To explore the effect of human chemokine-like factor (CKLF1) C19 polypeptide on the activation of myofibroblasts in oral submucosal fibrosis (OSF) and the effect on the expression of secreted frizzledrelated protein (sFRP).Methods Separate fibroblasts (FB) and myofibroblasts (MFB) from buccal mucosal tissues of healthy volunteers and OSF patients by tissue block culture method. Immunofluorescence staining to detect the expression of α-SMA and F-actin to identify MFB. MFB were randomly divided into control group, 0.001 mg/L C19 group, 0.01 mg/L C19 group, 0.1 mg/L C19 group, the C19 treatment group at each concentration was treated with a culture medium containing 0.001, 0.01, 0.1 mg/L of CKLF1-C19 polypeptide, and the cells were cultured for 48 h, control cells were cultured normally. The CCK-8 method measures the cell proliferation activity at each time point, after collecting the supernatant, determine the content of hydroxyproline, collagen gel contraction test detects cell contraction activity, Transwell chamber experiment to detect cell migration and invasion ability, real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) and Western blot (Western blot) experiments were performed to determine the mRNA and protein expression levels of sFRP1, sFRP2 and sFRP5 in cells.Results The separated FBand MFB were mostly long fusiform. The MFB cell body was larger and more numerous, and α-SMA was positively expressed. Compared with the control group, MFB treated with CKLF1-C19 polypeptide at 0.001, 0.01, and 0.1 mg/L showed a significant decrease in cell proliferation activity after 48 h and 72 h culture (P＜0.05), the content of hydroxyproline in the supernatant decreased (P＜0.05), the shrinkage area ratio of collagen gel increased significantly (P＜0.05), the number of migrating cells and the number of invading cells were significantly reduced (P＜0.05), the mRNA and protein expression levels of sFRP1 and sFRP2 in cells were significantly up-regulated (P＜0.05), while the changes in sFRP5 mRNA and protein expression levels were not statistically significant (P＞0.05).Conclusion CKLF1-C19 polypeptide can inhibit the proliferation, migration and invasion of myofibroblasts in oral submucosal fibrotic tissues, reduced the activation level of cells, and regulate the expression of sFRP1 and sFRP2