重组MMP-2干预后人肾系膜细胞对单核细胞趋化及黏附的影响

Effects of human renal mesangial cells on chemotaxis and adhesion of monocytes after intervened by recombinant MMP-2

目的观察重组基质金属蛋白酶2(MMP-2)干预后人肾系膜细胞(HRMCs)对单核细胞U937趋化及黏附的影响。方法分别用不同浓度(0、0.625、1.25、2.5、5ng/ml)重组MMP-2干预体外培养的HRMCs,6h后行U937趋化及黏附实验。用不同浓度(0、0.5、1、2、3、4、5ng/ml)重组MMP-2干预HRMCs,6h后采用RT-PCR、Western blot检测HRMCs的fractalkine(Fkn)mRNA及其蛋白表达。结果 0、0.625、1.25、2.5、5ng/ml重组MMP-2干预后,每高倍视野的趋化U937细胞数分别为6.55±0.51、14.21±0.95、23.54±1.56、34.48±1.47、48.52±3.55,黏附U937细胞数分别为4.67±0.88、3.33±0.33、2.67±0.56、2.00±0.34、1.33±0.45。Fkn mRNA及蛋白表达水平随MMP-2干预浓度的增加而增加。结论重组MMP-2干预后,HRMCs对U937的趋化作用增强,黏附作用减弱。

Objective To investigate the effects of human renal mesangial cells（HRMCs） on chemotaxis and adhesion of U937 monocytes after intervened by recombinant matrix metalloproteinase-2（MMP-2）. Methods Chemotaxis test and adhesion test of U937 monocytes were performed after HRMCs were intervened by different concentrations（0,0. 625,1.25,2. 5,5 ng/ml） of recombinant MMP-2 in vitro for 6 hours. The expressions of fractalkine（Fkn） mRNA and protein in HRMCs were detected by RT-PCR and Western blot, respectively, after HRMCs were intervened by different concentrations（0,0. 5,1,2,3,4,5 ng/ml） of recombinant MMP-2 in vitro for 6 hours. Results After intervened by different concentrations（0,0. 625,1.25,2. 5,5 ng/ml） of recombinant MMP-2, the counts of chemotactic U937 monocytes in a high power field were 6.55 ~ 0.51,14. 21 ~ 0. 95,23. 54± 1.56,34. 48＋1.47,48. 52±3.55, and those of adherent U937 monocytes were 4. 67±0. 88,3. 33±0. 33,2.67±0. 56,2.00±0. 34,1.33±0. 45, respectively. The expressions of Fkn mRNA and protein in HRMCs were increased along with increasing of recombinant MMP-2 concentration. Conclusion HRMCs can increase chemotaxis and decrease adhesion of U937 monocytes after intervened by recombinant MMP-2.