未经证实:该研究的目的是通过细胞外囊泡(EV)microRNAs(miRs)揭示细胞相互作用的存在,抑制人角膜内皮(HCE)组织退化的恶性循环。
未经批准:预期,比较,观察性研究。
未经证实:新生儿来源的角膜组织中的miR水平,在大泡性角膜移植术和白内障患者的房水(AqH)中,以及培养的人角膜内皮细胞(hCEC)的培养上清液(CS)和EV,使用3D-Gene人类miR芯片确定,然后使用实时聚合酶链反应进行验证。在细胞miR-34a强制下调后,细胞外释放的miR被分析,通过miR-34a抑制剂或暴露于H2O2。评估衰老相关的分泌表型和线粒体膜电位(MMP)以确定释放的miR的功能特征。
未经授权:减弱HCE变性的功能性miRs的鉴定。
UNASSIGNED:将AqH中的miRs分为2组:1组的表达在新生儿来源的组织中明显降低,而另一组几乎保持不变,独立于衰老。miR-34a和-29家族在前一组中是典型的,而miR-184和-24-3p在后者中是典型的。此外,与以前的miRs相比,在AqH中检测到更多的后者miRs。hCEC中miR-184和-24-3p的丰度也更高,EV,和CS在完全成熟的CD44-/沉闷的hCEC,导致足够的临床组织再生能力在细胞注射治疗。细胞miR-34a的抑制,由于miR-34a抑制剂或暴露于氧化应激,出乎意料地导致miR-184和-24-3p的释放增加。VEGF的分泌,白细胞介素6、单核细胞趋化蛋白-1和MMP在成熟CD44-/暗淡和变性CD44+++hCEC中均被抑制,用miR-184模拟物转染。
UASSIGNED:miR-184向AqH的升高释放可能构成细胞相互作用,防止氧化应激诱导的HCE变性加重,从而维持HCE中的组织稳态。
UNASSIGNED: The objective of the study was to reveal the presence of cellular interplay through extracellular vesicle (EV) microRNAs (miRs), to dampen the vicious cycle to degenerate human corneal endothelium (HCE) tissues.
UNASSIGNED: Prospective, comparative, observational study.
UNASSIGNED: The miR levels in neonate-derived corneal tissues, in the aqueous humor (AqH) of bullous keratoplasty and cataract patients, as well as in the culture supernatant (CS) and EV of cultured human corneal endothelial cells (hCECs), were determined using 3D-Gene human miR chips and then validated using the real-time polymerase chain reaction. The extracellularly released miRs were profiled after the forced downregulation of cellular miR-34a, either by an miR-34a inhibitor or exposure to H2O2. The senescence-associated secretory phenotypes and mitochondrial membrane potential (MMP) were assessed to determine the functional features of the released miRs.
UNASSIGNED: Identification of functional miRs attenuating HCE degeneration.
UNASSIGNED: The miRs in AqH were classified into 2 groups: expression in 1 group was significantly reduced in neonate-derived tissues, whereas that in the other group remained almost constant, independent of aging. The miR-34a and -29 families were typical in the former group, whereas miR-184 and -24-3p were typical in the latter. Additionally, a larger amount of the latter miRs was detected in AqH compared with those of the former miRs. There was also a greater abundance of miR-184 and -24-3p in hCECs, EV, and CS in fully mature CD44-/dull hCEC, leading to sufficient clinical tissue regenerative capacity in cell injection therapy. The repression of cellular miR-34a, either due to miR-34a inhibitors or exposure to oxidative stress, unexpectedly resulted in the elevated release of miR-184 and -24-3p. Secretions of VEGF, interleukin 6, monocyte chemotactic protein-1, and MMP were all repressed in both mature CD44-/dull and degenerated CD44+++ hCEC, transfected with an miR-184 mimic.
UNASSIGNED: The elevated release of miR-184 into AqH may constitute cellular interplay that prevents the aggravation of HCE degeneration induced by oxidative stress, thereby sustaining tissue homeostasis in HCE.