Objective To investigate the effect of hypoxia on acid sensitive ion channel 3 (ASIC3) in nucleus pulposus microenvironment and its mechanism. Methods Primary nucleus pulpocytes were isolated and identified by immunohistochemistry, hematoxylin and eosin (HE) and toluidine blue. Immunohistochemistry, HE, toluidine blue and real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) were used to explore the appropriate hypoxic concentration, apoptosis and FQ-PCR to explore the appropriate action time. The interference and overexpression of ASIC3 vector were constructed, and their efficiency was verified by FQ-PCR and Western blotting. The experiment was divided into the normal group (Control). Hypoxic treatment group (2%O2); shRNA-NC+ 2%O2); shRNA-ASIC3+ 2%O2; Expression of no-load + hypoxic treatment (Vector+ 2%O2); ASIC3 expression + hypoxic treatment group (ASIC3+ 2%O2). The proliferation was detected by cell counting kit-8 (CCK-8), the cell cycle and apoptosis were detected by flow cytometry, the formation of autophagosomes in each group was detected by transmission electron microscopy, the expression of hypoxia inducible factor (HIF)-1 and ASIC3 was detected by immunofluorescence, the expression of HIF-1, light chain 3 (LC3), extracellular signal-regulated kinase 1/2 (ERK1/2) and mitogen-activated protein kinase (MAPK) and phosphorylation were detected by FQ-PCR and Western blotting, and the effect of ASIC3 on HIF-1 and ERK1/2 was detected by immunoco-precipitation. Results Nucleus pulposus cells were successfully isolated with appropriate hypoxic concentration of 2% and appropriate action time of 48 h. Both interference and overexpression vector construction were successful and effective. Compared with the control group, cell proliferation was inhibited in the hypoxic group and ASIC3 expression + hypoxic treatment group, apoptosis was increased, cell cycle arrest was in G1 phase, calcium ion concentration was increased, and HIF-1, LC3, p-ERK1/2 and p-mapk expression were increased, the difference was statistically significant (P<0.05). Conclusion Interference with ASIC3 can inhibit the activation of mitogen-activated protein kinase (MAPK) signal to improve the microenvironment of hypoxic nucleus pulposus cells, which may be one of the pathogenesis of discogenic low back pain. Key words: Nucleus pulposus cells; Discogenic low back pain; Acid-sensing ion channel