【摘要】：研究目的 研究DQ12對(duì)佛波酯（PMA）誘導(dǎo)分化的THP-1細(xì)胞的基質(zhì)金屬蛋白酶-9mRNA表達(dá)、酶活性的影響以及松弛素（relaxin）的干預(yù)作用。 研究方法 RPMI-1640培養(yǎng)基培養(yǎng)THP-1細(xì)胞，于對(duì)數(shù)生長(zhǎng)期接種于6孔板，調(diào)整細(xì)胞濃度8×105/ml。10ng/ml的PMA刺激THP-1細(xì)胞24h后無(wú)血清培養(yǎng)3h，然后分為對(duì)照組、DQ12組、DQ12+R組；對(duì)照組無(wú)血清培養(yǎng)基培養(yǎng)、DQ12組含100ug/mlDQ12的無(wú)血清培養(yǎng)基培養(yǎng)、DQ12+R組含100ug/ml DQ12和10ng/ml的Relaxin培養(yǎng)，，于1h、3h、12h、24h吸取培養(yǎng)液、收獲細(xì)胞。培養(yǎng)液分裝、-80℃凍存，明膠酶譜法檢測(cè)基質(zhì)金屬蛋白酶-9活性；提取THP-1細(xì)胞RNA，qRT-PCR檢測(cè)基質(zhì)金屬蛋白酶-9mRNA的表達(dá)。 研究結(jié)果 DQ12組THP-1細(xì)胞基質(zhì)金屬蛋白酶-9mRNA的表達(dá)于12h開始增加，24h維持在增高水平(P0.01)；與對(duì)照組比較，DQ12組THP-1細(xì)胞基質(zhì)金屬蛋白酶-9活性于1、3、12、24h均增強(qiáng)(P0.01)。 DQ12+R組與DQ12組比較，降低3、12、24基質(zhì)金屬蛋白酶-9mRNA的表達(dá)(P0.05)；基質(zhì)金屬蛋白酶-9活性差異沒有統(tǒng)計(jì)學(xué)意義（p0.05）。 研究結(jié)論 DQ12增加PMA誘導(dǎo)分化的THP-1細(xì)胞的基質(zhì)金屬蛋白酶-9mRNA表達(dá)和酶活性。Relaxin對(duì)DQ12誘導(dǎo)的基質(zhì)金屬蛋白酶-9mRNA表達(dá)增高有抑制作用，對(duì)基質(zhì)金屬蛋白酶-9活性的影響不明顯。本研究結(jié)果提示relaxin可能通過影響肺泡巨噬細(xì)胞基質(zhì)金屬蛋白酶-9表達(dá)干預(yù)矽塵致肺纖維化作用。
[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.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R114

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