In order to explore the role of miR-192 in response to carbonate alkalinity stress in Nile tilapia, real-time PCR, bioinformatics, dual-luciferase reporter assay and miRNA inhibition in vivo were used. The results were as follows:(1) The real-time PCR experiments showed that the expression of miR-192 was significantly down-regulated, while solute carriers16A7,SLC16A7 was significantly up-regulated in gills of Nile tilapia after 6 hours acute alkalinity stress (6 g/L NaHCO₃)(P<0.05); (2) Bioinformatics analysis revealed that SLC16A7 might be the target gene of miR-192; (3) Dual-luciferase reporter assay showed that miR-192 directly regulated SLC16A7 by targeting its 3'UTR; (4) Inhibition of miR-192 significantly increased the mRNA level of SLC16A7 in vivo (P<0.05). In conclusion, miR-192 was involved in the adaptation of Nile tilapia to alkalinity stress,and SLC16A7 was the direct target gene of miR-192. This study provides a basis for exploring the mechanism of miRNAs in the response to alkalinity stress in Nile tilapia.