What type of study is this?

This is a Animal Study study (also classified as: In Vitro Study).

What question did this study set out to answer?

To investigate how TRPC3-mediated Ca2+/NF-κB pathway regulates autophagy in intervertebral disc degeneration (IDD).

May 7, 2026Open Access

TRPC3 induces intervertebral disc degeneration by mediating the Ca2+/NF-κB pathway to inhibit autophagy

Key Points

To investigate how TRPC3-mediated Ca2+/NF-κB pathway regulates autophagy in intervertebral disc degeneration (IDD).
Establishment of IDD rat model using annulus fibrosus puncture method.
Local intraspinal injection of AAV-shRNA targeting TRPC3 in rats.
Transfection of human nucleus pulposus cells with TRPC3 siRNA and treatment with NF-κB inhibitors and autophagy modulators.
Use of micro-computed tomography, immunohistochemistry, and other techniques for analysis.
TRPC3 expression significantly increased in IDD rat models.
TRPC3 knockdown improved histopathological damage and upregulated autophagy markers.
siRNA TRPC3 reduced Ca2+ levels caused by IL-1β and inhibited NF-κB activation.
Increased levels of aggrecan and collagen II with decreased MMP-13 expression following TRPC3 knockdown.

Abstract

BACKGROUND: Intervertebral disc degeneration (IDD) is the primary cause of lower back pain. Transient receptor potential canonical 3 (TRPC3) is a nonselective cation channel permeable to Ca2+. OBJECTIVES: This study explores the mechanisms by which the TRPC3-mediated Ca2+/nuclear factor kappa B (NF-κB) pathway regulates autophagy in IDD. MATERIAL AND METHODS: An IDD rat model was established using the annulus fibrosus puncture method and was treated with local intraspinal injection of adeno-associated virus (AAV)-shRNA targeting TRPC3. Primary human nucleus pulposus cells (NPCs) were transfected with TRPC3 siRNA and subsequently treated with pyrrolidine dithiocarbamate (PDTC; an NF-κB inhibitor), rapamycin (RAPA), or 3-methyladenine (3-MA), respectively. Micro-computed tomography (micro-CT), hematoxylin and eosin (H&E) staining, immunohistochemistry, western blotting, transmission electron microscopy (TEM), and flow cytometry were performed. RESULTS: TRPC3 expression was significantly increased in IDD rats (p < 0.05). TRPC3 shRNA ameliorated histopathological damage in IDD rats and promoted the expression of autophagy-related protein 5 (ATG5), Beclin-1, and LC3-II (all p < 0.05). In vitro, interleukin-1 beta (IL-1β) increased Ca2+ levels, siRNA TRPC3 reduced them, and PDTC further decreased them (p < 0.05). In addition, siRNA TRPC3 increased the expression of ATG5, Beclin-1, and the LC3-II/LC3-I ratio and inhibited phosphorylation of p-NF-κB p65 in NPCs (p < 0.05). Transmission electron microscopy and flow cytometry showed that siRNA TRPC3-induced autophagy promoted apoptosis in NPCs (p < 0.05). Furthermore, siRNA TRPC3 increased the levels of aggrecan and collagen II and decreased matrix metalloproteinase-13 (MMP-13) expression (p < 0.05). CONCLUSIONS: TRPC3 exacerbates IDD by inhibiting protective autophagy via activation of the Ca2+/NF-κB signaling pathway. Knockdown of TRPC3 promotes autophagy, which in turn influences NPC apoptosis and extracellular matrix (ECM) metabolism. This study offers potential novel strategies for IDD prevention and treatment.

Read Full Paperexternally

Ask AI

Helpful

Bookmark

View Full Paper