初发和复发翼状胬肉成纤维细胞分泌基质金属蛋白酶的比较及汉防己甲素对其的影响

Comparison of matrix metalloproteinase secreted by primary and recurrent pterygium fibroblasts and inhibitory effect of tetrandrine

目的检测培养的初、复发翼状胬肉成纤维细胞分泌基质金属蛋白酶(matrix metalloproteinases,MMPs)的情况,并作为观察指标了解汉防己甲素(tetrandrine,Tet)对其的抑制作用。方法相同的条件下培养初、复发翼状胬肉与正常球结膜成纤维细胞,酶联免疫吸附试验检测培养的细胞上清液中MMP-1、MMP-3、MMP-9的含量,同时应用10-5mol·L-1Tet作用于培养的初、复发翼状胬肉成纤维细胞,48h后分析培养液中上述MMPs含量的变化。结果培养的初、复发翼状胬肉成纤维细胞上清中MMP-1含量均较正常球结膜高(均为P<0.05),加入10-5mol·L-1Tet后均明显下降(均为P<0.05),但初发翼状胬肉上清中MMP-1含量仍高于正常球结膜(P<0.05),而复发翼状胬肉上清中MMP-1含量与正常球结膜相比差异无统计学意义(P>0.05);初发翼状胬肉上清中MMP-1含量明显高于复发翼状胬肉,加入Tet后仍较复发翼状胬肉高(均为P<0.05)。培养的初发翼状胬肉成纤维细胞上清中MMP-3含量较正常球结膜高(P<0.05),加入10-5mol·L-1Tet后明显下降(P<0.05),但仍高于正常球结膜(P<0.05);复发翼状胬肉上清中MMP-3含量低于正常球结膜(P<0.05)。培养的初、复发翼状胬肉和正常球结膜成纤维细胞上清中均未检测到MMP-9。结论初、复发翼状胬肉成纤维细胞MMP-1、MMP-3的分泌有明显的差异,Tet对初、复发翼状胬肉成纤维细胞MMP-1及初发翼状胬肉MMP-3的分泌有明显的抑制作用。

Objective To examine the level of matrix metalloproteinase secreted by cultured human primary and recurrent pterygium fibroblasts（HPF）,and investigate the inhibitory effect of tetrandrine（Tet） on its.Methods Human conjunctival fibroblasts（HCF） and HPF were cultured under the same conditions.The levels of MMP-1,MMP-3 and MMP-9 in culture media were determined by enzyme-linked immune sorbent assay with or without the treatment of 10-5 mol·L-1 Tet for 48 hours.Results The protein levels of MMP-1 in primary and recurrent HPF were higher than that in HCF,there were statistical differences（all P0.05）.The amount of MMP-1 was significantly decreased by addition of 10-5 mol·L-1 Tet for 48 hours,there were statistical differences（all P0.05）,but that in primary HPF was higher than that in normal HCF,there was statistical difference（P0.05）,and no difference between recurrent HPF and normal HCF（P0.05）.The protein levels of MMP-1 in primary HPF was higher than that in recurrent HPF,after added the 10-5 mol·L-1 Tet for 48 hours,the former was still higher than the latter,there was statistical difference（P0.05）.The protein levels of MMP-3 in cultured primary HPF were higher than that in normal HCF,there was statistical difference（P0.05）,significantly decreased by addition of 10-5 mol·L-1 Tet for 48 hours,there was statistical difference（P0.05）,but still higher than that of normal HCF,there was statistical difference（P0.05）.The protein levels of MMP-3 in cultured recurrent HPF were lower than that of normal HCF,there was statistical difference（P0.05）.MMP-9 protein was not found in primary and recurrent HPF and HCF conditioned medium.Conclusion The levels of MMP-1 and MMP-3 secreted by primary HPF are different from recurrent HPF.Tet dramatically decrease the MMP-1 level secreted by cultured primary and recurrent HPF and MMP-3 level secreted by primary HPF.