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nicotinic cholinergic receptors in esophagus:Early alteration during carcinogenesis and prognostic value

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World Journal of Gastroenterology 2016年第31期22卷 7146-7156页

作者： Marina Chianello Nicolau Luis Felipe Ribeiro Pinto Pedro Nicolau-Neto Paulo Roberto Alves de Pinho Ana Rossini Tatiana de Almeida Simao Sheila Coelho Soares Lima Departamento de Bioquímica Instituto de Biologia Roberto Alcantara GomesUniversidade do Estado do Rio de Janeiro Programa de Carcinogênese Molecular Instituto Nacional de CancerCoordenacao de Pesquisa Gastroenterologia Hospital Universitário Pedro ErnestoUniversidade do Estado do Rio de Janeiro

AIM: To compare expression of nicotinic cholinergic receptors(CHRNs) in healthy and squamous cell carcinoma-affected esophagus and determine the prognostic ***: We performed RT-q PCR to measure the expression of CHRNs in 44 esophageal samples from healthy individuals and in matched normal surrounding mucosa, and in tumors from 28 patientsdiagnosed with esophageal squamous cell carcinoma(ESCC). Next, we performed correlation analysis for the detected expression of these receptors with the habits and clinico-pathological characteristics of all study participants. In order to investigate the possible correlations between the expression of the different CHRN subunits in both healthy esophagus and tissues from ESCC patients, correlation matrices were generated. Subsequently, we evaluated whether the detected alterations in expression of the various CHRNs could precede histopathological modifications during the esophageal carcinogenic processes by using receiver operating characteristic curve analysis. Finally, we evaluated the impact of CHRNA5 and CHRNA7 expression on overall survival by using multivariate ***: CHRNA3, CHRNA5, CHRNA7 and CHRNB4, but not CHRNA1, CHRNA4, CHRNA9 or CHRNA10, were found to be expressed in normal(healthy) esophageal mucosa. In ESCC, CHRNA5 and CHRNA7 were overexpressed as compared with patient-matched surrounding non-tumor mucosa(ESCC-adjacent mucosa; P < 0.0001 and P = 0.0091, respectively). Positive correlations were observed between CHRNA3 and CHRNB4 expression in all samples analyzed. Additionally, CHRNB4 was found to be differentially expressed in the healthy esophagus and the normalappearing ESCC-adjacent mucosa, allowing for distinguishment between these tissues with a sensitivity of 75.86% and a specificity of 78.95%(P = 0.0002). Finally, CHRNA5 expression was identified as an independent prognostic factor in ESCC; patients with high CHRNA5 expression showed an increased overall survival, in comparison with those

关键词： nicotinic cholinergic receptors Esophagus Esophageal squamous cell carcinoma Tobacco Alcohol Gene expression

Neuroprotective effect of the ethanol extract of Artemisia capillaris on transient forebrain ischemia in mice via nicotinic cholinergic receptor

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Chinese Journal of Natural Medicines 2018年第6期16卷 428-435页

作者： Huiyoung Kwon Ji Wook Jung Young Choon Lee Jong Hoon Ryu Dong Hyun Kim Department of Medicinal Biotechnology College of Health SciencesDong-A UniversityBusan 49315Republic of Korea Department of Herbal Medicinal Pharmacology College of Herbal Bio-industryDaegu Haany UniversityKyungsan 38610Republic of Korea Institute of Convergence Bio-Health Dong-A UniversityBusan 49315Republic of Korea Department of Life and Nanopharmaeeutical Sciences Kyung Hee UniversityKyung Hee UniversitySeou102447Republic of Korea Department of Oriental Pharmaceutical Science College of PharmacyKyung Hee UniversityKyung Hee UniversitySeoul 02447Republic of Korea Kyung Hee East-West Pharmaceutical Research Institute Kyung Hee UniversityKyung Hee UniversitySeoul 02447Republic of Korea

Artemisia capillaris Thunberg is a medicinal plant used as a traditional medicine in many cultures. It is an effective remedy for liver problems including hepatitis. Recent pharmacological reports have indicated that Artemisia species can exert various neurological effects. Previously, we reported a memory-enhancing effect of Artemisia species. However, the mechanisms underlying the neuroprotective effect of A. capillaris(AC) are still unknown. In the present study, we investigated the effect of an ethanol extract of AC on ischemic brain injury in a mouse model of transient forebrain ischemia. The mice were treated with AC for seven days, beginning one day before induction of transient forebrain ischemia. Behavioral deficits were investigated using the Y-maze. Nissl and Fluoro-jade B staining were used to indicate the site of injury. To determine the underlying mechanisms for the drug, we measured acetylcholinesterase activity. AC(200 mg·kg-1) treatment reduced transient forebrain ischemia-induced neuronal cell death in the hippocampal CA1 region. The AC-treated group also showed significant amelioration in the spontaneous alternation of the Y-maze test performance, compared to that in the untreated transient forebrain ischemia group. Moreover, AC treatment showed a concentration-dependent inhibitory effect on acetylcholinesterase activity in vitro. Finally, the effect of AC on forebrain ischemia was blocked by mecamylamine, a nonselective nicotinic acetylcholine receptor antagonist. Our results suggested that in a model of forebrain ischemia, AC protected against neuronal death through the activation of nicotinic acetylcholine receptors.

关键词： Artemisia capillaris Transient forebrain ischemia Acetylcholinesterase nicotinic acetylcholine receptor

Differentiation of the Agonists and Antagonists of the α7 nicotinic Acetylcholine Receptor

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Journal of Ocean University of China 2019年第5期18卷 1193-1198页

作者： WU Guanzhao XU Qingliang BAO Yilei LIU Yuwei LI Qian FANG Zhengyu FU Jingyi DING Yuhang LIANG Zhiqing JIANG Tao YU Rilei Key Laboratory of Marine Drugs Chinese Ministry of EducationSchool of Medicine and PharmacyOcean University of ChinaQingdao 266003China Laboratory for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology Qingdao 266003China

Theα7 nicotinic acetylcholine receptors(nAChRs)are widely expressed in the central and peripheral nervous systems and are important drug targets for the treatment of neurological ***,differentiation of the agonists and antagonists of the nAChR is *** this study we aimed to develop a reliable and efficient computational approach for differentiation of the agonists from the antagonists of the nAChR based on a systematical analysis of 123 ligands(87 agonists,12 partial agonists,and 24 antagonists)binding with the extracellular domain of theα7 n AChR *** results suggest that the ligand size and ligand binding affinity cannot differentiate the agonists from the antagonists of the *** ligand efficiency that considers both ligand binding affinity and size for the agonists is overall more left shifted in comparison to the antagonists,but the values of the ligand efficiency still cannot differentiate the agonists from the antagonists unless the values are either relatively high(more than-0.3 kcal mol^-1)or relatively low(less than-0.45 kcal mol^-1).Our results suggest that accurate prediction of the agonist or antagonist of the nAChR is challenging and the ligand innate configuration has to be considered as an extra for differentiation of the agonists from the antagonists of the nAChR.

关键词： nicotinic acetylcholine receptors ligand efficacy ligand binding affinity ligand efficiency

Cholinergic receptor, nicotinic, alpha 7 as a target molecule of Arctic mutant amyloid β

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Neural Regeneration Research 2018年第8期13卷 1360-1361页

作者： Naoya Sawamura Ye Ju Toru Asahi Faculty of Science and Engineering Waseda University Research Organization for Nano & Life Innovation Waseda University

Alzheimer’s disease（AD）is a progressive cognitive disorder that develops predominantly in elderly patients and is characterized by cognitive impairments affecting memory,learning,and attention（Selkoe,2002）.

关键词： Cholinergic receptor alpha 7 as a target molecule of Arctic mutant amyloid nicotinic AD

Activation of α7 nicotinic acetylcholine receptor protects against oxidant stress damage through reducing vascular peroxidase-1 in a JNK signaling-dependent manner in endothelial cells

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中国药理学通报 2015年第B11期31卷 156-157页

Aim Alpha7 nicotinic acetylcholine receptor （α7nAChR）, a subtype of nAChR regulating neurotrans- mission in central nervous system, is an essential regulator of cholinergic antiinflammatory pathway in periphery. The present study was to determine the effects of activation of α7nAChR on oxidant stress-induced injury in endo- thelial cells. Methods Cultured human umbilical vein endothelial cells were treated with H202 （400 μmol · L^-1） or H202plus PNU-282987 （ 10 μmol · L^-1 ）. Cell viability and membrane integrity were measured. AnnexinV ＋ PI assay, immunoblotting of bcl-2, bax and cleaved caspase-3, and immunofluorescence of apoptosis inducing factor （AIF） were performed to evaluate apoptosis. Protein expression of vascular peroxidase-1 （ VPO-1 ） and phosphor- JNK were measured by immunoblotting. Results Activation of α7nAChR by a selective agonist PNU-282987 pre-vented H202-indced decrease of cell viability and increase of lactate dehydrogenase release. Activation of α7nAChR markedly reduced cell apoptosis and intracellular oxidative stress level. Moreover, activation of α7nAChR reduced H2 02 -induced VPO-1 protein upregulation and JNK1/2 phosphorylation. The inhibitory effect of α7nAChR activa- tion on VPO-1 was blocked by JNK inhibitor SP600125. In addition, pretreatment of α7nAChR antagonist methyl- lycaconitine blocked the cytoprotective effect of PNU-282987. Conclusion These results provide the first evidence that activation of α7nAChR protects against oxidant stress-induced damage by suppressing VPO-1 in a JNK signa- ling pathway-dependent manner in endothelial cells.

关键词： Alpha7 nicotinic acetylcholine receptor vascular peroxidase-1 oxidation apoptosis endothelial cells JNK signaling

Effect of soman,sarin and VX on the nicotinic acetylcholine receptor

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Journal of Medical Colleges of PLA(China) 1991年第2期6卷 103-108页

作者：毛庆武朱艳萍乌乃侯苗小飞王宇 Department of Medical Protection Second Military Medical UniversityShanghai 200433

The highly specific ligand of the N-acetylcholine receptor（N-AChR）,alpha-bungarotoxin,was used to determine the effect of soman,sarin and VX onN-AChR of the diaphragm and extensor digitorum Iongus muscle of mice *** effects of the three anti-cholinesterase agents on N-AChR *** did not act directly on N-AChR and cause a change in the numberof *** decreased the binding sites of the receptor by binding with *** LD50was 0.054mg/kg in *** increased the binding sites,e.g.1～1.5 LD50soman increased the number of N-AChR of mouse diaphragm by 25%.The peak increaseof N-AChR was reached 0.5 h after poisoning and could last *** receptornumber was still 22% higher than that of the control on the fourth day aftersoman poisoning in *** mainly increased the number of extrasynapticN-AChR,leading to the enhancement of sensitivity of cholinergic effectors toacetylcholine（ACh）,which is similar to the hypersensitiveness resulting *** findings are of significance in probing the receptor mecha-nisms and treatment of soman poisoning.

关键词： nicotinic acetylcholine receptor alpha-bungarotoxin VX soman sarin

Enzyme-linked Immunosorbent Assay for Detection of Anti-idiotype Antibodies to Antibodies to Ligand of nicotinic Acetylcholine Receptor in Sera of Patients with Myasthenia Gravis

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Journal of Medical Colleges of PLA(China) 1990年第3期5卷 237-242页

作者：黄德仁涂来慧张仁琴周广智沈茜 Department of Neurology Changhai HospitalSecond Military Medical UniversityShanghai 200433 Clinical Laboratory Changhai Hospital

Anti-bungarotoxin anti-serum,which has the internal image of nicotinicacetylcholine receptor,was used as a tool to measure anti-idiotypic antibodies toantibodies to Iigand of nicotinic acctylcholine receptor in scra from 81 patients withmyasthenia ***-linked immunosorbcnt assay was *** positive ratewas 46.9%(38/81).The specific cross inhibitory test with nicotinic acetylcholinereceptor was ***-idiotype antibodies to antibodies to ligand of nicotinicacetylcholine receptor in sera of different types of myasthenia gravis patients classified ac-cording to modified Osserman’s standard and myasthenia gravis patients with or withoutthymoma were comparcd in this study and the role of anti-idiotype antibodies toantibodies to Iigand of nicotinic acctylcholinc receptor in the immunity of myasthcniagravis and the possibility of thcrapeutic use of anti-idiotype antibodies arc discussed.

关键词： myasthenia gravis enzyme-linked immunosorbent assay nicotinic acetylcholine receptor ligand antibungarotoxin antiserum anti-idiotype antibodies

Donepezil rescues the medial septum cholinergic neurons via nicotinic ACh receptor stimulation in olfactory bulbectomized mice

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Advances in Alzheimer's Disease 2013年第4期2卷 161-170页

作者： Yui Yamamoto Kohji Fukunaga Department of Pharmacology Graduate School of Pharmaceutical Science Tohoku University Sendai Japan

Olfactory bulbectomy (OBX) causes cognitive dysfunction by degeneration of cholinergic neurons in the medial septum. Here, we define an involvement of nicotinic acetylcholine receptor (nAChR) in neuroprotective effect of donepezil in the septum neurons of OBX mice. Neuroprotective effects on the medial septal cholinergic neurons were assessed after chronic donepezil administration in OBX mice. We also measured Akt and ERK phosphorylation to define the neuroprotective mechanism of donepezil. We found that treatment with donepezil (1 - 3 mg/kg) for 15 consecutive days completely rescued cholinergic neurons in the OBX mice with concomitant improved memory. Reduction of both protein kinase B (Akt) and extracellular signal-regulated kinase (ERK) phosphorylation were restored by chronic donepezil administration (1 - 3 mg/kg) in OBX mouse medial septum. Both phosphorylated Akt and ERK immunoreactivities were localized in cell bodies of choline acetyltransferase (ChAT)-positive cholinergic cells in the medial septum. Enhancement of Akt and ERK phosphorylation seen following donepezil administration was totally blocked by pre-administration of mecamylamine (10 μM), a nicotinic acetylcholine receptor antagonist. Donepezil increases phosphorylation of Akt and ERK via nAChR stimulation in the medial septum cholinergic neurons. The Akt and ERK stimulation by donepezil is associated with its ability of neuroprotection in the medial septum and memory improvement.

关键词： Donepezil Neuroprotection Medial Septum nicotinic Acetylcholine Receptors

Attenuation of nicotine-evoked Ca<sup>2+</sup>influx by antibody to the nicotinic acetylcholine receptor <i>α</i>3 subunits in human embryonic kidney cells

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Advances in Bioscience and Biotechnology 2013年第6期4卷 9-14页

作者： Shota Kobayashi Shigeru Yokoyama Takahiro Maruta Akiko Muroyama Hiroaki Yoshikawa Yasuhide Mitsumoto Department of Biophysical Genetics Kanazawa University Graduate School of Medicine Kanazawa Japan Department of Neurology and Neurobiology of Aging Kanazawa University Graduate School of Medical Science Kanazawa Japan Laboratory of Alternative Medicine and Experimental Therapeutics Department of Clinical Pharmacy Faculty of Pharmaceutical Sciences Hokuriku University Kanazawa Japan Neurological Center Kanazawa Nishi Hospital Kanazawa Japan

Autoantibody against neuronal nicotinic acetylcholine receptor (nAChR) α3 subunit is implicated in severe autonomic dysfunction in the patients with autoimmune autonomic ganglionopathy (AAG). Although this autoantibody has been revealed to impair fast excitatory synaptic transmission in autonomic ganglia, its precise mechanism remains unknown. Here, we show that antibody-induced reduction of cell-surface α3 subunits result in impairment of nicotine-evoked Ca2+ influx in stably transfected human embryonic kidney cells. These effects of the antibody were remarkably inhibited by interfering with the endocytic machinery at low-temperature. We conclude that reduction of nAChR in autonomic ganglia can be mediated by the endocytosis of α3 subunits, and resulted in autonomic failure in AAG patients.

关键词： nicotinic Acetylcholine Receptor α3 Subunit Antibody Endocytosis Ca2+ Influx Autoimmune Autonomic Ganglionopathy