α7烟碱型乙酰胆碱受体在胆碱能抗炎通路中的作用机制及应用现状

胆碱能抗炎通路是一条神经-免疫调节通路,以中枢神经系统的迷走神经为起点,以神经递质乙酰胆碱为基础,以免疫细胞膜表面的α7烟碱型乙酰胆碱受体(α7 nicotinic acetylcholine receptor, α7 nAChR)为关键作用位点。其中,电信号与化学信号相互转化,激活JAK-STAT3、PI3K-Akt等信号通路,抑制NF-κB的核转位,进而抑制促炎因子的释放,促进抑炎因子的释放,调节和控制机体的炎症,具体的作用机制正在研究中。研究表明,可以通过使用药物激活α7 nAChR和电刺激迷走神经这2种方法,激活胆碱能抗炎通路。其中,激活α7 nAChR具有操作简便、创伤小和效果显著等优势。常用药物有选择性激动剂,例如PNU282987和GTS-21等,非选择性激动剂例如烟碱。多项研究显示,该方法在脓毒症、缺血再灌注、胃肠炎、骨关节炎和自身免疫病等外周器官炎性疾病治疗中都具有一定的效果。α7 nAChR作为胆碱能抗炎通路的关键作用位点,成为许多炎性疾病的潜在治疗靶点,本文对胆碱能抗炎通路中的α7 nAChR所涉及的抗炎机制、通路激活方式,以及其近年来在炎性疾病中的应用进行了综述,为未来研究其具体作用机制和新的治疗靶点提供参考。     

炎症反射的研究进展

以往认为体液调节机制是防止多种促炎因子过量产生的唯一通路,但本世纪伊始新发现了一条生理性的神经性抗炎通路,即炎症反射,而迷走神经则是这一反射的重要载体,免疫细胞上表达的α7nAChR是该反射的"效应器",中枢神经系统也参与了该反射的信号收集、整合与调节的过程.由于这一通路的抗炎作用具有高效、特异性强等特点,其对于治疗炎性疾病具有极其重要的临床应用前景.本文对炎症反射的最新研究进展予以综述.     

IFITM3在脓毒症模型及胆碱能抗炎模型中的表达

目的:探讨干扰素诱导的跨膜转运蛋白3(IFITM3)在LPS刺激的RAW264.7细胞系的脓毒症模型中的表达以及胆碱能抗炎模型中的表达。方法:用1μg/mL LPS刺激RAW264.7细胞24、48、72 h后,用Western-Blot法检测各组细胞IFITM3蛋白表达水平。用1μg/mL LPS刺激RAW264.7细胞后,给予50μM胆碱能受体激动剂GTS-21以及同时给予100 n M胆碱能受体拮抗剂α-BGT刺激细胞24 h后,用Western-Blot法检测各组细胞IFITM3蛋白表达水平。用ELISA法检测IL-1β的方法验证脓毒症模型和胆碱能抗炎模型的建立。结果:(1)1μg/mL LPS刺激RAW264.7细胞后,IFITM3蛋白表达明显降低(P0.01)。(2)1μg/mL LPS刺激RAW264.7细胞后再给予50μM GTS-21,IFITM3蛋白表达明显升高(P0.001);而给予100 nMα-BGT后,IFITM3蛋白表达明显降低(P0.001)。结论:LPS刺激的RAW264.7细胞IFITM3蛋白表达降低。给予胆碱能激动剂GTS-21后能够逆转LPS诱导的IFITM3表达的降低,给予胆碱能受体拮抗剂α-BGT则能阻断这种现象。IFITM3有可能在脓毒症中发挥保护作用,并且参与了胆碱能抗炎通路抗炎过程的调节。     

模式识别受体介导炎症信号通路及人参属植物皂苷抗炎研究进展

炎症是一种非常常见而又十分重要的基本病理过程,临床表现为红、肿、痛,其伴随着疾病的发生而产生,同时又会加重疾病的发展,有些甚至会产生恶性肿瘤。在炎症分子机制的研究中,已发现多条重要的信号通路,其中模式识别受体介导的炎症信号通路在很多疾病中都扮演着重要的角色,基于这些重要的炎症信号通路中的关键分子筛选抗炎药物受到广泛关注。人参属植物是名贵的中药材,具有很高的药用价值。我国人参属植物主要包括:人参、三七、竹节参及其变种珠子参等。皂苷是人参属植物的主要活性成分之一,药理研究显示其具有良好的抗炎效果。本文就此将近年来模式识别受体介导的炎症信号通路及人参属植物皂苷抗炎的研究成果进行总结,为抗炎药物研究提供理论基础。     

α7烟碱型胆碱能受体调节胆管癌细胞化疗敏感性的相关探讨

目的：观察尼古丁对胆管癌细胞QBC939化疗敏感性影响,并初步探讨其作用靶点。方法：应用MTT法检测α7烟碱型胆碱能受体激动剂尼古丁及其阻断剂仪银环蛇毒素（α-BTX）干预后的人胆管癌细胞QBC939经5-FU处理后存活增殖能力变化,应用单克隆平板试验观察尼古丁及α银环蛇毒素对5-FU处理后细胞克隆形成率变化。结果：经5-fu处理后,尼古丁刺激组（终浓度分别为10-3g／L、10-4g／L、10-5g／L）细胞存活率分别为128％、124％、118％,细胞存活率较阴性对照组明显升高,并呈一定的浓度依赖性,而α银环蛇毒素刺激组（2ug／mL）、尼古丁α银环蛇毒素联合组细胞存活率分别为92％、94％、93％、92％。尼古丁刺激组（6．2±0．40）的克隆形成能力明显高于α银环蛇毒素刺激组（3．2±0．20）、联合组（3．2±0．20）及对照组（3．4±0．33）;结论：尼古丁可明显降低胆管癌细胞对化疗药物的敏感性,其可能是通过α7烟碱型胆碱能受体发挥化疗抵抗效应。     

乙酰胆碱信号通路在肺癌中的作用

肺癌作为当今世界最为致命的疾病之一,严重地威胁着人类的生命健康。神经递质乙酰胆碱(acetylcholine,ACh)作为肺癌的自分泌和旁分泌生长因子,可与肺癌细胞上的烟碱型乙酰胆碱受体(nicotinic acetylcholine receptor,nAChRs)和毒蕈碱型乙酰胆碱受体(muscarinic acetylcholine receptors,mAChRs)结合,促进肺癌细胞的增殖、迁移和侵袭。近年来,全基因组数据研究表明,nAChRs与吸烟相关肺癌风险密切相关,因此其在肺癌发生发展中的作用机制的研究对肺癌药物的研发具有重要意义。本文对乙酰胆碱信号通路在肺癌进展中的作用机制进行综述,为其成为肺癌治疗新分子靶点和药物的开发提供依据。     

α7烟碱型乙酰胆碱受体在烟碱成瘾不同阶段发挥的作用及其作用机制

烟碱是烟草中备受关注的致瘾性物质,是吸烟者持续使用烟草制品的主要原因之一.Alpha7烟碱型乙酰胆碱受体(alpha7 nicotinic acetylcholine receptor,α7nAChR)在中枢神经系统广泛存在,其功能涵盖学习记忆、认知障碍、神经退行性疾病和药物成瘾等多个方面.近年来,大量研究报道了调控α...     

非神经元型胆碱能系统在（前）脂肪细胞的表达及nAChRα7功能状态对前脂肪细胞Visfatin基因表达的影响

原核生物、真核生物、植物体的非神经细胞和组织中,尤其是多种免疫活性细胞中,均证实乙酰胆碱酯酶（acetylcholinesterase,AChE）、胆碱乙酰转移酶（choline acetyltransferase,ChAT）和乙酰胆碱受体（acetylcholine receptor, AChR）各亚型在内的胆碱能系统组分的存在,其中烟碱样乙酰胆碱受体α7（nicotinic acetylcholine receptor α7,nAChRα7）是烟碱样胆碱能抗炎通路（nicotinic anti-inflammatory pathway）中重要的分子核心机制,同时也是机体限制宿主防御反应扩大的内源性抗炎机制之一. 本文旨在探讨（前）脂肪细胞上非神经元型胆碱能系统是否存在及初步揭示烟碱样胆碱能受体α7对前脂肪细胞功能的影响. 以体外培养的3T3-L1前脂肪细胞为研究对象,采用免疫组化和蛋白质免疫印迹技术,分别检测前脂肪细胞和成熟脂肪细胞中乙酰胆碱酯酶、胆碱乙酰转移酶和烟碱样乙酰胆碱受体α7的3种胆碱能系统主要组分的蛋白表达. 另将前脂肪细胞分为给予广谱烟碱样乙酰胆碱受体激动剂尼古丁、特异性烟碱样乙酰胆碱受体α7激动剂氯化胆碱及特异性烟碱样乙酰胆碱受体α7拮抗剂甲基牛扁亭碱干预12 h、24 h、36 h,并设立相应处理时间的空白对照组,逆转录聚合酶链反应检测前脂肪细胞visfatin mRNA表达情况. 免疫组化染色可见前脂肪细胞中AChE、ChAT及AChRα7均有阳性表达;蛋白免疫印迹检测进一步半定量证实了前脂肪细胞和成熟脂肪细胞中AChE、ChAT及AChRα7的蛋白表达;拮抗剂甲基牛扁亭碱(10⁶～10⁴mol/L)时间、剂量依赖性上调前脂肪细胞visfatin mRNA表达(1.3～1.55fold,P<0.01),与对应空白对照组相比,存在显著性统计学差异; 加入不同剂量的尼古丁和氯化胆碱,则前脂肪细胞中visfatin mRNA表达水平与对应空白对照组相比,均不同程度地下降,其中以氯化胆碱的抑制效应更为显著. 前脂肪细胞与成熟脂肪细胞中均存在有独立的胆碱能体系,其中AChRα7很可能在调节脂肪细胞因子分泌及肥胖相关的病理生理过程中发挥重要作用.     

电针对术后认知功能障碍大鼠认知功能及海马α7nAChR受体表达的影响

摘要 目的：探讨电针对术后认知功能大鼠认知功能及海马α7型神经烟碱胆碱能受体(nicotinic acetylcholine receptor,α7nAChR)表达含量的影响。方法：选择120只雄性SD大鼠,将其随机分为以下5组：对照组(Ctrl组)、手术组(Op组)、电针组(EA)、α7nAChR抑制剂 (EA+α-BGT)组、α7nAChR激动剂 (PHA-543,613组),每组12只,每组又分为1 d和3 d两个亚组。采用Morris水迷宫检测认知功能,ELISA检测血清肿瘤坏死因子(Tumor necrosis factor,TNF)和白细胞介素-1β(interleukin-1β,IL-1β)、高迁移率族蛋白1(High mobility group box 1,HMGB-1)含量,RT-PCR检测海马TNF-α和IL-1β、HMGB-1 m-RNA表达,蛋白印迹法检测海马α7nAChR表达,甲苯胺蓝法检测海马CA1区肥大细胞活化情况,Tunel法检测海马CA1区细胞凋亡情况。结果：与Op组相比,EA组、PHA-543,613组术后第1 d、第3 d逃避潜伏期显著缩短,穿越平台次数增加 (P<0.05);与EA组相比,EA+α-BGT组术后第1 d、第3 d逃避潜伏期显著延长,穿越平台次数明显减少(P<0.05)。与Op组相比,EA组、PHA-543,613组鼠术后第1 d、第3 d血清TNF-α和IL-1β、HMGB-1含量显著降低,海马TNF-α和IL-1β、HMGB-1 m-RNA表达亦明显下调,海马α7nAChR蛋白含量表达上调、海马CA1区肥大细胞活化数目、Tunel阳性细胞数目减少,两组比较差异有统计学意义(P<0.05)。与EA组相比,EA+α-BGT组大鼠术后第1 d、第3 d血清TNF-α和IL-1β、HMGB-1含量增加,海马TNF-α和IL-1β、HMGB-1 m-RNA表达亦明显上调,海马α7nAChR蛋白表达上调,海马CA1区肥大细胞活化数目、Tunel阳性细胞数目亦明显增加(P<0.05)。结论：电针可能通过抑制中枢肥大细胞活化,上调脑内α7nAChR蛋白表达,抑制TNF-α和IL-1β、HMGB-1表达和释放,进而改善胫骨骨折术后大鼠学习与记忆能力。     

Neuro-immune interactions via the cholinergic anti-inflammatory pathway

The overproduction of TNF and other cytokines is associated with the pathophysiology of numerous diseases. Controlling cytokine synthesis and release is critical for preventing unrestrained inflammation and maintaining health. Recent studies identified an efferent vagus nerve-based mechanism termed "the cholinergic anti-inflammatory pathway" that controls cytokine production and inflammation. Here we review current advances related to the role of this pathway in neuro-immune interactions that prevent excessive inflammation. Experimental evidence indicates that vagus nerve cholinergic anti-inflammatory signaling requires alpha7 nicotinic acetylcholine receptors expressed on non-neuronal cytokine-producing cells. Alpha7 nicotinic acetylcholine receptor agonists inhibit cytokine release and protect animals in a variety of experimental lethal inflammatory models. Knowledge related to the cholinergic anti-inflammatory pathway can be exploited in therapeutic approaches directed towards counteracting abnormal chronic and hyper-activated inflammatory responses.     

The cholinergic anti-inflammatory pathway: a missing link in neuroimmunomodulation

This review outlines the mechanisms underlying the interaction between the nervous and immune systems of the host in response to an immune challenge. The main focus is the cholinergic anti-inflammatory pathway, which we recently described as a novel function of the efferent vagus nerve. This pathway plays a critical role in controlling the inflammatory response through interaction with peripheral a7 subunit-containing nicotinic acetylcholine receptors expressed on macrophages. We describe the modulation of systemic and local inflammation by the cholinergic anti-inflammatory pathway and its function as an interface between the brain and the immune system. The clinical implications of this novel mechanism also are discussed.     

Nicotine protects kidney from renal ischemia/reperfusion injury through the cholinergic anti-inflammatory pathway

Kidney ischemia/reperfusion injury (I/R) is characterized by renal dysfunction and tubular damages resulting from an early activation of innate immunity. Recently, nicotine administration has been shown to be a powerful inhibitor of a variety of innate immune responses, including LPS-induced toxaemia. This cholinergic anti-inflammatory pathway acts via the alpha7 nicotinic acetylcholine receptor (alpha7nAChR). Herein, we tested the potential protective effect of nicotine administration in a mouse model of renal I/R injury induced by bilateral clamping of kidney arteries. Renal function, tubular damages and inflammatory response were compared between control animals and mice receiving nicotine at the time of ischemia. Nicotine pretreatment protected mice from renal dysfunction in a dose-dependent manner and through the alpha7nAChR, as attested by the absence of protection in alpha7nAChR-deficient mice. Additionally, nicotine significantly reduced tubular damages, prevented neutrophil infiltration and decreased productions of the CXC-chemokine KC, TNF-alpha and the proinflammatory high-mobility group box 1 protein. Reduced tubular damage in nicotine pre-treated mice was associated with a decrease in tubular cell apoptosis and proliferative response as attested by the reduction of caspase-3 and Ki67 positive cells, respectively. All together, these data highlight that nicotine exerts a protective anti-inflammatory effect during kidney I/R through the cholinergic alpha7nAChR pathway. In addition, this could provide an opportunity to overcome the effect of surgical cholinergic denervation during kidney transplantation.     

Effect of dexmedetomidine on rats with convulsive status epilepticus and association with activation of cholinergic anti-inflammatory pathway

Convulsive status epilepticus (CSE) is a neurological disease with contraction and extension of limbs, leading to damage of hippocampus and cognition. This study aimed to explore the effects of dexmedetomidine (DEX) on the cognitive function and neuroinflammation in CSE rats. All rats were divided into control group, CSE group and DEX group. Morris water maze test was used to measure cognitive function. Acute hippocampal slices were made to detect long-term potentiation (LTP). Immunohistochemistry was used to determine the expression of α7-nicotinic acetylcholine receptor (α7-nAChR) and interleukin-1β (IL-1β). Enzyme-linked immunosorbent assay (ELISA) was used to measure serum levels of IL-1β, tumor necrosis factor-α (TNF-α), S-100β and brain-derived neurotrophic factor (BDNF). Our results showed that DEX improved the memory damage caused by CSE. DEX reduced seizure severity and increased the amplitudes and sustainable time of LTP, and also inhibited the hippocampal expression of α7-nAChR and IL-1β in CSE rats. DEX treatment decreased serum IL-1β, TNF-α and S-100β levels and increased BDNF levels. The effects of DEX on seizure severity and LTP could be simulated by nicotine or attenuated by concurrent α-bungarotoxin (α-BGT) treatment. In conclusions, DEX significantly improved spatial cognitive dysfunction, reduced seizure severity and increased LTP in CSE rats. Improvements by DEX were closely related to enhancement of cholinergic anti-inflammatory pathway.     

Lipopolysaccharide-induced Notch signaling activation through JNK-dependent pathway regulates inflammatory response

Background  

Notch and TLR pathways were found to act cooperatively to activate Notch target genes and to increase the production of TLR-induced cytokines in macrophages. However, the mechanism of LPS-induced Notch activation and its role in sepsis still remains unclear.     

Metformin inhibits inflammatory response via AMPK-PTEN pathway in vascular smooth muscle cells

Atherosclerosis is a chronic inflammation of the coronary arteries. Vascular smooth muscle cells (VSMCs) stimulated by cytokines and chemokines accelerate the inflammatory response and migrate to the injured endothelium during the progression of atherosclerosis. Activation of AMP activated protein kinase (AMPK), a key sensor maintaining metabolic homeostasis, suppresses the inflammatory response. However, how AMPK regulates the inflammatory response is poorly understood. To identify the mechanism of this response, we focused on phosphatase and tensin homolog (PTEN), which is a negative regulator of inflammation. We investigated that activation of AMPK-induced PTEN expression and suppression of the inflammatory response through the AMPK-PTEN pathway in VSMCs. We treated with the well-known AMPK activator metformin to induce PTEN expression. PTEN was induced by metformin (2mM) and inhibited by compound C (10μM) and AMPK siRNA. Tumor necrosis factor-alpha (TNF-α) was used to induce inflammation. The inflammatory response was confirmed by cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS) expression, and activation of nuclear factor (NF)-κB. Metformin suppressed COX-2 and iNOS mRNA and protein expression dose dependently. Treatment with compound C and bpv (pic) in the presence of metformin, iNOS and COX-2 protein expression increased. NF-κB activation decreased in response to metformin and was restored by inhibiting AMPK and PTEN. Inhibiting AMPK and PTEN restored ROS levels stimulated with TNF-α. Taken together, PTEN could be a possible downstream regulator of AMPK, and the AMPK-PTEN pathway might be important in the regulation of the inflammatory response in VSMCs.     

The effect of anti-inflammatory properties of ferritin light chain on lipopolysaccharide-induced inflammatory response in murine macrophages

Ferritin light chain (FTL) reduces the free iron concentration by forming ferritin complexes with ferritin heavy chain (FTH). Thus, FTL competes with the Fenton reaction by acting as an antioxidant. In the present study, we determined that FTL influences the lipopolysaccharide (LPS)-induced inflammatory response. FTL protein expression was regulated by LPS stimulation in RAW264.7 cells. To investigate the role of FTL in LPS-activated murine macrophages, we established stable FTL-expressing cells and used shRNA to silence FTL expression in RAW264.7 cells. Overexpression of FTL significantly decreased the LPS-induced production of tumor necrosis factor alpha (TNF-α), interleukin 1β (IL-1β), nitric oxide (NO) and prostaglandin E2 (PGE2). Additionally, overexpression of FTL decreased the LPS-induced increase of the intracellular labile iron pool (LIP) and reactive oxygen species (ROS). Moreover, FTL overexpression suppressed the LPS-induced activation of MAPKs and nuclear factor-κB (NF-κB). In contrast, knockdown of FTL by shRNA showed the reverse effects. Therefore, our results indicate that FTL plays an anti-inflammatory role in response to LPS in murine macrophages and may have therapeutic potential for treating inflammatory diseases.