Department of Medicine, Royal College of Surgeons in Ireland, Dublin, Ireland.
Department of Medicine, Faculty of Medicine, UKM Medical Centre, Jalan Yaakob Latiff, Bandar Tun Abdul Razak, 56000, Kuala Lumpur, Malaysia.
Sci Rep. 2017 Oct 23;7(1):13803. doi: 10.1038/s41598-017-14310-2.
Alpha-1 antitrypsin (AAT) augmentation therapy involves infusion of plasma-purified AAT to AAT deficient individuals. Whether treatment affects microRNA expression has not been investigated. This study's objectives were to evaluate the effect of AAT augmentation therapy on altered miRNA expression in monocytes and investigate the mechanism. Monocytes were isolated from non-AAT deficient (MM) and AAT deficient (ZZ) individuals, and ZZs receiving AAT. mRNA (qRT-PCR, microarray), miRNA (miRNA profiling, qRT-PCR), and protein (western blotting) analyses were performed. Twenty one miRNAs were differentially expressed 3-fold between ZZs and MMs. miRNA validation studies demonstrated that in ZZ monocytes receiving AAT levels of miR-199a-5p, miR-598 and miR-320a, which are predicted to be regulated by NFκB, were restored to levels similar to MMs. Validated targets co-regulated by these miRNAs were reciprocally increased in ZZs receiving AAT in vivo and in vitro. Expression of these miRNAs could be increased in ZZ monocytes treated ex vivo with an NFκB agonist and decreased by NFκB inhibition. p50 and p65 mRNA and protein were significantly lower in ZZs receiving AAT than untreated ZZs. AAT augmentation therapy inhibits NFκB and decreases miR-199a-5p, miR-598 and miR-320a in ZZ monocytes. These NFκB-inhibitory properties may contribute to the anti-inflammatory effects of AAT augmentation therapy.
α-1 抗胰蛋白酶（AAT）增强疗法包括将血浆纯化的 AAT 输注到 AAT 缺乏的个体中。治疗是否会影响 microRNA 的表达尚未得到研究。本研究的目的是评估 AAT 增强疗法对单核细胞中改变的 microRNA 表达的影响，并探讨其机制。从非 AAT 缺乏（MM）和 AAT 缺乏（ZZ）个体以及接受 AAT 的 ZZ 中分离单核细胞，并进行 mRNA（qRT-PCR、微阵列）、miRNA（miRNA 分析、qRT-PCR）和蛋白质（western blot）分析。在 ZZ 和 MM 之间有 21 个 miRNA 差异表达 3 倍。miRNA 验证研究表明，在接受 AAT 的 ZZ 单核细胞中，miR-199a-5p、miR-598 和 miR-320a 的水平被恢复到与 MM 相似的水平，这些 miRNA 被预测受 NFκB 调节。这些 miRNA 共同调节的靶标在接受体内和体外 AAT 的 ZZ 中呈反式增加。用 NFκB 激动剂体外处理 ZZ 单核细胞可增加这些 miRNA 的表达，并通过 NFκB 抑制降低其表达。与未接受治疗的 ZZ 相比，接受 AAT 的 ZZ 中的 p50 和 p65 mRNA 和蛋白显著降低。AAT 增强疗法抑制 NFκB 并降低 ZZ 单核细胞中的 miR-199a-5p、miR-598 和 miR-320a。这些 NFκB 抑制特性可能有助于 AAT 增强疗法的抗炎作用。
