Nadeem Muhammad, Kindelin Adam, Mahady Laura, Bhatia Kanchan, Hoda Md Nasrul, Ducruet Andrew F, Ahmad Saif
Department of Neurobiology, Barrow Neurological Institute, SJHMC, Dignity Health, Phoenix, AZ, 85013, USA.
Department of Neurosurgery, Barrow Neurological Institute, SJHMC, Dignity Health, Phoenix, AZ, 85013, USA.
Neuromolecular Med. 2021 Sep;23(3):371-382. doi: 10.1007/s12017-020-08631-1. Epub 2020 Nov 13.
Traumatic optic neuropathy (TON) is characterized by visual dysfunction after indirect or direct injury to the optic nerve following blunt head trauma. TON is associated with increased oxidative stress and inflammation resulting in retinal ganglion cell (RGC) death. Remote ischemic post-conditioning (RIC) has been shown to enhance endogenous protective mechanisms in diverse disease models including stroke, vascular cognitive impairment (VCI), retinal injury and optic nerve injury. However, the protective mechanisms underlying the improvement of retinal function and RGC survival after RIC treatment remain unclear. Here, we hypothesized that RIC therapy may be protective following TON by preventing RGC death, oxidative insult and inflammation in the mouse retina. To carry out the study, mice were divided in three different groups (Control, TON and TON + RIC). We harvested retinal tissue 5 days after TON induction for western blotting and histochemical analysis. We observed increased TON-induced retinal cell death compared with controls by cleaved caspase-3 immunohistochemistry. Furthermore, the TON cohort demonstrated increased TUNEL positive cells which were significantly attenuated by RIC. Immunofluorescence data showed that oxidative stress markers dihydroethidium (DHE), NOX-2 and nitrotyrosine expression were elevated in the TON group relative to controls and RIC therapy significantly reduced the expression level of these markers. Next, we found that the proinflammatory cytokine TNF-α was increased and anti-inflammatory IL-10 was decreased in plasma of TON animals, and RIC therapy reversed this expression level. Interestingly, western blotting of retinal tissue showed that RGC marker Brn3a and tight junction proteins (ZO-1 and Occludin), and AMPKα1 expression were downregulated in the TON group compared to controls. However, RIC significantly increased the expression levels of these proteins. Together these data suggest that RIC therapy activates endogenous protective mechanisms which may attenuate TON-induced oxidative stress and inflammation, and improves BRB integrity.
创伤性视神经病变(TON)的特征是钝性头部外伤后对视神经造成间接或直接损伤后出现视觉功能障碍。TON与氧化应激增加和炎症相关,导致视网膜神经节细胞(RGC)死亡。远程缺血后处理(RIC)已被证明可增强包括中风、血管性认知障碍(VCI)、视网膜损伤和视神经损伤在内的多种疾病模型中的内源性保护机制。然而,RIC治疗后视网膜功能改善和RGC存活的潜在保护机制仍不清楚。在此,我们假设RIC治疗可能通过预防小鼠视网膜中的RGC死亡、氧化损伤和炎症而在TON后具有保护作用。为了开展这项研究,将小鼠分为三个不同的组(对照组、TON组和TON + RIC组)。在TON诱导后5天收集视网膜组织用于蛋白质印迹和组织化学分析。通过裂解的半胱天冬酶-3免疫组织化学,我们观察到与对照组相比,TON诱导的视网膜细胞死亡增加。此外,TON组显示TUNEL阳性细胞增加,而RIC可显著减轻这种情况。免疫荧光数据显示,相对于对照组,TON组中氧化应激标志物二氢乙锭(DHE)、NOX-2和硝基酪氨酸表达升高,而RIC治疗显著降低了这些标志物的表达水平。接下来,我们发现TON动物血浆中促炎细胞因子TNF-α增加,抗炎性IL-10减少,而RIC治疗逆转了这种表达水平。有趣的是,视网膜组织的蛋白质印迹显示,与对照组相比,TON组中RGC标志物Brn3a、紧密连接蛋白(ZO-1和闭合蛋白)以及AMPKα1表达下调。然而,RIC显著增加了这些蛋白质的表达水平。这些数据共同表明,RIC治疗激活了内源性保护机制,这可能减轻TON诱导的氧化应激和炎症,并改善血视网膜屏障的完整性。