Cacciolatti Cristina, Meyer-Ficca Mirella L, Southwood Louise L, Meyer Ralph G, Bertolotti Luigi, Zarucco Laura
Am J Vet Res. 2019 Jul;80(7):663-669. doi: 10.2460/ajvr.80.7.663.
To evaluate effects of poly(ADP-ribose) polymerase-1 (PARP1) inhibitors on the production of tumor necrosis factor-α (TNF-α) by interferon-γ (IFN-γ)- and lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (PBMCs) of horses as an in vitro model of inflammation in horses.
1,440 samples of PBMCs from 6 healthy research horses.
From heparinized whole blood samples, PBMC cultures were obtained. An initial dose-response trial on 48 PBMC samples from 2 horses (24 samples each) was used to determine concentrations of IFN-γ and LPS for use as low- and high-level stimulation concentrations. Seventy-two PBMC samples from 6 horses were assigned equally to 1 of 4 PARP1 inhibition categories: no PARP1 inhibitor (PARP1 inhibition control); 2-(()-2-methylpyrrolidin-2-yl)-1-benzimidazole-4-carbozamide dihydrochloride (ABT888);4-(3-(1-(cyclopropanecarbonyl)piperazine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2)-one (AZD2281); or -(6-oxo-5,6-dihydrophenanthridin-2-yl) --dimethylacetamide hydrochloride (PJ34). Samples of PBMCs from each horse and each PARP1 inhibition category were then assigned to 1 of 3 levels of IFN-γ and LPS stimulation: none (control), low stimulation, or high stimulation. After a 24-hour incubation period, a TNF-α ELISA was used to measure TNF-α concentration in the supernatant. Results were compared across treatments and for each horse. Data were analyzed with repeated-measures ANOVA.
Median TNF-α concentration was significantly lower for PJ34-treated, high-level stimulated PBMCs than for PARP1 inhibition control, high-level stimulated PBMCs; however, no other meaningful differences in TNF-α concentration were detected among the inhibition and stimulation combinations.
Findings suggested that PJ34 PARP1 inhibition may reduce TNF-α production in horses, a potential benefit in reducing inflammation and endotoxin-induced damage in horses.
以马的炎症体外模型，评估聚（ADP - 核糖）聚合酶 - 1（PARP1）抑制剂对干扰素 - γ（IFN - γ）和脂多糖（LPS）刺激的马外周血单个核细胞（PBMCs）产生肿瘤坏死因子 - α（TNF - α）的影响。
来自6匹健康研究用马的1440份PBMC样本。
从肝素化全血样本中获取PBMC培养物。对来自2匹马的48份PBMC样本（每匹马24份）进行初始剂量反应试验，以确定用作低水平和高水平刺激浓度的IFN - γ和LPS浓度。将来自6匹马的72份PBMC样本平均分配到4种PARP1抑制类别中的1种：无PARP1抑制剂（PARP1抑制对照）；2 - （（（ - ） - 2 - 甲基吡咯烷 - 2 - 基） - 1 - 苯并咪唑 - 4 - 甲酰胺二盐酸盐（ABT888）；4 - （3 - （1 - （环丙烷羰基）哌嗪 - 4 - 羰基） - 4 - 氟苄基）酞嗪 - 1（2） - 酮（AZD2281）；或 - （6 - 氧代 - 5,6 - 二氢菲啶 - 2 - 基） - N，N - 二甲基乙酰胺盐酸盐（PJ34）。然后将来自每匹马和每种PARP1抑制类别的PBMC样本分配到3种IFN - γ和LPS刺激水平中的1种：无（对照）、低刺激或高刺激。孵育24小时后，使用TNF - α ELISA测量上清液中TNF - α的浓度。对各处理组以及每匹马的结果进行比较。数据采用重复测量方差分析。
PJ34处理的高水平刺激PBMCs的TNF - α浓度中位数显著低于PARP1抑制对照的高水平刺激PBMCs；然而，在抑制和刺激组合之间未检测到其他有意义的TNF - α浓度差异。
研究结果表明，PJ34抑制PARP1可能会降低马体内TNF - α的产生，这在减轻马的炎症和内毒素诱导的损伤方面可能具有潜在益处。
