〔Abstract〕 Objective To investigate whether the expression of Piezo1 in bulbar conjunctival fibroblasts contributes to the development of conjunctivochalasis (CCH). Methods Primary bulbar conjunctival fibroblasts from CCH patients and normal controls (NC) were obtained by tissue block applanation, purified and then subjected to fibroblast identification. Immunohistochemistry was used to detect the expression of Piezo1 in the NC and CCH groups. Immunofluorescence was used to detect the expression of Piezo1 in bulbar conjunctival fibroblasts in the CCH and NC groups, as well as the expression of MMP-9 after the intervention of the Piezo1 agonist, Yoda1. qRT-PCR was used to detect the expression of mRNA of Piezo1 and MMP-9 in the CCH and NC groups, as well as the expression of mRNA of MMP-9 after the intervention of Yoda1. In addition, confocal microscopy was used to detect Ca2+influx in bulbar conjunctival fibroblasts in the NC group after Yoda1 intervention. Membrane clamp was used to detect the current amplitude of NC bulbar conjunctival fibroblasts after Yoda1 intervention to verify the functional expression of Piezo1. Results Primary bulbar conjunctival fibroblasts were successfully cultured. Immunohistochemistry revealed that Piezo1 protein expression was elevated in the CCH group (P<0.01). qRT-PCR assay revealed that mRNA expression of Piezo1 was elevated in the CCH group (P<0.01), mRNA expression of MMP-9 was elevated in the CCH group (P<0.01), and mRNA expression of MMP-9 was elevated in the CCH+Yoda1 group (P<0.01). Immunofluorescence detection of Piezo1 revealed enhanced fluorescence in the CCH group relative to the NC group (P<0.05), and the fluorescence of MMP-9 was enhanced by the addition of Yoda1 stimulation compared with that of the CCH group (P<0.05). Confocal microscopy of NC group bulbar conjunctival fibroblasts after the addition of Yoda1 showed enhanced fluorescence of Piezo1 (P<0.05). The addition of Yoda1 activated enhanced current amplitude, showed by membrane clamp detection of NC group bulbar conjunctival fibroblasts (P<0.000 1). Conclusion Mechanical damage produced by conjunctival tissues in conjunctivochalasis upregulates Piezo1 expression, and Piezo1 activation may amplify the mechanical damage caused by CCH.