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Matrix metalloproteinase-2 and -9 activities in the human lens epithelial cells and serum of steroid induced posterior subcapsular cataracts

机译:类固醇诱导的后囊性白内障在人晶状体上皮细胞和血清中的基质金属蛋白酶2和-9活性

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Purpose: To evaluate the level of matrix metalloproteinase (MMP)-2 and MMP-9 activities in patients with steroid induced posterior subcapsular cataract (PSC). Methods: This prospective, observational study comprised of 156 patients having either steroid induced PSC (n=50) or non-steroidal PSC (n=106) were performed to evaluate the level of MMP-2 and MMP-9 activities in the lens epithelial cells (LECs) and the serum. Anterior lens capsules harboring LECs were obtained during phacoemulsification and peripheral blood was collected from patients before administration of anesthesia. Serum was separated by centrifugation at 10,000× g for 15 min at 4 °C. The LECs and serum samples were processed to analyze MMP-2 and MMP-9 activities using succinylated gelatin assay. Quantitative real time-PCR (qRT–PCR) was performed to determine the mRNA levels of MMP-2 and MMP-9 in LECs. The mRNA levels were expressed as a ratio, using the delta-delta method for comparing the relative expression results between cases with steroid induced PSC and cases with non-steroidal PSC. MMP-2 and MMP-9 levels were also compared in the two groups using immunolocalization. Results: The level of MMP-2 and MMP-9 activity was found to be high in LECs and serum of cases with steroid induced PSC. Further in all steroid induced cases, a 1.4 fold increase was observed in MMP-2 activity in LECs and a 1.4 fold increase in MMP-9 activity in the serum. Both qRT–PCR and immunolocalization showed increased expression of MMP-2 and MMP-9 activity. Conclusions: MMP-2 and MMP-9 activity in both LECs and serum was significantly higher in cases with steroid induced PSC. The possible use of MMP-9 as a non-invasive biomarker in ascertaining the presence of steroid induced PSC should be evaluated using a larger sample size.
机译:目的:评估类固醇性后囊性白内障(PSC)患者的基质金属蛋白酶(MMP)-2和MMP-9活性。方法:这项前瞻性,观察性研究包括156例接受类固醇诱导的PSC(n = 50)或非类固醇PSC(n = 106)的患者,以评估晶状体上皮中MMP-2和MMP-9的水平细胞(LEC)和血清。在超声乳化术中获得了具有LEC的前晶状体囊,并在麻醉前从患者中收集了外周血。通过在4°C下以10,000×g离心15分钟来分离血清。使用琥珀酰明胶测定法处理LEC和血清样品以分析MMP-2和MMP-9活性。进行实时定量PCR(qRT-PCR)以确定LEC中MMP-2和MMP-9的mRNA水平。使用delta-delta方法将mRNA水平以比率表示,以比较类固醇诱导的PSC患者和非类固醇PSC患者之间的相对表达结果。还使用免疫定位比较了两组中的MMP-2和MMP-9水平。结果:在类固醇诱导的PSC患者的LEC和血清中发现MMP-2和MMP-9活性较高。此外,在所有类固醇诱导的病例中,LEC中的MMP-2活性增加了1.4倍,血清中MMP-9活性增加了1.4倍。 qRT-PCR和免疫定位均显示MMP-2和MMP-9活性表达增加。结论:类固醇诱导的PSC患者的LEC和血清中的MMP-2和MMP-9活性均显着较高。应该使用更大的样本量来评估MMP-9作为非侵入性生物标志物在确定类固醇诱导的PSC的存在中的可能用途。

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