Raghudeep Eye Hospital, Ahmedabad, Gujarat, India.
Indian J Ophthalmol. 2019 Oct;67(10):1618-1622. doi: 10.4103/ijo.IJO_185_19.
To measure levels of collagen-derived antiangiogenic factors (arresten, canstatin, tumstatin, endostatin) and matrix metalloproteinases (MMP-2 and MMP-9) in anterior lens epithelial cells (LECs) and anterior capsules of children with cataract and persistent fetal vasculature (PFV) as cases and cataract without PFV as controls.
Anterior capsules harboring LECs were collected from pediatric cataract patients with (n = 13) and without PFV (n = 13) during surgery. Samples were immediately subjected to RNA extraction and cDNA preparation. Quantitative real time PCR was performed to determine the mRNA levels of antiangiogenic factors and matrix metalloproteinases. GAPDH (Glyceraldehyde 3-Phosphate Dehydrogenase) and β Actin were used as the housekeeping control. The mRNA levels were expressed as a ratio, using the delta-delta method for comparing the relative expression results between controls and cases. The non-parametric Mann-Whitney U test was applied for statistical evaluation. P values < 0.05 were statistically significant.
The relative mRNA levels of arresten, canstatin, tumstatin, endostatin, MMP-2 and MMP-9 in cases were 6.20E-03 ± 0.003, 1.49E-01 ± 0.02, 1.70E-01 ± 0.007, 3.20E-03 ± 0.003, 1.11E-03 ± 0.0009 and 3.72E-04 ± 0.0001. The mRNA levels of arresten was 1.6 times lower (P = 0.01) while mRNA levels of MMP-2, tumstatin and canstatin were 4, 2.5, and 2.3 times higher in cases than in controls. No change was observed in mRNA levels of MMP-9 and endostatin (P = 0.82).
A significant difference in the levels of arresten, canstatin, tumstatin, and MMP-2 was found in LECs with PFV.
测量具有永存原始玻璃体增生症(PFV)的儿童白内障患者的前晶状体上皮细胞(LEC)和前囊胶原衍生抗血管生成因子(arresten、canstatin、tumstatin、endostatin)和基质金属蛋白酶(MMP-2 和 MMP-9)的水平,并与无 PFV 的白内障患者作对照。
手术中从前发性白内障患儿(PFV 组,n=13;非 PFV 组,n=13)的前囊采集 LEC 。立即进行 RNA 提取和 cDNA 制备。采用实时定量 PCR 测定抗血管生成因子和基质金属蛋白酶的 mRNA 水平。GAPDH(甘油醛 3-磷酸脱氢酶)和β肌动蛋白被用作管家控制。使用 delta-delta 方法比较对照组和病例组的相对表达结果,将 mRNA 水平表示为比值。应用非参数 Mann-Whitney U 检验进行统计学评估。P 值<0.05 为统计学显著。
病例组 arresten、canstatin、tumstatin、endostatin、MMP-2 和 MMP-9 的相对 mRNA 水平分别为 6.20E-03±0.003、1.49E-01±0.02、1.70E-01±0.007、3.20E-03±0.003、1.11E-03±0.0009 和 3.72E-04±0.0001。病例组 arresten 的 mRNA 水平低 1.6 倍(P=0.01),而 MMP-2、tumstatin 和 canstatin 的 mRNA 水平分别高 4、2.5 和 2.3 倍。MMP-9 和 endostatin 的 mRNA 水平无变化(P=0.82)。
在具有 PFV 的 LEC 中,arresten、canstatin、tumstatin 和 MMP-2 的水平存在显著差异。