牛膝多肽活性成分对大鼠短暂性脑缺血的保护作用*

何春娇**,程琼,丁***

(南通大学江苏省神经再生重点实验室神经再生协同创新中心,南通226001)

[]目的:研究牛膝多肽(Achyranthes Bidentatapolypeptides, ABPP)活性成分(activecomponent k isolated from ABPP, ABPPk)对大鼠短暂性脑缺血的保护作用。方法:体外建立原代胎鼠皮层神经元氧糖剥夺(oxygen and glucose deprivation,OGD)损伤模型,复氧复糖同时分别加入不同质量浓度ABPPk(0.04, 0.2, 1.0 μg/mL)和ABPP(1.0 μg/mL,阳性对照)处理皮层神经元,72 h后通过MTT法检测ABPPk对OGD损伤神经元活力的影响;通过Western Blot分析ABPPk对OGD损伤凋亡相关蛋白Bcl-2、Bax、Cleaved caspase-3表达的影响。体内建立大鼠短暂性大脑中动脉栓塞(transient middle cerebralartery occlusion, t-MCAO)模型,缺血2h后再灌注同时经尾静脉注射给予1.0 mg/kg ABPPk,1次/d,连续3 d。于再灌注72 h行神经行为学评分,评价ABPPk对t-MCAO大鼠神经行为缺陷的影响;再灌注72 h断头取脑,行2, 3, 5-氯化三苯基四氮唑(2, 3, 5-Triphenyl tetrazoliumchloride, TTC)染色评价ABPPk对t-MCAO大鼠脑梗死体积的影响;再灌注72 h采用末端脱氧核苷酸转移酶(terminal deoxynucleotidyltransferase, TdT)介导的dUTP生物素缺口末端标记法(TdT-mediated dUTP-biotin nickend labeling, TUNEL)试剂盒检测ABPPk对大鼠脑缺血半影区神经元凋亡的影响。结果:ABPPk在体外可有效拮抗OGD引起的皮层神经元凋亡,体内可显著减小脑梗死体积,防止神经元凋亡。结论:ABPPk对大鼠短暂性脑缺血具有神经保护作用,可为开发新型小分子活性肽类神经保护药物提供新的策略。

[关键词]脑缺血;牛膝多肽;神经保护;抗凋亡;大鼠

[中图分类号]R743.31[文献标志码] A[文章编号]1674-7887(2017)02-0091-06

Theeffects of an active component isolated from Achyranthes Bidentata polypeptideson rat transient ischemia*

HE Chunjiao**, CHENG Qiong, DING Fei***(JiangsuKey Laboratory of Neuroregeneration, Collaborative Centerof Neuroregeneration, Nantong University,Nantong 226001)

[Abstract] Objective: To determine the effects of anactive component isolated from Achyranthes Bidentata polypeptides (ABPPk) on rattransient ischemia.Methods: In vitro the primary cortexneurons from embryonic rats were subjected to oxygen and glucose deprivationinsult. Different concentration(0.04, 0.2 and 1.0 μg/mL) of ABPPk and 1.0 μg/mLABPP(served as positive control) were added into the medium respectively in themeantime of reperfusion for 72 hours. Cell viability was measured by MTT assay.Western Blot analysis was performed to examine the expression level ofapoptosis associated proteins(Bal-2, Bax and Cleaved caspase-3). A transientmiddle cerebral artery occlusion(t-MCAO) model was performed to evaluate theneuroprotective effects of ABPPk(1.0 mg/kg) in vivo. Neurological deficit wasscored at 72 h after reperfusion. Infarction volume was measured at 72 h afterreperfusion by 2, 3, 5-triphenyl tetrazolium chloride(TTC) staining and theapoptosis of neurons in ischemic penumbra was detected by terminaldeoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL) kit. Results: ABPPk could effectivelyinhibit neuronal apoptosis induced by OGD in vitro and exert neuroprotection ona rat t-MCAO model by improving neurologic deficit, decreasing infarctionvolume and inhibiting neuronal apoptosis in penumbra area. Conclusion: ABPPk could be considered as a new strategy fordeveloping a novel small molecular active peptide to remedy transient brainischemia because of its neuroprotective effects.

[Key words]brain ischemia;Achyranthes Bidentata polypeptides; neuroprotection; anti-apoptosis;rat


* [基金项目]国家自然科学基金资助项目(81501058),江苏省高校自然科学基金资助项目(15KJB310014),南通大学研究生科研创新计划资助项目(YKC15064)

** [作者简介]何春娇,女,汉族,生于1990年3月,河南省信阳市人,硕士在读,研究方向:中药与神经再生。

*** [通信作者]丁斐,电话:13862929598,E-mail: dingfei@ntu.edu.cn


NF-κB1基因多态位点rs4648127与胃癌易感性的研究*

1**,付1,廖蔼东1,何正磊2,王辰骥2,冯旭龙2,赵敦梅2,毛旭华3,乔国洪3***

(1江苏省苏州市相城区第二人民医院消化内科,苏州 2151432苏州大学基础医学与生物科学学院生物化学与分子生物学;3江苏省宜兴市人民医院检验科)

[]目的:探讨位于核因子κB1(nuclear factor kappasubunit 1, NF-κB1)基因内含子区的多态性位点rs4648127与中国人群胃癌易感性的关联。方法:采用病例-对照研究,以363例胃癌患者和506例年龄、性别相匹配的健康对照者作为研究对象,用高分辨率熔解曲线(high resolution melting, HRM)方法对rs4648127多态位点进行基因分型,最后通过SPSS 19.0统计分析软件对分型数据进行统计分析。结果:rs4648127位点的突变基因型CT与胃癌易感性有显著的相关性(P=0.005, OR=0.45,95%CI=0.25~0.79)。根据吸烟及饮酒等因素分层分析显示,在年龄组、性别组、非吸烟组及非饮酒状态组中均体现出CT基因型能够显著降低胃癌易感性风险(P<0.05)。结论:NF-κB1多态性位点rs4648127能够显著降低中国人群胃癌易感性。

[关键词]胃癌;核因子κB1;单核苷酸多态性;易感性;高分辨率熔解曲线

[中图分类号] R735.2Q343.1[文献标志码]A[文章编号]1674-7887(2017)02-0096-05

Association study betweenNF-κB1 gene polymorphism loci rs4648127 and susceptibility of gastric cancer*

JIANG Hua1**,FULin1,LIAO Aidong1,HE Zhenglei2,WANG Chenji2,FENG Xulong2,ZHAO Dunmei2,MAO Xuhua3, QIAOGuohong3***(1Department ofDigestive System, the Second People’s Hospital of Xiang Cheng District, SuzhouCity, Jiangsu Province, Suzhou 215143; 2School of Basic Medical and BiologicalScience, Biochemistry and Molecular Biology, Soochow University; 3Department ofClinical Laboratory, Yixing People’s Hospital, Jiangsu Province)

[Abstract] Objective: To investigate the association of singlenucleotide polymorphism(SNP) locus rs4648127 of(nuclear factor kappa B subunit1, NF-κB1) gene with the susceptibility of Chinese gastric cancer. Methods: In this case-control study,363 cases of gastric cancer patients and 506 healthy control were used asresearch subjects. The locus of rs4648127 wasgeno-typed by high resolution melting(HRM) method, and the genotyping data wasanalysed by SPSS 19.0 software. Results:There was a significant association between the mutation genotype CT of thers4648127 and the susceptibility to gastric cancer(P=0.005, OR=0.45,95%CI=0.25-0.79). CT genotype was significantly decreased susceptibility ofgastric cancer in the age group, gender group, non-smoking group and non-drinkingstatus group was found by stratification analysis according to smoking,drinking and other factors(P<0.05). Conclusion:The locus of rs4648127 of NF-kB1 can significantly decrease the risk ofgastric cancer in China.

[Key words]gastric cancer;nuclera factor kappa subunit 1; single nucleotidepolymorphism; susceptibility;high resolution melting


* [基金项目]苏州市相城区科技计划项目(XJ201452),2016年宜兴市科技局民生项目9号(社会发展类),无锡市卫计委面上项目(MS201536)

** [作者简介]蒋骅,男,汉族,生于1978年3月,江苏省苏州市人,硕士,副主任医师,研究方向:消化内科相关疾病。

*** [通信作者]乔国洪,电话:0510-87921355,E-mail:staff1000@yxph.com