Hemmati Sara, Sadeghi Mohammad Amin, Yousefi-Manesh Hasan, Eslamiyeh Mostafa, Vafaei Ali, Foroutani Laleh, Donyadideh Ghazaleh, Dehpour AhmadReza, Rezaei Nima
Molecular Medicine Interest Group (MMIG), Universal Scientific Education and Research Network (USERN), Tehran, Iran.
School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
J Inflamm Res. 2020 Jul 31;13:411-420. doi: 10.2147/JIR.S258991. eCollection 2020.
Reactive oxygen and nitrogen species (ROS and RNS) are involved in pathologic mechanisms underlying demyelination and exacerbation in multiple sclerosis (MS) lesions. P47phox is the most important subunit of an ROS-producing enzyme (NADPH oxidase) which is reportedly upregulated in MS plaques due to the intense activity of infiltrated immune cells and resident microglia. Leukadherin1 is a specific CD11b/CD18 agonist that inhibits signaling and transmigration of inflammatory cells to sites of injury. Based on this mechanism, we evaluated therapeutic effects of leukadherin1 in an animal model of targeted experimental autoimmune encephalomyelitis (EAE) through focal injection of inflammatory cytokines to the spinal cord.
For model induction, Lewis rats were first immunized with 15µg MOG 1-125 emulsion. Twenty days later, animals were subjected to stereotaxic injection of IFNγ and TNFα to the specific spinal area (T8). One day after injection, all animals presented EAE clinical signs, and their behaviors were monitored for eight days through open-field locomotion and grid-walking tests. Leukadherin1-treated animals received daily intraperitoneal injections of 1mg/kg of the drug. The specific spinal tissues were extracted on day 5 in order to measure nitric oxide (NO), malon di-aldehyde (MDA), and TNFα concentrations alongside P47phox real-time PCR analysis. In addition, spinal sections were prepared for immunohistochemical (IHC) observation of infiltrated leukocytes and activated microglia.
Leukadherin1 exhibited promising improvements in EAE clinical scores and behavioral tests. Demyelination, CD45+ leukocyte infiltration, and Iba1+ microglia activation were reduced in spinal tissues of leukadherin1-treated animals. Furthermore, P47phox expression levels, MDA, and NO amounts were decreased in treated animals. However, TNFα concentrations did not differ following treatment.
Based on our results, we suggest that leukadherin1 may be used as a novel therapeutic agent in tackling the clinical challenge of multiple sclerosis, especially during the acute phase of the disease. This effect was possibly mediated through decreased leukocyte infiltration and oxidative stress.
活性氧和氮物种（ROS和RNS）参与了多发性硬化症（MS）病变中脱髓鞘和病情加重的病理机制。P47phox是一种产生活性氧的酶（NADPH氧化酶）的最重要亚基，据报道，由于浸润的免疫细胞和常驻小胶质细胞的强烈活动，其在MS斑块中上调。白细胞粘附素1是一种特异性CD11b/CD18激动剂，可抑制炎症细胞向损伤部位的信号传导和迁移。基于这一机制，我们通过向脊髓局部注射炎性细胞因子，在靶向实验性自身免疫性脑脊髓炎（EAE）动物模型中评估了白细胞粘附素1的治疗效果。
为诱导模型，首先用15μg MOG 1-125乳剂免疫Lewis大鼠。20天后，对动物进行立体定向注射IFNγ和TNFα至特定脊髓区域（T8）。注射后一天，所有动物均出现EAE临床症状，并通过旷场运动和网格行走试验对其行为进行8天监测。接受白细胞粘附素1治疗的动物每天腹腔注射1mg/kg该药物。在第5天提取特定脊髓组织，以测量一氧化氮（NO）、丙二醛（MDA）和TNFα浓度，并进行P47phox实时PCR分析。此外，制备脊髓切片用于免疫组织化学（IHC）观察浸润的白细胞和活化的小胶质细胞。
白细胞粘附素1在EAE临床评分和行为测试中表现出有前景的改善。在接受白细胞粘附素1治疗的动物的脊髓组织中，脱髓鞘、CD45+白细胞浸润和Iba1+小胶质细胞活化减少。此外，治疗动物的P47phox表达水平、MDA和NO含量降低。然而，治疗后TNFα浓度没有差异。
基于我们的结果，我们建议白细胞粘附素1可作为一种新型治疗剂来应对多发性硬化症的临床挑战，尤其是在疾病的急性期。这种作用可能是通过减少白细胞浸润和氧化应激介导的。
