.
Συγγραφείς: Hu B; Hubei University of Chinese Medicine, Wuhan, 430065, People's Republic of China.; Department of Orthopedics, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan, 430074, People's Republic of China.; Hubei Shizhen Laboratory, Wuhan, 430074, People's Republic of China.; Department of Orthopedics, Affiliated Hospital of Hubei University of Chinese Medicine, Wuhan, 430074, People's Republic of China., Zhou W; Hubei University of Chinese Medicine, Wuhan, 430065, People's Republic of China.; Department of Orthopedics, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan, 430074, People's Republic of China.; Hubei Shizhen Laboratory, Wuhan, 430074, People's Republic of China.; Department of Orthopedics, Affiliated Hospital of Hubei University of Chinese Medicine, Wuhan, 430074, People's Republic of China., Chen H; Department of Orthopedics, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan, 430074, People's Republic of China. ch1222@126.com.; Hubei Shizhen Laboratory, Wuhan, 430074, People's Republic of China. ch1222@126.com.; Department of Orthopedics, Affiliated Hospital of Hubei University of Chinese Medicine, Wuhan, 430074, People's Republic of China. ch1222@126.com., Li F; Department of Orthopedics, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan, 430074, People's Republic of China. 690683196@qq.com.; Hubei Shizhen Laboratory, Wuhan, 430074, People's Republic of China. 690683196@qq.com.; Department of Orthopedics, Affiliated Hospital of Hubei University of Chinese Medicine, Wuhan, 430074, People's Republic of China. 690683196@qq.com.
Πηγή: Molecular neurobiology [Mol Neurobiol] 2026 Aug 15; Vol. 63 (1). Date of Electronic Publication: 2026 Aug 15.
Τύπος έκδοσης: Journal Article
Γλώσσα: English
Στοιχεία περιοδικού: Publisher: Humana Press Country of Publication: United States NLM ID: 8900963 Publication Model: Electronic Cited Medium: Internet ISSN: 1559-1182 (Electronic) Linking ISSN: 08937648 NLM ISO Abbreviation: Mol Neurobiol Subsets: MEDLINE
Imprint Name(s): Original Publication: Clifton, NJ : Humana Press, c1987-
Ιατρικοί όροι (MeSH): Microglia*/metabolism , Microglia*/pathology , Alternative Splicing*/genetics , RNA, Messenger*/genetics , RNA, Messenger*/metabolism , Inflammation*/genetics , Inflammation*/pathology , Poly(ADP-ribose) Polymerases*/genetics , Poly(ADP-ribose) Polymerases*/metabolism , Inflammation Mediators*/metabolism , Gene Knockdown Techniques* , Gene Expression Regulation*, Transcriptome/genetics ; Animals ; Mice ; Cell Line
Περίληψη: Neuroinflammation is a key mechanism underlying neuropathic pain, in which microglial cells play a central role. Poly(ADP-ribose) polymerase 12 (PARP12) is an interferon-stimulated gene (ISG) containing a predicted RNA-binding domain and may be involved in inflammatory regulation and RNA metabolism; however, its posttranscriptional regulatory functions in microglia remain to be fully elucidated. PARP12 was stably knocked down in BV2 microglial cells using lentiviral shRNA. Quantitative PCR confirmed a significant reduction in mRNA levels (knockdown efficiency 78.3%), but Western blot analysis showed no successful downregulation of PARP12 protein levels. Transcriptomic and alternative splicing changes were systematically analyzed using RNA sequencing (RNA-seq), and selected key genes were validated by RT-qPCR. Functional enrichment analysis was performed on differentially expressed genes (DEGs) and regulated alternative splicing events (RASEs). Expression changes of homeostatic microglial markers and classical polarization markers were analyzed from the existing RNA-seq data. Comparisons were made with public proinflammatory microglial signature gene sets. PARP12 mRNA knockdown led to substantial transcriptomic remodeling: 276 DEGs (107 upregulated and 169 downregulated) and 287 significant RASEs were identified. Downregulated DEGs were significantly enriched in inflammatory pathways such as NF-κB, TNF, and chemokine signaling; most core immune genes (including Ccl2, Ptgs2, and C3) showed coordinated downregulation, whereas a smaller subset (Ccl5, Cd81, and S1pr1) was upregulated. Homeostatic microglial markers (Tmem119, P2ry12, Cx3cr1, Sall1, etc.) did not differ significantly between shPARP12 and shNC cells. Splicing analysis revealed altered splicing of immune/metabolism-related genes such as Fyb, Tnfaip8, Lrch4, and Pkm, but changes in protein isoform ratios were not validated. The overall proinflammatory signature gene set was downregulated after PARP12 knockdown. PARP12 mRNA knockdown is closely associated with attenuation of the proinflammatory transcriptional program and widespread alternative splicing changes in BV2 microglial cells. However, given the unsuccessful knockdown at the protein level, the observed changes cannot be directly attributed to loss of PARP12 protein function. Further mechanistic studies combining CRISPR/Cas9 gene knockout and in vivo pathological models are warranted.
(© 2026. The Author(s).)
Competing Interests: Declarations. Conflict of interest: The authors declare no competing interests.
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Contributed Indexing: Keywords: Alternative splicing; BV2 microglia; PARP12; Proinflammatory responses; RNA sequencing
Substance Nomenclature: 0 (RNA, Messenger)
EC 2.4.2.30 (Poly(ADP-ribose) Polymerases)
0 (Inflammation Mediators)
Entry Date(s): Date Created: 20260814 Date Completed: 20260814 Latest Revision: 20260817
Update Code: 20260817
PubMed Central ID: PMC13476304
DOI: 10.1007/s12035-026-06125-y
PMID: 42601570
Βάση Δεδομένων: MEDLINE
Περιγραφή
ISSN:1559-1182
DOI:10.1007/s12035-026-06125-y