Department of Clinical Sciences, Cummings School of Veterinary Medicine at Tufts University, North Grafton, Massachusetts, USA.
Michigan State University Veterinary Medical Center, East Lansing, Michigan, USA.
Vet Ophthalmol. 2024 May;27(3):238-247. doi: 10.1111/vop.13143. Epub 2023 Sep 1.
The objective of the study was to compare the ability of aqueous humor (AH) from dogs with primary angle-closure glaucoma (CPACG), companion dogs without overt evidence of CPACG, and Beagles with and without ADAMTS10 open-angle glaucoma (ADAMTS10-OAG) to catalyze or inhibit collagenolysis.
Seventeen normal pet dogs, 27 dogs with CPACG, 19 Beagles with ADAMTS10-OAG, and 4 unaffected Beagles.
A fluorescein-based substrate degradation assay was used to assess AH proteolytic capacity. Samples were then assayed using the same substrate degradation assay, with recombinant activated matrix metalloproteinase-2 (MMP-2) added to measure protease inhibition effects.
For the protease activity assay, relative fluorescence (RF) for AH from normal pet dogs was 13.28 ± 2.25% of control collagenase while RF for AH from dogs with CPACG was 17.47 ± 4.67%; RF was 8.57 ± 1.72% for ADAMTS10-OAG Beagles and 7.99 ± 1.15% for unaffected Beagles. For the MMP-2 inhibition assay, RF for AH from normal dogs was 34.96 ± 15.04% compared to MMP-2 controls, while RF from dogs with CPACG was 16.69 ± 7.95%; RF was 85.85 ± 13.23% for Beagles with ADAMTS10-OAG and 94.51 ± 8.36% for unaffected Beagles. Significant differences were found between dogs with CPACG and both normal pet dogs and dogs with ADAMTS10-OAG and between normal pet dogs and both groups of Beagles.
AH from dogs with CPACG is significantly more able to catalyze proteolysis and inhibit MMP-2 than AH from normal dogs or dogs with ADAMTS10-OAG. Results suggest that pathogenesis may differ between CPACG and ADAMTS10-OAG.
本研究旨在比较原发性闭角型青光眼(CPACG)犬、无明显 CPACG 证据的伴侣犬和具有和不具有 ADAMTS10 开角型青光眼(ADAMTS10-OAG)的比格犬的房水(AH)催化或抑制胶原分解的能力。
17 只正常宠物犬、27 只 CPACG 犬、19 只 ADAMTS10-OAG 比格犬和 4 只未受影响的比格犬。
使用基于荧光的底物降解测定法评估 AH 的蛋白水解能力。然后,使用相同的底物降解测定法对样本进行检测,并添加重组激活基质金属蛋白酶-2(MMP-2)以测量蛋白酶抑制效果。
对于蛋白酶活性测定,正常宠物犬 AH 的相对荧光(RF)为对照胶原酶的 13.28±2.25%,CPACG 犬 AH 的 RF 为 17.47±4.67%;ADAMTS10-OAG 比格犬的 RF 为 8.57±1.72%,未受影响的比格犬的 RF 为 7.99±1.15%。对于 MMP-2 抑制测定,正常犬 AH 的 RF 为 34.96±15.04%,与 MMP-2 对照相比,CPACG 犬的 RF 为 16.69±7.95%;ADAMTS10-OAG 比格犬的 RF 为 85.85±13.23%,未受影响的比格犬的 RF 为 94.51±8.36%。CPACG 犬与正常宠物犬和 ADAMTS10-OAG 犬之间以及正常宠物犬与两组比格犬之间均存在显著差异。
CPACG 犬的 AH 催化蛋白水解和抑制 MMP-2 的能力明显高于正常犬或 ADAMTS10-OAG 犬的 AH。结果表明,CPACG 和 ADAMTS10-OAG 的发病机制可能不同。