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  • Inhibition of acid‑sensing ion channel 1a attenuates acid‑induced activation of autophagy via a calcium signaling pathway in articular chondrocytes.

Inhibition of acid‑sensing ion channel 1a attenuates acid‑induced activation of autophagy via a calcium signaling pathway in articular chondrocytes.

International journal of molecular medicine (2019-02-06)
Wen-Fan Gao, Ya-Yun Xu, Jin-Fang Ge, Fei-Hu Chen
ABSTRACT

Acid‑sensing ion channel 1a (ASIC1a), member of the degenerin/epithelial sodium channel protein superfamily, serves a critical role in various physiological and pathological processes. The aim of the present study was to examine the role of ASIC1a in the autophagy of rat articular chondrocytes. Autophagy was induced by acidic stimulation in rat articular chondrocytes and the extent of autophagy was evaluated via the expression levels of microtubule‑associated protein 1 light chain 3II, Beclin1 and uncoordinated‑51 like kinase1. Suppression of ASIC1a was achieved using small interfering RNA technology and/or inhibitor psalmotoxin‑1. The expression levels of autophagy markers were measured by western blot analysis and reverse transcription‑quantitative polymerase chain reaction methods. Intracellular calcium ([Ca2+]i) was analyzed using a Ca2+‑imaging method. Additionally, protein expression levels of the Ca2+/calmodulin‑dependent protein kinase kinase β (CaMKKβ)/5'‑monophosphate‑activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway were measured by western blot analysis. The results showed that autophagy was increased in a pH‑ and time‑dependent manner with exposure to an acidic environment. In addition, silencing ASIC1a significantly decreased the expression levels of autophagy makers, accompanied by abrogation of the acid‑induced [Ca2+]i increase. Furthermore, silencing of ASIC1a downregulated the levels of CaMKKβ/β‑actin and phosphorylated (p‑) AMPK/AMPK, and upregulated the levels of p‑mTOR/mTOR. These results indicated that ASIC1a is a potent regulator of autophagy in chondrocytes, which may be associated with decreased Ca2+ influx and the CaMKKβ/AMPK/mTOR pathway.

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Anti-ACCN2, affinity isolated antibody