| 杜航航,张恒术.秋水仙碱对人瘢痕疙瘩成纤维细胞MMP-1､MMP-2表达的影响[J].中国烧伤创疡杂志,2018,(1):48~54. |
| DOI： |
| 中文关键词: 秋水仙碱 瘢痕疙瘩 成纤维细胞 基质金属蛋白酶 影响 实验研究 |
| 英文关键词:Colchicine Keloid Fibroblast MMP Influence |
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| 中文摘要: |
| 【摘要】 目的 研究探讨秋水仙碱对体外培养人瘢痕疙瘩成纤维细胞(keloid fibroblast, KFB) 内基质金属蛋白酶-1(matrix metalloproteinase-1, MMP-1)基质金属蛋白酶-2 (MMP-2) 和基质金属蛋白酶-9 (MMP-9) 表达的影响?方法 取7 例患者的瘢痕疙瘩组织进行体外成纤维细胞培养, 待细胞培养至第3 代时, 加入浓度为0.125%的胰蛋白酶消化液浸泡至细胞质回缩?细胞间隙增大后, 转种于75.0 cm2 的培养瓶中继续培养; 待细胞基本融合成片(密度达85%时) 后, 分别将每例患者瘢痕组织的传代培养细胞分为A 组?B 组?C 组?D 组及E组, A 组加入含胎牛血清的DMEM 培养基继续培养备用, B 组?C 组?D 组?E 组加入含胎牛血清的DMEM 培养基及浓度分别为2?4?8?16 μg/ mL 的秋水仙碱继续培养24 h (37℃, 5.0% CO2 ) 后备用?采用逆转录聚合酶链反应(real-time PCR, RT-PPCR) 法检测基质金属蛋白酶mRNA 的表达情况, Western Blot 法检测基质金属蛋白酶蛋白的表达情况?结果 A 组?B 组?C 组?D 组?E 组KFB 中MMP-1 mRNA 的表达水平分别为(0.174±0.016)?(0.282±0.015)?(0.369±0.022)?(0.541±0.027) 及(0.672±0.020), MMP-2 mRNA 的表达水平分别为(0.201±0.033)?(0.371±0.026)?(0.548±0.031)?(0.653 ±0.019) 及(0.796±0.024), MMP-1 蛋白的表达水平分别为(0.127±0.031)?(0.205±0.013)?(0.293±0.027)?(0.349±0.034) 及(0.512±0.035), MMP-2 蛋白的表达水平分别为(0.278±0.023)?(0.395±0.036)?(0.526±0.025)?(0.673±0.014)及(0.792±0.037), 而MMP-9 mRNA 及MMP-9 蛋白均未见表达?与A 组对比, B 组?C 组?D 组及E 组KFB中MMP-1 mRNA?MMP-2mRNA?MMP-1 蛋白及MMP-2 蛋白的表达水平均较高, P均< 0.01, 差异具有统计学意义, 且随着秋水仙碱浓度的增高, B 组?C 组?D 组及E 组KFB 中MMP-1 mRNA?MMP-2 mRNA?MMP-1蛋白及MMP-2 蛋白的表达水平均呈逐渐增高的趋势, P均< 0.01, 差异具有统计学意义?结论 秋水仙碱可提高KFB 中MMP-1 与MMP-2 的表达水平, 从而达到抗纤维化?抑制瘢痕疙瘩形成的目的, 且在一定浓度范围内, 其作用随着秋水仙碱浓度的增加而逐渐增强, 有望成为临床治疗瘢痕疙瘩的一线药物? |
| 英文摘要: |
| 【Abstract】 Objective To investigate the influence of colchicine on the expression of MMP-1（matrix metalloproteinase-1，MMP-1), MMP-2 and MMP-9 in human KFB(keloid fibroblast) cultured in vitro. Methods Keloid tissues of 7 patients were taken for in vitro fibroblast culture.When cells basically fused into pieces (85% of concentration), passage cells in scar tissues of each patients were divided into 5 groups: A,B,C,D and E. DMEM?culture?media with fetal calf serum was added to Cells in group A to culture them for later use. DMEM?culture?media with fetal calf serum and colchicnie of concentration of 2, 4, 8 and 16μg/ml were added to cells in group B, C, D and E to culture them for 24 hours for later use. RT-PCR (Real-time PCR) was adopted to detect the expression of MMP mRNA and Western blot was adopted to detect the expression of MMP. Results The expression levels of MMP-1 mRNA in KFB in group A, B,C,D and E were （0.174±0.016）,（0.282±0.015）,（0.369±0.022）,（0.541±0.027）and（0.672±0.020) respectively. The expression levels of MMP-2 mRNA in KFB in group A, B,C,D and E were （0.201±0.033）,（0.371±0.026）,（0.548±0.031）,（0.653±0.019）and（0.796±0.024) respectively. The expression levels of MMP-1 in KFB in group A, B,C,D and E were （0.127±0.031）,（0.205±0.013）,（0.293±0.027）,（0.349±0.034）and（0.512±0.035）respectively. The expression levels of MMP-2 in KFB in group A, B,C,D and E were (0.278±0.023）,（0.395±0.036）,（0.526±0025）,（0.673±0.014）and（0.792±0.037）respectively. No expression of MMP-9 mRNA and MMP-9 was observed. The expression levels of MMP-1 mRNA,MMP-2 mRNA, MMP-1 and MMP-2 in KFB in group B, C, D and E were higher than that of group A and the results showed statistically significant difference(P＜0.01). As the concentration of colchicine increased, the expression levels of MMP-1 mRNA,MMP-2 mRNA, MMP-1 and MMP-2 in KFB in group B, C, D and E showed a trend of increase too. The differences were statistically significant(P＜0.01). Conclusion Colchicine can increase the expression level of MMP-1 and MMP-2 in KFB，thus achieving the goal of anti-fibrosis and inhibiting the formation of keloid. In a certain concentration range, the effect of colchicine could be enhanced with the increase of its concentration, so it is expected to be a preferred drug for clinical treatment of keloid. |
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