中华风湿病学杂志
中華風濕病學雜誌
중화풍습병학잡지
CHINESE JOURNAL OF RHEUMATOLOGY
2011年
3期
183-187,后插1
,共6页
巫世瑶%黄建林%谢宝钊%王明霞%古洁若
巫世瑤%黃建林%謝寶釗%王明霞%古潔若
무세요%황건림%사보쇠%왕명하%고길약
明胶酶A%明胶酶B%抗原,CD147
明膠酶A%明膠酶B%抗原,CD147
명효매A%명효매B%항원,CD147
Gelatinase A%Gelatinase B%Antigens,CD147
目的 观察来氟米特活性代谢产物(A771726)对豆蔻佛波醇乙酯(PMA)诱导的人单核细胞系(THP-1)细胞CD147、基质金属蛋白酶(MMP)-2及MMP-9表达的影响.方法 THP-1细胞悬浮生长于含10%胎牛血清的RPMI-1640培养液中,细胞密度达5×105/ml时用于实验.细胞无血清化处理12 h后,加入PMA和(或)不同剂量的A771726(5、15、45μg/ml),培养24 b,实时荧光定量聚合酶链反应(PCR)检测细胞CD147、MMP-2及MMP-9 mRNA的表达,流式细胞术检测细胞表而CD147的表达,明胶酶谱法检测细胞培养上清液中MMP-2、MMP-9的活性.多个样本均数比较采用单因素方差分析或Kruskal-Wallis秩和检验.结果 细胞经PMA刺激后CD147、MMP-2及MMP-9三者表达量均较与空白组有增高(P＜0.01).A771726干预后,细胞表面CD147平均荧光强度(MFI)有不同程度的下降,对照组MFI为109.5±3.8,A771726干预组(5、15、45μg/ml)MFI分别为73.3±2.5、64.5±2.3、40.9±2.7(P＜0.01);不同浓度A771726干预后MMP-2、MMP-9 mRNA表达量及细胞培养上清液中MMP-2、MMP-9活性与对照组比较均有不同程度的下降(P＜0.01).各干预组CD147 mRNA表达量与对照组比较,未见下降(P＞0.05).结论 来氟米特活性代谢产物A771726可以抑制PMA诱导的THP-1细胞CD147、MMP-2及MMP-9的表达.
目的 觀察來氟米特活性代謝產物(A771726)對豆蔻彿波醇乙酯(PMA)誘導的人單覈細胞繫(THP-1)細胞CD147、基質金屬蛋白酶(MMP)-2及MMP-9錶達的影響.方法 THP-1細胞懸浮生長于含10%胎牛血清的RPMI-1640培養液中,細胞密度達5×105/ml時用于實驗.細胞無血清化處理12 h後,加入PMA和(或)不同劑量的A771726(5、15、45μg/ml),培養24 b,實時熒光定量聚閤酶鏈反應(PCR)檢測細胞CD147、MMP-2及MMP-9 mRNA的錶達,流式細胞術檢測細胞錶而CD147的錶達,明膠酶譜法檢測細胞培養上清液中MMP-2、MMP-9的活性.多箇樣本均數比較採用單因素方差分析或Kruskal-Wallis秩和檢驗.結果 細胞經PMA刺激後CD147、MMP-2及MMP-9三者錶達量均較與空白組有增高(P＜0.01).A771726榦預後,細胞錶麵CD147平均熒光彊度(MFI)有不同程度的下降,對照組MFI為109.5±3.8,A771726榦預組(5、15、45μg/ml)MFI分彆為73.3±2.5、64.5±2.3、40.9±2.7(P＜0.01);不同濃度A771726榦預後MMP-2、MMP-9 mRNA錶達量及細胞培養上清液中MMP-2、MMP-9活性與對照組比較均有不同程度的下降(P＜0.01).各榦預組CD147 mRNA錶達量與對照組比較,未見下降(P＞0.05).結論 來氟米特活性代謝產物A771726可以抑製PMA誘導的THP-1細胞CD147、MMP-2及MMP-9的錶達.
목적 관찰래불미특활성대사산물(A771726)대두구불파순을지(PMA)유도적인단핵세포계(THP-1)세포CD147、기질금속단백매(MMP)-2급MMP-9표체적영향.방법 THP-1세포현부생장우함10%태우혈청적RPMI-1640배양액중,세포밀도체5×105/ml시용우실험.세포무혈청화처리12 h후,가입PMA화(혹)불동제량적A771726(5、15、45μg/ml),배양24 b,실시형광정량취합매련반응(PCR)검측세포CD147、MMP-2급MMP-9 mRNA적표체,류식세포술검측세포표이CD147적표체,명효매보법검측세포배양상청액중MMP-2、MMP-9적활성.다개양본균수비교채용단인소방차분석혹Kruskal-Wallis질화검험.결과 세포경PMA자격후CD147、MMP-2급MMP-9삼자표체량균교여공백조유증고(P＜0.01).A771726간예후,세포표면CD147평균형광강도(MFI)유불동정도적하강,대조조MFI위109.5±3.8,A771726간예조(5、15、45μg/ml)MFI분별위73.3±2.5、64.5±2.3、40.9±2.7(P＜0.01);불동농도A771726간예후MMP-2、MMP-9 mRNA표체량급세포배양상청액중MMP-2、MMP-9활성여대조조비교균유불동정도적하강(P＜0.01).각간예조CD147 mRNA표체량여대조조비교,미견하강(P＞0.05).결론 래불미특활성대사산물A771726가이억제PMA유도적THP-1세포CD147、MMP-2급MMP-9적표체.
Objective To investigate the effects of the leflunomide active metabolite (A771726) on the expression of phorbol-12-myristate-13-acetate (PMA) -induced CD147, matrix metallo-proteinase (MMP)-2 and MMP-9 on THP-1 cells. Methods THP-1 cells were cultured in RPMI-1640 medium supplemented with 10％ fetal bovine serum. For all experiments, THP-1 cells were cultured at an initial density of 5×105/ml. Before A771726 treatment, cells were cultured with serum-free RPMI-1640 medium for 12 h, THP-1cells were co-cultured with PMA at three different concentrations of A771726 (5, 15 , 45 μg/ml) for 24 h.The mRNA expression of CD147, MMP-2 and MMP-9 was measured by real-time PCR. CD147 expression on the cells were evaluated by flow cytometric analysis. The activity of MMP-2 and MMP-9 were evaluated by gelatin zymography. Statistical differences among the groups were tested by one-way ANOVA or KruskalWallis test. Results The expression of CD147, MMP-2 and MMP-9 were upgraded by the PMA. The expression of CD147 on THP-1 cells was inhibited significantly by A771726 in a dose-dependent pattern (P＜0.01). The mean fluorescence intensity (MFI) of CD147 in positive control group was 109.5±3.8, the MFI in A771726 (5, 15, 45 μg/ml) group were 73.3±2.5, 64.5±2.3, 40.9±2.7, respectively. The expression of MMP-2, MMP-9 mRNA and the activity of MMP-2, MMP-9 in the supernatant was inhibited significantly by A771726 (P＜0.01). The expression of CD147 mRNA was not inhibited significantly by A771726 (P＞0.05).Conclusion Leflunomide active metabolite (A771726) can inhibit the expression of PMA-induced CD147,MMP-2 and MMP-9 on THP-1 Cells.
