CAJ | 학술논문

目的探讨丁酸钠对大鼠小肠上皮细胞株-6(intestinal epithelial cell line No.6,IEC-6)细胞基质金属蛋白酶-3(matrix metalloproteinase-3,MMP-3)和组织金属蛋白酶抑制物-1(tissue inhibitor of metalloproteinase-1,TIMP-1)表达的影响,为预防新生儿NEC提供理论依据.方法将对数生长期IEC-6细胞分3组进行实验,低浓度丁酸钠组:培养液中加入终浓度为2 mmol/L的丁酸钠;高浓度丁酸钠组:丁酸钠终浓度为14 mmol/L;另设不含丁酸钠的对照组.共培养24 h后获取细胞,采用逆转录聚合酶链反应技术检测MMP-3和TIMP-1 mRNA的水平.培养48 h后,采用Western印迹法检测MMP-3和TIMP-1蛋白的表达水平.组间差异比较采用单因素方差分析及Dunnett-t和Dunnett's T3进行两两检验.结果高浓度丁酸钠组MMP-3 mRNA水平为0.566±0.140,MMP-3酶原水平为0.756±0.128,MMP-3活性形式蛋白水平为0.255±0.057,均较对照组(分别为0.166±0.041、0.312±0.057和0.042±0.038)明显升高,差异均有统计学意义(P均＜0.01).低浓度丁酸钠组MMP-3 mRNA水平(0.213±0.039)、酶原水平(0.407±0.088)和活性形式蛋白水平(0.103±0.050)与对照组差异均无统计学意义(P＞0.05).高、低浓度丁酸钠组和对照组TIMP-1 mRNA为0.427±0.042,0.383±0.043和0.355±0.048;蛋白水平分别为0.576±0.140,0.524±0.123和0.355±0.062,组间差异均无统计学意义(F=1.97和3.10,P均＞0.05).结论高浓度丁酸钠能选择性诱导IEC-6细胞MMP-3的表达,而低浓度丁酸钠对其没有明显影响.
목적 탐토정산납대대서소장상피세포주-6(intestinal epithelial cell line No.6,IEC-6)세포기질금속단백매-3(matrix metalloproteinase-3,MMP-3)화조직금속단백매억제물-1(tissue inhibitor of metalloproteinase-1,TIMP-1)표체적영향,위예방신생인NEC제공이론의거.방법장대수생장기IEC-6세포분3조진행실험,저농도정산납조:배양액중가입종농도위2 mmol/L적정산납;고농도정산납조:정산납종농도위14 mmol/L;령설불함정산납적대조조.공배양24 h후획취세포,채용역전록취합매련반응기술검측MMP-3화TIMP-1 mRNA적수평.배양48 h후,채용Western인적법검측MMP-3화TIMP-1단백적표체수평.조간차이비교채용단인소방차분석급Dunnett-t화Dunnett's T3진행량량검험.결과 고농도정산납조MMP-3 mRNA수평위0.566±0.140,MMP-3매원수평위0.756±0.128,MMP-3활성형식단백수평위0.255±0.057,균교대조조(분별위0.166±0.041、0.312±0.057화0.042±0.038)명현승고,차이균유통계학의의(P균＜0.01).저농도정산납조MMP-3 mRNA수평(0.213±0.039)、매원수평(0.407±0.088)화활성형식단백수평(0.103±0.050)여대조조차이균무통계학의의(P＞0.05).고、저농도정산납조화대조조TIMP-1 mRNA위0.427±0.042,0.383±0.043화0.355±0.048;단백수평분별위0.576±0.140,0.524±0.123화0.355±0.062,조간차이균무통계학의의(F=1.97화3.10,P균＞0.05).결론고농도정산납능선택성유도IEC-6세포MMP-3적표체,이저농도정산납대기몰유명현영향.
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.