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.

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