Madaric Juraj, Valachovicova Martina, Paulis Ludovit, Pribojova Jana, Mateova Renata, Sebekova Katarina, Postulkova Luba, Madaricova Terezia, Bucova Maria, Mistrik Martin, Vulev Ivan
National Institute of Cardiovascular Diseases, Slovak Medical University, Bratislava, Slovakia.
Slovak Medical University, Bratislava, Slovakia.
Stem Cell Res Ther. 2017 Jul 12;8(1):165. doi: 10.1186/s13287-017-0622-2.
Asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase, acts as an inhibitor of angiogenesis and is associated with an increased risk of cardiovascular mortality. Administration of stem cells may affect endogenous mechanisms that regulate ADMA production and metabolism. The aim of the present study was to analyze ADMA concentration and changes in oxidative stress in patients with advanced critical limb ischemia (CLI) after bone marrow-derived mononuclear cell (BM-MNC) therapy.
Fifty patients (age 64 ± 11 years, 44 males, 6 females) with advanced CLI (Rutherford category 5 or 6) not eligible for revascularization were treated by intramuscular (n = 25) or intra-arterial (n = 25) injection of 40 ml BM-MNC concentrate. Patients with limb salvage and improved wound healing after 6 months were considered responders to cell therapy. The concentrations of markers of oxidative stress and angiogenesis were analyzed before, and at 3 and 6 months after BM-MNC delivery.
At 6-month follow-up, four patients died of reasons unrelated to stem cell therapy. Among the survivors, 80% (37/46) showed limb salvage and improved wound healing. At 6 months follow-up, ADMA concentration significantly decreased in patients with limb salvage (1.74 ± 0.66 to 0.90 ± 0.49 μmol/L, p < 0.001), in parallel with decreased tumor necrosis factor (TNF)-α (2.22 ± 0.16 to 1.94 ± 0.38 pg/ml, p < 0.001), and increased reduced glutathione (6.96 ± 3.1 to 8.67 ± 4.2 μmol/L, p = 0.02), superoxide dismutase activity (168 ± 50 to 218 ± 37 U/L, p = 0.002), and coenzyme Q10 concentration (468 ± 182 to 598 ± 283 μg/L, p = 0.02). The number of delivered BM-MNCs significantly correlated with the decrease in ADMA concentration at 3 months (p = 0.004, r = -0.48) and the decrease in TNF-α concentration at 6 months (p = 0.03, r = -0.44) after cell delivery. ADMA or TNF-α improvement did not correlate with the number of applied CD34 cells, C-reactive protein concentration, leukocyte count, or the dose of atorvastatin.
The therapeutic benefit of BM-MNC therapy is associated with reduced ADMA levels and oxidative stress. Regulation of the ADMA-nitric oxide axis and improved antioxidant status may be involved in the beneficial effects of stem cell therapy.
The study was approved and retrospectively registered by ISRCTN registry, ISRCTN16096154 . Registered on 26 July 2016.
不对称二甲基精氨酸（ADMA）是一氧化氮合酶的内源性抑制剂，可作为血管生成的抑制剂，并与心血管疾病死亡风险增加相关。干细胞的应用可能会影响调节ADMA产生和代谢的内源性机制。本研究的目的是分析晚期严重肢体缺血（CLI）患者在接受骨髓来源的单核细胞（BM-MNC）治疗后ADMA浓度及氧化应激的变化。
50例（年龄64±11岁，男性44例，女性6例）不符合血运重建条件的晚期CLI（卢瑟福分级5级或6级）患者，通过肌肉注射（n = 25）或动脉内注射（n = 25）40 ml BM-MNC浓缩液进行治疗。6个月后肢体得以保全且伤口愈合改善的患者被视为细胞治疗的有效应答者。在BM-MNC输注前、输注后3个月和6个月分析氧化应激和血管生成标志物的浓度。
在6个月的随访中，4例患者死于与干细胞治疗无关的原因。在幸存者中，80%（37/46）的患者肢体得以保全且伤口愈合改善。在6个月的随访中，肢体得以保全的患者ADMA浓度显著降低（从1.74±0.66降至0.90±0.49 μmol/L，p < 0.001），同时肿瘤坏死因子（TNF）-α降低（从2.22±0.16降至～1.94±0.38 pg/ml，p < 0.001），还原型谷胱甘肽增加（从6.96±3.1升至8.67±4.2 μmol/L，p = 0.02），超氧化物歧化酶活性增加（从168±50升至218±37 U/L，p = 0.002），辅酶Q10浓度增加（从468±182升至598±283 μg/L，p = 0.02）。输注的BM-MNC数量与细胞输注后3个月ADMA浓度的降低（p = 0.004，r = -0.48）以及6个月TNF-α浓度的降低（p = 0.03，r = -0.44）显著相关。ADMA或TNF-α的改善与应用的CD34细胞数量、C反应蛋白浓度、白细胞计数或阿托伐他汀剂量无关。
BM-MNC治疗的益处与ADMA水平降低和氧化应激减轻有关。ADMA-一氧化氮轴的调节及抗氧化状态的改善可能参与了干细胞治疗的有益作用。
本研究已获批准，并在ISRCTN注册中心进行了回顾性注册，注册号为ISRCTN16096154。于2016年7月26日注册。
