Effects of butyrate on matrix metalloproteinase-3 expression in intestinal epithelial cell line No.6

Abstract

Objective To investigate the effects of butyrate on the expression of matrix metalloproteinase-3 (MMP-3) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in intestinal epithelial cell line No. 6 (IEC-6) cells. Methods IEC-6 cells of logarithmic growth phase were divided into three groups according to different concentrations of sodium butyrate being used. Low-concentration group, the final concentration of sodium butyrate in medium was 2 mmol/L; high-concentration group, the final concentration of sodium butyrate in medium was 14 mmol/L and control group with no sodium butyrate in medium. After cultured for 24 hours, expression of MMP-3 and TIMP-1 mRNA in IEC-6 cells was measured by reverse transcription-polymerase chain reaction. After cultured for 48 hours, Western blot was employed to examine the expression of MMP-3 and TIMP-1 protein in IEC-6 cells. The differences between groups were compared by ANOVA. Results Expression level of MMP-3 mRNA in group treated with butyrate in concentration of 14 mmol/L was 0. 566±0. 140,inactive form protein of MMP-3 in the group was 0. 756±0. 128, and expression level of active form protein was 0. 255±0. 057, which were higher than those of control group (0. 166±0. 041, 0. 312± 0. 057, 0. 042 ± 0. 038) (P＜0.01, respectively). While, there were no significant differences of those indexes between the control group and low-concentration group. In the high and low-concentration and control group, the levels of TIMP-1 mRNA were 0. 427±0. 042, 0. 383±0. 043 and 0. 355±0. 048,the expression of TIMP-1 protein were 0. 576±0. 140, 0. 524±0. 123 and 0. 355±0. 062, there were no singnificant difference among the three groups (F= 1.97 and 3.10, P＞ 0.05). Conclusions High concentration sodium butyrate could induce the expression of MMP-3 in IEC-6 cells, while not at low concentration. Key words: Enterocolitis, necrotizing; Cell line; Matrix metalloproteinase 3; Butyrates