The neuroprotective role of 4R-cembranoid during systemic inflammation.

4R-西松烷内酯在全身炎症过程中的神经保护作用。

• 批准号: 10087943
• 负责人: WANDA VELEZ-TORRES
• 金额: $ 8.96万
• 依托单位: UNIVERSIDAD CENTRAL DEL CARIBE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10087943
• 关键词: Acetylcholine; Acute; Aftercare; Animals; Anti-Inflammatory Agents; Antiinflammatory Effect; Behavioral; Binding; Blood - brain barrier anatomy; Brain; Brain region; CCL2 gene; CREB1 gene; Cells; Cerebral Ischemia; Cerebral cortex; Chemosensitization; Cholinergic Receptors; Data; Differentiation Antigens; Endotoxins; Excision; Flow Cytometry; Hippocampus (Brain); Hour; Immune; Immunohistochemistry; In Vitro; Infarction; Infiltration; Inflammation; Inflammatory; Inflammatory Response; Injections; Injury; Ischemic Stroke; Knockout Mice; Label; Lipopolysaccharides; Measures; Mediating; Membrane; Memory; Memory impairment; Modeling; Mus; NF-kappa B; Neurons; Phase; Phosphorylation; Phosphotransferases; Plasma; Production; Property; Public Health; Rattus; Recovery; Research; Rodent Model; Role; Saline; Serum; Signal Transduction; Signaling Protein; Slice; TNF gene; Testing; Therapeutic; Therapeutic Agents; Time; Tissues; Traumatic Brain Injury; Vagus nerve structure; cell injury; cytokine; design; extracellular; improved; in vivo; intraperitoneal; macrophage; monocyte; neuroinflammation; neuronal survival; neuroprotection; neurotoxicity; novel; object recognition; receptor; repaired; systemic inflammatory response; treatment group

Neuroinflammation is an important feature of cerebral ischemia. However, persistent inflammation can contribute to progressive tissue damage during the recovery phase. The bacterial lipopolysaccharide endotoxin (LPS) has been widely used as a model of systemic inflammation in mice. Intraperitoneal LPS injection to animals has been shown to produce an inflammatory signal in brain resulting in an increased production of pro-inflammatory cytokines and memory deficit. Activation of the a7 acetylcholine receptors (a7) has been shown to reduce the release of pro-inflammatory cytokines in vivo. Serum TNF-α levels were significantly higher in a7 knockout mice (a7-K0) compared to WT two hours after LPS endotoxin administration. In a traumatic brain injury model, the a7-K0 mice had significantly higher plasma levels of TNF-a, IL- 1b than WT mice at 24hr post injury. Our research group has identified a natural cembranoid, 4R, with neuroprotective properties. 4R protects against neuronal toxicity in vivo and significantly decreases infarction size in a rodent model of ischemic stroke. Our preliminary data shows that 4R treatment significantly lower serum TNF-α, IL-1b, and brain MCP-1 levels in mice after LPS-induced inflammation. Our hypothesis states that 4R treatment decreases LPS- induced neuroinflammation and improves memory function in mice by potentiation of the a7 receptor. The following specific aims are designed to test this hypothesis: 1) To investigate if 4R decreases neuroinflammation in mice and whether the effect is mediated by the a7 receptor. The effect of 4R will be examined by injecting LPS followed by 1) saline, 2) vehicle, or 3) 4R. Pro-and anti-inflammatory cytokines as well as phosphorylation of Akt, NF-kB, and CREB signaling proteins will be measured in cerebral cortex and hippocampus homogenates. 2) To examine the role of 4R and the a7 receptor in monocyte infiltration after LPS-induced inflammation. WT and a7-K0 mice will be randomly divided into 3 treatment groups: 1) LPS/saline, 2) LPS/vehicle, and LPS/4R and sacrificed at 24hrs, 72hrs, and 1 week after LPS injection. Flow cytometry analysis will be performed to calculate the average percentage of M1- M2 infiltrated monocytes over time. 3) To determine if 4R-cembranoid treatment improves memory deficits associated with LPS-induced inflammation in mice and whether the effect is a7 receptor dependent. Mice will be randomly divided into the same 3 treatment groups as in Aim 2. Memory function will be assessed using the novel object recognition test. 4R hippocampal neuroprotection will be determined by immunohistochemistry using stereological counting of NeuN-positive neurons in brain slices. The results of these studies will allow us to determine the mechanism of action of 4R to reduce inflammation and improve behavioral function in brain following LPS-induced inflammation in mice.

神经炎症是脑缺血的重要特征。但是，持久 在恢复阶段，炎症会导致进行性组织损伤。 细菌脂多糖内毒素（LPS）已被广泛用作全身模型 腹膜内LPS向动物注射已显示出产生 大脑中的炎症信号导致促炎细胞因子的产生增加 和记忆防御。 A7乙酰胆碱受体的激活（A7）已被证明可以减少 体内促炎细胞因子的释放。血清TNF-α水平明显更高 在LPS内毒素后两个小时，在A7敲除小鼠（A7-K0）中，相比之下。在 创伤性脑损伤模型，A7-K0小鼠的血浆水平明显较高 受伤后24小时时的1B小鼠比WT小鼠。我们的研究小组已经确定了天然的杜松子素， 4R，具有神经保护特性。 4R预防体内神经元毒性 在缺血性中风的啮齿动物模型中，大大降低了梗塞大小。我们的初步数据 显示4R治疗在小鼠中显着降低血清TNF-α，IL-1B和脑MCP-1水平 LPS引起的炎症之后。我们的假设指出4R治疗下降了LPS- 诱导神经炎症并通过增强小鼠的记忆功能 A7受体。以下具体目的旨在检验以下假设：1）调查 如果4R降低小鼠的神经炎症以及该作用是否由A7介导 接收者。将通过注射LPS，然后是1）盐水，2）载体或车辆来检查4R的效果 3）4R。抗炎细胞因子以及AKT，NF-KB和CREB的磷酸化 信号蛋白将在脑皮质和海马匀浆中测量。 2）到 检查LPS诱导后4R和A7受体在单核细胞浸润中的作用 炎。 WT和A7-K0小鼠将随机分为3个治疗组：1） LPS/Saline，2）LPS/车辆和LPS/4R，在24小时，72小时和LPS后1周处死 注射。流式细胞仪分析将进行计算M1-的平均百分比 M2随着时间的推移浸润的单核细胞。 3）确定4R-螺旋体治疗是否有所改善 记忆定义与LPS诱导的小鼠注射相关的记忆以及是否 效果取决于A7接收器。小鼠将随机分为相同的3疗法 AIM 2中的组将使用新颖的对象识别测试评估内存功能。 4R海马神经保护将通过使用免疫组织化学确定 脑切片中新阳性神经元的立体学计数。这些研究的结果将 允许我们确定4R的作用机理减少炎症并改善 LPS引起的小鼠炎症后大脑的行为功能。