DQ12对THP-1细胞基质金属蛋白酶-9的影响及松弛素干预作用
发布时间：2018-08-13 13:34
【摘要】：研究目的 研究DQ12对佛波酯（PMA）诱导分化的THP-1细胞的基质金属蛋白酶-9mRNA表达、酶活性的影响以及松弛素（relaxin）的干预作用。 研究方法 RPMI-1640培养基培养THP-1细胞，于对数生长期接种于6孔板，调整细胞浓度8×105/ml。10ng/ml的PMA刺激THP-1细胞24h后无血清培养3h，然后分为对照组、DQ12组、DQ12+R组；对照组无血清培养基培养、DQ12组含100ug/mlDQ12的无血清培养基培养、DQ12+R组含100ug/ml DQ12和10ng/ml的Relaxin培养，，于1h、3h、12h、24h吸取培养液、收获细胞。培养液分装、-80℃冻存，明胶酶谱法检测基质金属蛋白酶-9活性；提取THP-1细胞RNA，qRT-PCR检测基质金属蛋白酶-9mRNA的表达。 研究结果 DQ12组THP-1细胞基质金属蛋白酶-9mRNA的表达于12h开始增加，24h维持在增高水平(P0.01)；与对照组比较，DQ12组THP-1细胞基质金属蛋白酶-9活性于1、3、12、24h均增强(P0.01)。 DQ12+R组与DQ12组比较，降低3、12、24基质金属蛋白酶-9mRNA的表达(P0.05)；基质金属蛋白酶-9活性差异没有统计学意义（p0.05）。 研究结论 DQ12增加PMA诱导分化的THP-1细胞的基质金属蛋白酶-9mRNA表达和酶活性。Relaxin对DQ12诱导的基质金属蛋白酶-9mRNA表达增高有抑制作用，对基质金属蛋白酶-9活性的影响不明显。本研究结果提示relaxin可能通过影响肺泡巨噬细胞基质金属蛋白酶-9表达干预矽尘致肺纤维化作用。
[Abstract]:Objective to study the effect of DQ12 on matrix metalloproteinase-9 mRNA expression and enzyme activity in THP-1 cells induced by phorbol ester (PMA) and the effect of relaxin (relaxin) on the expression of matrix metalloproteinase-9 mRNA. Methods THP-1 cells were cultured on RPMI-1640 medium and inoculated on 6-well plate in logarithmic growth period. The THP-1 cells were stimulated with 8 脳 105/ml.10ng/ml PMA for 24 h and cultured without serum for 3 h. Then they were divided into control group (DQ12 group) and DQ12R group (control group). The control group was cultured in serum-free culture medium (DQ12) and the serum-free medium containing 100ug/mlDQ12 (DQ12R group) was cultured with Relaxin and 100ug/ml DQ12, and the cells were harvested at 1h, 3h, 12h, 12h, 24h, respectively. The activity of matrix metalloproteinase-9 was detected by gelatinase spectrum, and the expression of matrix metalloproteinase-9 mRNA was detected by qRT-PCR in THP-1 cells. Results the expression of matrix metalloproteinase-9 mRNA in THP-1 cells in DQ12 group was increased at 12 h and maintained at elevated level at 24 h (P0.01). Compared with the control group, the activity of matrix metalloproteinase-9 in THP-1 cells in DQ12 group was significantly increased at 24 h (P0.01). The expression of matrix metalloproteinase-9 mRNA in DQ12 R group was significantly lower than that in DQ12 group (P0.05), but there was no significant difference in matrix metalloproteinase-9 activity between DQ12 R group and DQ12 group (p0.05). Conclusion DQ12 can increase the expression of matrix metalloproteinase-9 mRNA in differentiated THP-1 cells induced by PMA and inhibit the expression of matrix metalloproteinase-9 mRNA induced by DQ12. The effect on the activity of matrix metalloproteinase-9 was not obvious. These results suggest that relaxin may interfere with pulmonary fibrosis induced by silica by affecting the expression of matrix metalloproteinase-9 in alveolar macrophages.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R114
[Abstract]:Objective to study the effect of DQ12 on matrix metalloproteinase-9 mRNA expression and enzyme activity in THP-1 cells induced by phorbol ester (PMA) and the effect of relaxin (relaxin) on the expression of matrix metalloproteinase-9 mRNA. Methods THP-1 cells were cultured on RPMI-1640 medium and inoculated on 6-well plate in logarithmic growth period. The THP-1 cells were stimulated with 8 脳 105/ml.10ng/ml PMA for 24 h and cultured without serum for 3 h. Then they were divided into control group (DQ12 group) and DQ12R group (control group). The control group was cultured in serum-free culture medium (DQ12) and the serum-free medium containing 100ug/mlDQ12 (DQ12R group) was cultured with Relaxin and 100ug/ml DQ12, and the cells were harvested at 1h, 3h, 12h, 12h, 24h, respectively. The activity of matrix metalloproteinase-9 was detected by gelatinase spectrum, and the expression of matrix metalloproteinase-9 mRNA was detected by qRT-PCR in THP-1 cells. Results the expression of matrix metalloproteinase-9 mRNA in THP-1 cells in DQ12 group was increased at 12 h and maintained at elevated level at 24 h (P0.01). Compared with the control group, the activity of matrix metalloproteinase-9 in THP-1 cells in DQ12 group was significantly increased at 24 h (P0.01). The expression of matrix metalloproteinase-9 mRNA in DQ12 R group was significantly lower than that in DQ12 group (P0.05), but there was no significant difference in matrix metalloproteinase-9 activity between DQ12 R group and DQ12 group (p0.05). Conclusion DQ12 can increase the expression of matrix metalloproteinase-9 mRNA in differentiated THP-1 cells induced by PMA and inhibit the expression of matrix metalloproteinase-9 mRNA induced by DQ12. The effect on the activity of matrix metalloproteinase-9 was not obvious. These results suggest that relaxin may interfere with pulmonary fibrosis induced by silica by affecting the expression of matrix metalloproteinase-9 in alveolar macrophages.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R114
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