专题3:抗炎症和免疫药理学

专题3:抗炎症和免疫药理学

T3-1

人参地黄桑葚片对小鼠免疫功能的影响

付 萌，洪铁

（吉林大学药学院药理教研室，吉林长春 130021）

目的探索人参地黄桑葚片对小鼠免疫功能的影响。方法将三批雄性ICR小鼠每批随机分为对照组、人参地黄桑葚片高剂量组（2.250 g·kg-1）、人参地黄桑葚片中剂量组（0.750 g·kg-1）、人参地黄桑葚片低剂量组（0.375 g·kg-1），每组15只，连续灌胃给药1个月后，称量并计算小鼠免疫器官指数（胸腺和脾脏），采用绵羊红细胞（SRBC）诱导法测定小鼠迟发型变态反应（DTH）；采用乳酸脱氢酶（LDH）法测定自然杀伤（NK）细胞的活性、MTT法测定刀豆蛋白A（ConA）诱导的脾淋巴细胞增殖能力；采用滴片法测定腹腔巨噬细胞对鸡红细胞（CRBC）的吞噬作用。结果人参地黄桑葚片高、中、低剂量组均能显著增加小鼠胸腺指数及脾指数（P＜0.05）；人参地黄桑葚片高、中剂量组能显著增强小鼠NK细胞活性及SRBC诱导的小鼠DTH（P＜0.05）；人参地黄桑葚片高剂量组能显著提高ConA诱导的脾淋巴细胞的增殖能力及腹腔巨噬细胞对CRBC的吞噬作用（P＜0.05）。结论人参地黄桑葚片能够显著增强小鼠的免疫功能。

人参地黄桑葚片；免疫功能

T3-2

利用斑马鱼模型研究姜黄素抗炎的分子机制

王荣春，韩利文，何秋霞，陈锡强，刘可春

（山东省科学院生物研究所，山东省生物传感器重点实验室，山东省科学院药物筛选技术重点实验室，山东省生物检测技术工程实验室，山东济南 250014）

摘要：目的利用斑马鱼炎症模型探讨姜黄素抗炎作用，分析姜黄素抗炎的分子机制。方法采用3dpf（days after fertil⁃ization）转基因斑马鱼Tg（Lyz：GFP）幼鱼，用姜黄素0，5，10，30和50 μmol·L-1处理1 h后，通过用硫酸铜20 μmol·L-1处理造成急性炎症，体式显微镜下观察姜黄素对中性粒细胞的迁移、定位，并统计迁移中性粒细胞的数量；通过脂多糖（LPS）10 μmol·L-1处理受精后24 h野生型斑马鱼AB品系造成炎症模型，并姜黄素0，5，10，30和50 μmol·L-1，通过NO和ROS检测试剂盒检测NO和ROS的表达水平，通过荧光倒置显微镜进行拍照，统计荧光强度。结果在硫酸铜造成的转基因斑马鱼急性炎症模型下，与对照处理相比，姜黄素30和50 μmol·L-1抑制硫酸铜造成的中心粒细胞向斑马鱼测线的渗漏，渗漏的中性粒细胞数目明显减少（P＜0.05）；LPS和姜黄素处理斑马鱼幼鱼，姜黄素30和50 μmol·L-1可以降低LPS处理诱导的NO和ROS的表达水平（P＜0.05）。结论本研究通过不同斑马鱼炎症模型验证了姜黄素具有抗炎的作用，其机制是通过抑制NO和ROS信号的释放，减少中性粒细胞的渗漏，从而缓解炎症。

关键词：姜黄素；斑马鱼；炎症；硫酸铜；脂多糖；

基金项目：国家自然科学基金青年基金项目（31400979）；山东省科学院青年基金项目（2015QN010）

通讯作者：刘可春，E-mail：hliukch@sdas.org，Tel：（0531）82605352

T3-3

双环醇抗炎作用机制研究

刘 宁，李芳芳，张 丹，鲍秀琦，孙 华

（中国医学科学院/北京协和医学院药物研究所，北京100050）

摘要：目的研究双环醇抗炎机制及可能靶点。方法佛波酯（PMA）诱导分化THP-1单核细胞成巨噬细胞，LPS刺激。ELISA检测细胞上清中炎症因子含量，Western蛋白印迹法检测蛋白表达。siRNA进行基因沉默，进一步分析炎症因子及蛋白表达的变化。3T3-L1前脂肪细胞分化为脂肪细胞，油红O染色。结果双环醇对LPS刺激的人巨噬细胞炎症因子TNF-α、IL-6、IL-1β的释放表现出显著的抑制活性，其能抑制PI3K的表达，促进IκB表达，抑制NF-κB入核，但对Akt、IKK磷酸化无明显影响。进一步的研究显示，双环醇显著促进PPARγ表达，沉默PPARγ基因，双环醇对IκB的促表达作用减弱，抑制炎症因子分泌的作用降低。油红O染色结果显示，双环醇可促进前脂肪细胞的分化，但作用弱于罗格列酮。结论双环醇具有良好的抗炎活性，其抗炎作用与促进PPARγ表达，进而促进IκB表达，抑制NF-κB入核有关。

关键词：双环醇；LPS；细胞因子；PPARγ；IκB；

基金项目：国家科技计划863项目（2014AA021101）；国家自然科学基金项目（81173073）

通讯作者：孙 华，E-mail：sunhua@imm.ac.cn，Tel：13811929788

T3-4

虾青素对博来霉素致大鼠肺纤维化的干预作用及机制

王芳芹，刘 涛，张 攀，阳群芳，陈晓红

（第三军医大学药学系药理教研室，重庆 400038）

摘要：目的探讨虾青素对博莱霉素所致SD大鼠肺纤维化的干预作用。方法将SD大鼠随机分为假手术组、虾青素组、博来霉素组、虾青素+博来霉素组。经气道注入博莱霉素（5 mg·kg-1）或生理盐水后，每天灌服虾青素（2 mg·kg-1）或植物油连续14 d，最后一次给药24 h后取材。计算肺系数，进行HE染色及Masson染色，显微镜下观察大鼠肺部炎症及纤维化程度；酶联免疫夹心法检测血清中TGF-β1水平；免疫组化观察肺间质α-SMA蛋白表达变化；碱水解法检测羟脯氨酸含量；Western蛋白印迹法法检测ERK1/2及Smad2/3蛋白磷酸化水平。结果与假手术组相比，博来霉素组肺损伤严重，有明显的肺纤维化病灶形成；与模型组比较，药物治疗组肺损伤明显减轻，胶原蛋白沉积减少，羟脯氨酸含量也明显降低（P＜0.01），α-SMA蛋白表达减少，TGF-β1水平明显降低（P＜0.05），同时ERK1/2蛋白及Smad2/3蛋白磷酸化水平明显下降。结论虾青素能明显改善肺纤维化，其作用机制可能与降低TGF-β1水平，抑制ERK1/2蛋白及Smad2/3蛋白磷酸化有关。

关键词：虾青素；肺纤维化；博来霉素

基金项目：国家自然科学基金项目（81472912）

通讯作者：陈晓红，E-mail：pharsunr@126.com，Tel：（023）68752364

T3-5

炎症免疫反应软调节—药物研究新方向

魏伟

（安徽医科大学临床药理研究所，抗炎免疫药物教育部重点实验室，抗炎免疫药物安徽省协同创新中心，安徽合肥230032）

摘要：炎症免疫反应（IIR）是机体炎症免疫相关组织、器官和细胞依据内外环境变化所表现出的适度或异常的系统反应，几乎涉及机体大多系统的保护或病理反应。参与IIR的细胞不仅涉及常见的炎症免疫细胞（如巨噬细胞、树突细胞、T细胞、B细胞等及其亚型），而且涉及非传统的炎症免疫细胞（如胶质细胞、内皮细胞、上皮细胞、成纤维细胞、滑膜细胞、肝细胞等），它们有着类似的或共性的细胞信号转导。目前临床使用的影响IIR药物虽然在治疗中发挥了较好疗效，但出现的不良反应尤其是肝、肾、胃肠、恶性感染、诱发肿瘤等严重毒性令人困惑，这些药物通过不同机制在抑制IIR所致病理反应的同时，也损害了组织、器官和细胞的生理功能，尤其对于需要长期用药的慢性复杂性疾病更是如此。炎症免疫反应软调节（SRIIR）是指药物在控制IIR相关细胞异常活化的同时尽量减少对细胞生理功能的损害，即：行驶SRIIR的药物不应是对细胞功能、基因和蛋白表达或活性的完全抑制，而是在不同细胞上发现共性的或类似的信号转导和网络调控关键分子及特征，选择性地将异常改变的细胞和分子活性调控至正常生理水平，恢复细胞功能的动态平衡，以发挥药物的治疗作用，且尽量减少不良反应。SRIIR是研发治疗IIR相关疾病创新药物的新方向。

关键词：炎症免疫反应；炎症免疫反应软调节；信号转导；新药发现

基金项目：国家自然科学基金重点项目资助（81330081）

通讯作者：魏 伟，E-mail：wwei@ahmu.edu.cn，Tel：（0551）65161209

T3-6

二仙膏对化疗后白细胞减少症的药效作用

许律捷1，康 德1，王金华1，刘艾林1，杜冠华1，杜新磊2

（1.中国医学科学院北京协和医学院药物研究所，北京100050；2.山东方健制药有限公司）

摘要：目的研究探讨传统中药二仙膏对环磷酰胺致小鼠白细胞减少症的影响。方法①在治疗实验 昆明小鼠随机分为对照组、模型组（环磷酰胺，100 mg·kg-1，ip）、二仙膏低、中、高三个剂量组（3，6，12 g·kg-1，ig）组、阳性药组（瑞白，50 μg·kg-1，ih）。检测小鼠的外周血白细胞数，股骨有核细胞数，计算脾脏指数和胸腺指数。在预防实验中，昆明小鼠随机分为对照组、模型组（环磷酰胺，200 mg·kg-1，ip）、二仙膏低、中、高三个剂量组（1.5，3，6 g·kg-1，ig）组、阳性药组（瑞白，25 μg·kg-1，ih），对预防给药5 d，造模后给药7 d，检测小-鼠外周血白细胞数，取材后检测股骨有核细胞数，并计算脾脏指数和胸腺指数。结果在治疗实验中，与模型组相比较，二仙膏可以显著提高脾指数，并显著增加骨髓有核细胞数。②在预防实验 二仙膏可以显著增加外周血白细胞数和股骨有核细胞数，并显著提高胸腺指数。结论二仙膏对于环磷酰胺致小鼠白细胞减少症具有明显的预防和治疗作用。

关键词：二仙膏；环磷酰胺；白细胞减少症

基金项目：山东省科技重大专项项目（2015ZDXX0302A01）

通讯作者：刘艾林，E-mail：liuailin@imm.ac.cn，Tel：（010）63165184；杜冠华，E-mail：dugh@imm.ac.cn，Tel：（010）63165184；杜新磊，E-mail：Sdfjzy@163.com，Tel：（0537）2315804

T3-7

不同提取方法及给药途径的肠腑康胶囊对大鼠脾虚型溃疡性结肠炎的影响

张小丽，陈瑞明，李 芳，杨智锋，樊炳辉

（陕西省中医药研究院，陕西西安 710003）

摘要：目的探究不同提取方法及给药途径的肠腑康胶囊对大鼠脾虚型溃疡性结肠炎（UC）的影响。方法选取正常大鼠84只，随机分成6组：阴性对照组，模型组，口服给药+直肠给药组，分别提取口服给药组，混合提取口服给药组，补脾益肠丸阳性组。对照组正常饲养外，其余6组建立脾虚型溃疡性结肠炎模型，将大鼠禁食不禁水12 h，灌服番泻叶浸剂，1/d，灌服1 d后，禁食不禁水36 h，麻醉后用0.5%肥皂水2 mL/只灌肠冲洗；动物苏醒后，正常饲养，从第2天开始，继续灌服番泻叶浸剂2 d。在已复制的脾虚型模型基础上，采用2，4，6-三硝基苯磺酸复制溃疡性结肠炎模型：将大鼠禁食24 h后，用10%水合氯醛（350 mg·kg-1）腹腔注射麻醉，固定在鼠解剖台上，准备一直径2.0 mm长约12 cm的橡胶输液管，用液体石蜡润滑后，轻缓插入肛门约8 cm，将50%乙醇与三硝基苯磺酸（TNBS，100 mg·kg-1）混合液0.25 mL注入大鼠结肠后注入0.3 mL空气，再将大鼠提尾倒置5 min，让其平躺自然清醒。于造模完成24 h后，在模型对照组中随机选取2只大鼠进行解剖，肉眼观察结肠黏膜组织损伤情况，并截取约1 cm左右病变结肠行病理组织学检查，以确定模型复制是否成功。造模成功后，模型组和阴性对照组灌服水外，剩下4组按药物剂量组别分别进行灌胃，15 d后，观察大鼠症状，体征，生理病变。结果阴性对照组大鼠结肠光滑，无水肿及血丝，与阴性对照组比较，造模后大鼠出现嗜睡，懒动，蜷缩，饮少，纳呆，精神萎靡，毛发疏松粗糙晦暗无光泽，腹泻及黏液血便症状，大鼠体质量明显下降，摄食饮水量明显降低，小便量明显减少，大便湿重增加，自发活动次数明显减少。造模后各组大鼠结肠黏膜组织在光镜下可见到大量红血丝、水肿、部分结肠溃疡。给药15 d后，解剖见给药各组大鼠结肠水肿溃疡数量明显比模型组减少，其中口服+直肠给药组结肠病变改善最为良好，水肿数量很少，溃疡数及溃疡面积最少，分别提取组次之，混合提取组不理想。病理组织学检查：阴性组结肠大体正常，大肠腺体规则，可见大量胞质富含黏液的杯状细胞，黏膜固有层、黏膜下层未见炎性细胞浸润。模型组肠黏膜可见溃疡，可见“陷窝脓肿”、“陷窝溃疡”形成，炎症程度较重，可见肠黏膜和黏膜下组织大量炎细胞浸润。阳性组结肠大体正常，大肠腺体规则，肠黏膜可见少量炎细胞浸润。口服+直肠组结肠大体正常，大肠腺体规则，肠黏膜可见少量炎细胞浸润。分别提取肠黏膜可见炎细胞浸润。混合提取肠黏膜可见轻度溃疡，可见肠黏膜和黏膜下组织大量炎细胞浸润。结论不同给药途径和提取方法的肠腑康胶囊对大鼠脾虚型溃疡性结肠炎的影响不同，其中口服+直肠给药疗效最好，分别提取后给药次之，混合提取后给药效果不够理想。

关键词：提取方法；给药途径；肠腑康胶囊；脾虚型溃疡性结肠炎

基金项目：陕西省科技统筹创新工程计划项目（2014KTCQ03-10）

通讯作者：张小丽，E-mail：zhangxiaoli8788@163.com，Tel：13038932330

T3-8

白虎加桂枝汤对热痹模型大鼠特征性甲基化基因表达的影响

陈 欢1，2，巨少华1，2，付文君1，2，魏江平1，2，徐世军1，2

（1.四川省中药资源系统研究与开发利用国家重点实验室培育基地，四川成都 611137；2.成都中医药大学，四川成都611137）

摘要：目的考察白虎加桂枝汤对热痹特征性甲基化基因的影响，揭示其治疗热痹的表观遗传学机制。方法采用佐剂性关节炎模型（AA）复合湿热环境（温度37℃、相对湿度70%~80%，2 h/d）制备热痹模型（HB）。第15 d后，连续灌胃给与白虎加桂枝汤（28，14和7 g·kg-1）30 d后取空白组（NG）、AA及HB大鼠膝关节滑膜，采用MeDIP-Seq测序检测滑膜DNA甲基化水平，以取NGvsHB和NGvsAA差集的方式筛选热痹特征性甲基化基因，并采用qRT-PCR检测特征性甲基化基因的表达水平。结果NGvsAA，NGvsHB及NGvsHB各有差异性甲基化基因分别为705，2418及1287个，这些甲基化差异性基因主要与炎症免疫通路，如T、B细胞受体信号通路、MAPK信号通路及抗原提呈和处理等有关，生物合成与代谢通路，如多种氨基酸代谢途径及糖类、蛋白质代谢途径等有关；热痹特征性甲基化基因共42个，与AA比较，白虎加桂枝汤高、中、低各剂量组均能抑制热痹特征性甲基化下调基因Ahcy（0.57±0.12、0.58±0.32、0.73±0.12）及Rpl3（0.65±0.24，0.65±0.24，0.62±0.30）的表达，而仅高、中剂量组促进热痹特征性甲基化上调基因Agxt（1.35±0.20，1.15±0.23）的表达水平。结论特征性基因甲基化水平变化可能是导致热痹发病的原因，白虎加桂枝汤可通过纠正特征性基因的甲基化水平达到治疗疾病的目的。

关键词：类风湿性关节炎；热痹；DNA甲基化；白虎加桂枝汤

基金项目：国家自然科学基金面上项目（81273900）；中药药理四川省青年科技创新研究团队（2014TD0007）

通讯作者：徐世军，E-mail：docxu@126.com，Tel：（028）61800231

T3-9

雷公藤内酯醇LLDT-8在抗GBM抗体诱导小鼠肾炎中的疗效作用及对Fc受体介导淋巴细胞活化的调控机制

祁 青1，2，何世君2，唐 炜2，左建平1，2

（1.上海中医药大学免疫与病毒实验室，上海 201203；2.中国科学院上海药物研究所免疫药理学研究室，上海201203）

摘要：目的研究（5R）-5-羟基雷公藤内酯醇（LLDT-8）对小鼠抗肾小球基底膜（GBM）肾炎的疗效作用，探讨LLDT-8对Fcγ受体介导的淋巴细胞活化的调控机制。方法（1）运用兔抗小鼠GBM血清诱导的雄性NZW小鼠肾炎模型，以LLDT-8口服治疗4周。观察药物对肾炎小鼠的体重、蛋白尿、肾病理变化等指标的影响作用；（2）采用流式细胞术对LLDT-8治疗干预肾炎小鼠肾浸润细胞亚群比例及表型进行分析；（3）采用免疫荧光法观察小鼠肾小球中补体沉积及炎性单核细胞浸润；（4）采用PCR法检测肾组织中Fcγ受体及炎性因子表达；（5）采用肾炎小鼠疾病高峰脾淋巴细胞，考察LLDT-8对抗血清诱导记忆性细胞应答的影响。结果（1）LLDT-8能缓解抗GBM抗体诱导肾炎小鼠的疾病进程，改善肾病变，减少肾补体沉积及免疫细胞浸润。（2）LLDT-8治疗能降低疾病高峰血清中IFN-γ及IL-6水平。（3）LLDT-8降低疾病发展各阶段的外周及肾CD11b+单核细胞Fcγ受体表达水平。（4）LLDT-8体外抑制肾炎小鼠淋巴细胞抗原特异性增殖，降低IgG及IL-6的分泌水平。结论证实了LLDT-8对于抗GBM抗体诱导的小鼠肾炎具有显著的治疗作用，阐明了LLDT-8影响单核/淋巴细胞的Fcγ受体的表达，调控抗原特异性记忆淋巴细胞应答效应并抑制肾病灶中免疫细胞浸润，从而缓解肾小球肾炎进程的疗效机制。

关键词：雷公藤内酯醇；抗GBM抗体诱导肾炎；Fcγ受体；淋巴细胞活化

基金项目：国家新药创制重大专项（012014ZX09101002-001-02）

通讯作者：左建平，E-mail：jpzuo@simm.ac.cn，Tel：（021）50806701

T3-10

DL1502新晶型对骨关节炎模型的治疗作用

王晓波1，2，赵晓悦1，王海港1，于子茹1，杨玉林1，2，孙 悦2，王淑美2，杜冠华1，2

（1.中国医学科学院北京协和医学院药物研究所，北京市药物靶点研究与新药筛选重点实验室，北京100050；2.广东药科大学，广东广州 510006）

摘要：目的观察DL1502的新晶型对大鼠膝骨关节炎所导致的关节间隙狭窄的保护作用以及对肿胀、疼痛等症状的影响，并探讨了DL1502药物新的给药方式的药效作用。方法采用关节腔药物注射法制备大鼠膝关节骨关节炎模型。对动物进行自主活动、电子压痛、步态评测等行为学评分。测量大鼠膝关节直径评判肿胀程度；应用小动物活体成像仪系统拍摄X-Ray平片测量大鼠造模侧膝关节的关节腔间隙变狭窄程度。结果与正常组相比，模型组大鼠膝关节电子压痛阈值评分显著降低（P＜0.05），而DL1502新晶型药物1.125和4.5 mg·kg-1剂量组及新给药方式组均可显著改善疼痛阈值（P＜0.05，P＜0.05和P＜0.05）。DL1502新晶型1.125和4.5 mg·kg-1剂量组以及新给药方式组可显著减轻由骨关节炎所引起的肿胀（P＜0.01，P＜0.01和P＜0.05）。X-Ray平片拍摄结果显示DL1502新晶型药物1.125和4.5 mg·kg-1剂量组能显著改善由骨关节炎导致的关节间隙狭窄程度（P＜0.01和P＜0.01），并呈剂量依赖性关系。新给药方式组关节间隙狭窄情况也有改善（P＜0.05），并且狭窄值改善情况优于DL1502新晶型1.125 mg·kg-1剂量组（P＜ 0.05），与4.5 mg·kg-1剂量组相比没有显著性差异。结论本研究采用经典的药物注射法制备大鼠膝关节骨关节炎模型，评价了DL1502新晶型对骨关节炎模型的治疗作用研究。DL1502对药物注射法所导致的大鼠膝关节关节间隙变窄有着良好的保护作用，并对骨关节炎所致的肿胀、疼痛症状有改善作用。新的给药方式同样有着显著的药效作用。

关键词：骨关节炎；新晶型；新给药方式

通讯作者：杜冠华，E-mail：dugh@imm.ac.cn，Tel：（010）63165184

T3-11

神经酰胺类鞘脂及其代谢通路参与免疫性肝纤维化发生发展的作用研究

李芳芳，刘 宁，刘 威，鲍秀琦，张 丹，孙 华

（中国医学科学院北京协和医学院药物研究所，北京100050）

摘要：目的探讨神经酰胺类鞘脂及其代谢通路在慢性免疫性肝纤维化发生发展中的作用。方法SD大鼠随机分为两组，模型组大鼠腹腔注射猪血清（PS）0.5 mL/次，每周2次，连续注射10周，诱导大鼠慢性免疫性肝纤维化，空白对照组大鼠注射等剂量生理盐水。利用血清生化指标、肝脏病理检测、超声影像检查及肝组织羟脯氨酸含量等判断肝纤维化形成及程度。HPLC-MS/MS分析血清鞘脂、特别是神经酰胺类鞘脂的含量；Western蛋白印迹法检测鞘脂代谢通路主要代谢酶的蛋白表达情况。结果SD大鼠给予PS后第4周，肝脏出现轻度纤维化。在第6，8周，肝超声检查出现纤维化造影，血清透明质酸和肝羟脯氨酸的含量较空白对照组显著升高。至第10周，可见假小叶形成。在肝纤维化发生发展过程中，血清多个鞘脂类成分，包括神经酰胺、糖基化神经酰胺含量发生明显改变。在神经酰胺的变化中，Cer（18∶1）含量在第4周显著降低，后升高但与空白对照组比较仍显著降低；Cer（22∶0），Cer（24∶1）的含量在第6周明显升高，至第8周、第10周，含量显著升高。Western蛋白印迹法检测神经酰胺代谢通路主要代谢酶的表达变化，神经酰胺合成酶2（CerS2）表达显著升高。结论神经酰胺类鞘脂可能参与了慢性免疫性肝纤维化的发生发展，CerS2可能发挥重要作用，进一步的机制研究正在进行中。

关键词：肝纤维化；神经酰胺；神经酰胺合成酶2

基金项目：国家科技计划863项目（2014AA021101）；国家自然科学基金项目（81573487）

通讯作者：孙 华，E-mail：sunhua@imm.ac.cn，Tel：（010）63165203

T3-12

构树果汁体外抗氧化及对小鼠免疫调节作用

庞素秋2，金孝勤2，王国权1

（1.华侨大学生物医学学院，福建泉州 362021；2.中国人民解放军第180医院，福建泉州 362000）

摘要：目的探讨构树果汁（BPFJ）体外抗氧化作用及对小鼠的免疫活性影响。方法从清除超氧阴离子自由基（O2-·）、羟基自由基（·OH）、DPPH自由基等方法观察了BPFJ体外抗氧化作用；用刀豆蛋白（ConA）诱导小鼠脾淋巴T细胞增殖细胞模型，MTT法测定细胞增殖并检测BPFJ对小鼠脾T淋巴细胞增殖的影响。结果BPFJ能显著清除O2-·、·OH和DPPH自由基，在0.1～2.0 g·L-1范围内呈现剂量效应关系。小鼠T淋巴细胞在ConA刺激下48 h后，MTT吸收度显著高于正常对照组。而BPFJ组与ConA组比较则显著降低了MTT吸收度。结论BPFJ有较强的体外抗氧化作用并能显著提高小鼠的免疫作用。BPFJ是一种抗氧化功效较强的活性物质，具有很好的保健功能。

关键词：构树果汁；抗氧化；自由基；免疫调节

基金项目：福建省泉州市科技项目（200712Z）

通讯作者：王国权，E-mail：wangguoqun@hqu.edu.cn

T3-13

清除衰老细胞治疗小鼠放射性肺纤维化机制

潘 金1，徐燕锋1，张俊伶2，李德冠2，王月英2，陈孟毅1，刘 冰1，周道洪3，孟爱民1

（1.中国医学科学院协和医学院，医学实验动物研究所，北京协和医学院比较医学中心，北京 100221；2.中国医学科学院放射医学研究所，天津市放射医学分子核医学重点实验室，天津 300129；3.Department of Pharmaceutical Sci⁃ences，Winthrop P.Rockefeller Cancer Institute，Univer⁃sity of Arkansas for Medical Sciences，Little Rock，Arkansas，USA，72205）

摘要：目的放射性肺纤维化是放射性肺损伤的晚期改变，发病机制复杂，临床上缺乏有效的治疗方法，患者预后差。对放射性肺纤维化机制及干预的研究是当前的研究热点。ABT-263是抗肿瘤新药，近期发现具有清除衰老细胞、改善造血干细胞辐射损伤的作用。研究在前期研究的基础上建立局部照射诱导的肺纤维化小鼠模型，观察ABT-263对放射性肺纤维化的治疗作用，探讨其作用机制。方法健康雄性C57BL/6J小鼠分为对照组（Control组）、给药组（ABT组）、照射组（IR组）和治疗组（IR+ABT组）。小鼠接受17Gy右侧胸部X射线照射，照射后第16周开始灌胃给予ABT-263 50 mg·kg-1，每疗程给药5 d，停药2周，给药2个疗程后进行观察。两个疗程治疗后CT检查小鼠肺实质的损伤情况。病理观察小鼠肺组织结构的变化、胶原生成（Masson检测）和β-半乳糖苷酶（β-Gal）表达。免疫荧光检测肺组织中肺表面活性前体蛋白C（Pro-SPC）和P16的表达。基因PCR芯片技术检测肺组织纤维化相关基因的表达。结果胸部CT结果显示ABT治疗可以降低放射引起的小鼠肺组织的HU值升高（P＜0.05）；降低照射引起小鼠肺脏指数显著升高（P＜0.05）；减轻受照小鼠肺组织病理变化评分（P＜0.01），减少胶原沉积面积（P＜0.01），降低β-Gal表达细胞比例（P＜0.01）；可以抑制受照小鼠Ⅱ型肺泡细胞P16表达（P＜0.01）；纤维化相关基因芯片测定结果显示ABT可以降低受照小鼠肺组织中Bcl-2、Col3a1、Col1a2、Ⅱ1α、Il1β mRNA显著升高（P＜0.05或P＜0.01）。结论上述研究结果显示ABT263可以减轻小鼠放射性肺纤维化的损伤性改变，降低衰老Ⅱ型肺上皮细胞，抑制Bcl-2及肺组织炎症性因子及纤维化相关因子的表达。研究结果将为放射性肺纤维化防治提供新的思路。

关键词：放射性肺损伤；肺纤维化；细胞衰老；组织干细胞；Ⅱ型肺泡上皮细胞

基金项目：国家自然科学基金（81573094；81372928）

通讯作者：孟爱民，E-mail：ai_min_meng@126.com，Tel：13911809579

T3-14

异甘草酸镁对小鼠放射性肺纤维化的改善作用及其机制的初步探讨

张 攀，刘 涛，王芳芹，阳群芳，陈晓红

（第三军医大学药学系药理教研室，重庆 400038）

摘要：目的探讨异甘草酸镁（MgIG）对放射性肺纤维化的作用及其抗纤维化的机制。方法将SPF级雌性C57小鼠随机分为正常对照组、辐照模型组、异甘草酸镁对照组、异甘草酸镁治疗组和地塞米松治疗组，每组10只。除正常对照组和异甘草酸镁对照组，其余各组小鼠均给予单次15 Gy60Coγ射线全肺照射，照射后12周处死小鼠取左肺下叶组织，免疫组化法观察肺组织Ⅰ型、Ⅲ型胶原蛋白及转化生长因子-β1（TGF-β1）信号通路蛋白表达变化。结果各组小鼠肺组织均有Ⅰ型、Ⅲ型胶原及TGF-β1，Psmad2，Psmad3不同程度的表达，模型组明显高于正常对照组，异甘草酸镁治疗组与地塞米松治疗组相较于模型组各蛋白表达均有下降。结论异甘草酸镁可通过抑制小鼠肺组织TGF-β1信号通路改善放射性肺纤维。

关键词：放射性肺纤维化；异甘草酸镁；转化生长因子β1

基金项目：国家自然科学基金项目（81272995）

通讯作者：陈晓红，E-mail：pharma821@163.com，Tel：13228683975

T3-16

Nrf2 inhibits epithelial-mesenchymal transition by suppressingsnailexpressionduringpulmonary fibrosis

ZHOU Wen-cheng1，MO Xiao-ting1，ZHANG Zhi-hui2，CUI Wen-hui1，GAO Jian3

（1.School of Pharmacy，Anhui Medical University，Hefei 230032，China；2.The First Affiliated Hospital of Anhui Medical University，Hefei 230022，China；3.The Second Hospital of Dalian Medical University，Dalian 116023，China）

Abstract：OBJECTIVEEpithelial-mesenchymal transition（EMT）is a phenotype conversion that plays a critical role in the development of pulmonary fibrosis（PF）.It is known that a transcription factor snail could regulate the progression of EMT.Nuclear factor erythroid 2 related factor 2（Nrf2），a key regulator of antioxidant defense system，protects cells and tissues against oxidative stress.However，it is not known whether Nrf2 regulates snail thereby modulating the development of PF.MEHODSBleomycin（BLM）was intratracheally injected into both Nrf2-knockout（Nrf2-/-）and wild-type mice to compare the development of PF.Rat type II alveolar epithelial cells（AECs）RLE-6TN were treated with a specific Nrf2 activator sulforaphane，or transfected with Nrf2 and snail siRNAs to determine their effects on transforming growth factor β1（TGF-β1）-induced EMT.RESULTSBLM-in⁃duced EMT and lung fibrosis were more severe in Nrf2-/-mice compared to wild-type mice.In vitro，sulforaphane treatment attenuated TGF-β1-induced EMT，accompa⁃nied by the down-regulation of snail.Inversely，silencing Nrf2 by siRNA enhanced TGF-β1-induced EMT along with the expression of snail.Interestingly，silencing snail by siRNA reduced TGF-β1-induced EMT even in the pres⁃ence of sulforaphane in RLE-6TN cells.CONCLUSIONThese findings suggested that Nrf2 may attenuate EMT and fibrosis process through regulating the expression of snail in PF.

Key words：nuclear factor erythroid 2-related factor2；snail；epithelial-mesenchymaltransition；pulmonaryfibrosis

Foundation item：The project supported by National NaturalScience Foundation ofChina（81274172，81473267，30801535，and 81470003）

Corresponding author：GAO Jian，E-mail：gaojianay⁃fy@163.com

T3-17

Anti-inflammatory effect of extracts ofDendropanax dentigeandLycopodiastrum casuarinoideson rheu⁃matoid arthritis and their underlying mechanism in rats

CHEN Gang，LIU Zhi-kai，HAN Chen-yun，LI Jia-shuang，ZOU Yi-ping

（Key Laboratory of Natural Active Pharmaceutical Con⁃stituents of Jiangxi Province，Yichun University，Yichun 336000，China）

Abstract：OBJECTIVETo investigate the anit-inflamma⁃tory effect of extracts ofDendropanax dentiger（Harms）Merr andLycopodiastrum casuarinoides（Spring）Holub on rheumatoid arthritis（RA）using adjuvant arthritis（AA）rat model and possible mechanisms.METHODSThe AA rat model of RA was induced in adult Sparague-Dawley（SD）rats by injecting of the adjuvant at base oftail.One-week-old male SD rats were randomly divided into the following groups：normal saline group（blank control），D.dentigerdecoction group（80g·kg-1·d-1），L.Casuarinoidesdecoction group（80 g·kg·d-1），the total of glucoside Tripterygium（GTT）group（positive control，2 mg·kg-1·d-1）.They were administered orally for 6 weeks.Histopathology of tissues arthritis rats was ob⁃served by H.E staining.The volume of paw swelling was measured and the arthritis inflammation index was calcu⁃lated.The expressions of tumor necrosis factor-α（TNF-α）and interlukin-1β（IL-1β）were detected by the ELISA assay.In addition，previous study has reported that plant-derived miRNAs play a role for cross kingdom regu⁃latory potential.Thus，we also performed RNA-seq tech⁃nique to identify bioactive miRNAs via comparative tran⁃scriptome analysis betweenD.dentigerandL.Casuari⁃noides.RESULTSComparing with AA model group，the volume of paw swelling and the arthritis index were increased significantly in the AA rat model group（P＜0.01），suggesting that the AA model rats were prepared properly.Compared with the AA model group，the vol⁃ume of paw swelling ofD.dentigerdecoction group，L. Casuarinoidesdecoction group was decreased by 25.2% and 10.3%，respectively，and the arthritis index was de⁃creased by 27.2%and 18.3%，respectively.Compared with AA model group，TNF-α protein expression ofD. dentigerdecoction group andL.Casuarinoidesdecoction groups were decreased by 16.3%and 14.7%，and IL-1β protein expression was decreased by 23.6%，18.9%（P＜0.05，P＜0.01），respectively.Besides，we found that some plant-derived homologous miRNAs（such as miRNA192 and miRNA30a）associated with cell apopto⁃sis processing have been screened out via comparative transcriptome analysis.But the underlying mechanisms about two miRNAs function needs much more investi⁃gate.CONCLUSIONResults showed significant anti-in⁃flammatory effect of aqueous extracts ofD.dentigerandL.Cauarinoidesand justifying their therapeutic role in in⁃flammatory condition.Furthermore，anti-inflammatory ef⁃fect ofD.dentigerandL.Cauarinoidesmay be attribute to theherb-derivedmiRNAs cross-kingdom regulation.

Key words：rheumatoid arthrits；aqueous extracts ofD. dentigerandL.Cauarinoides；anti-inflammatory effect；miRNA；cross-kingdom regulation

Foundation item：The project supported by Science and Technology External Cooperation Key Foundation of Jiangxi Province（20151BDH80020）

Corresponding author：ZOU Yi-ping，E-mail：zyping66@ 163.com

T3-18

Increased BAFF in serum is a novel biomarker related to disease activity in rheumatoid arthritis

ZHANG Ling-ling1，XIAO Hui2，Zhang Feng1，WU Yu-jing1，SHU Jin-ling Shu1，LI Ying1，TAI Yu1，WEI Wei1

（1.Key Laboratory of Antiinflammatory and Immunophar⁃macology of Education Ministry，Institute of Clinical Pharmacology，Anhui Medical University，Hefei，China；2.Department of Rheumatology and Immunology，No.1 AffiliatedHospital，AnhuiMedicalUniversity，Hefei，China）

Abstract：OBJECTIVETo investigate the contribution of B cell activating factor of TNF family（BAFF）in the prolif⁃eration and activation of B cell in rheumatoid arthritis（RA），and to clarify whether high levels of BAFF is asso⁃ciated with clinical variables and lab parameters.METH⁃ODSBlood samples and peripheral blood mononuclear cells from RA patients and healthy controls（HCs）were collected and isolated respectively.Clinical variables and lab parameters，BAFF level，cytokines and immunoglob⁃ulins in serum were evaluated at entry.B cell subpopula⁃tions，BAFF receptors（BAFFR，BCMA，TACI），and al⁃ternative and canonical NF-κB pathway in B cell were an⁃alyzedin vivoandin vitro.RESULTSIn RA patients，BAFF level and activated B cell subsets increased signifi⁃cantly.BAFF level was associated with CRP，RF，DAS28，swollen joint counts and tender joint counts. BAFFR，BCMA，TACI on B cells in RA over expressed. The expressions of MKK3，MKK6，p-p38，p-NF-κB65，TRAF2，NF-κB52 were higher than that in HCs.In vitro，BAFF up regulated activated B cell subset and the ex⁃pressions of BAFFR，BCMA and TACI.BAFF also en⁃hanced the expressions of MKK3，MKK6，p-p38，p-NF-κB65，TRAF2，NF-κB52.CONCLUSIONIncreased BAFF in serum is associated with the disease activity of RA，BAFF involves in the proliferation and activation of B cell in RA through alternative and canonical NF-κB pathway，indicating that BAFF might be a novel biomark⁃er of diagnosis and therapy.

Key words：rheumatoid arthritis；B cell；BAFF；disease activity

Foundation item：The project supported by National NaturalScience Foundation ofChina（81473223，31100640，81302517 and 81330081）；and China Post⁃doctoral Science Foundation（2013M540509）

Corresponding aucthor：WEI Wei，E-mail：wwei@ahmu. edu.cn

T3-19

Matrine ameliorates experimental autoimmune encepha⁃lomyelitis by regulating the differentiation of encepha⁃litogenic Th1/Th17 and Th2/regulatory T cells

ZHANG Ming-liang1*，KAN Quan-cheng2*，ZHANG Hui-jun1，ZHU Lin1

（1.Department of Pharmacy，The First Affiliated Hospital of Zhengzhou University，Zhengzhou 450052，China；2.The First Affiliated Hospital of Zhengzhou University，Zhengzhou 450052，China）

Abstract：OBJECTIVEExperimental autoimmune en⁃cephalomyelitis（EAE），the classical animal model for multiple sclerosis（MS）is triggered by an impaired bal⁃ance of T helper（Th）cells and regulatory T（Tregs）cells.Matrine（MAT），a quinolizidine alkaloid derived from the herb Radix Sophorae Flave，has been shown to ame⁃liorate the clinical signs，inflammatory infiltration，demye⁃lination in acute EAE rats.However，whether MAT pro⁃tect from EAE by adjusting Th and Treg cells response in specific-cellular and molecular level is unknown.METH⁃ODSHerein，MAT was tested for its effects on Th1，Th2，Th17 and Treg cells in the spinal cord of EAE mice and splenocyte-extracted from EAE mice with MOG35-55-restimulated，respectively.RESULTSOur findings re⁃vealed that MAT significantly inhibit the proliferation of splenocyte，and remarkably down-regulate the differenti⁃ation of Th1/Th17 cells with decreased expressions of CD4+IFN-γ+cells and CD4+IL-17+cellsin vivoand IL-17，IFN-γ，ROR-γt，T-betin vitro，meanwhile it dramatically up-regulate the Th2/Treg cells response associated with increased levels of CD4+TGF-β1+cells and CD4+IL-10+cellsin vivoand IL-4，IL-10，TGF-β1，Foxp3 and GATA3in vitro.CONCLUSIONConsidering the effective thera⁃peutic effects of MAT on EAE，it′s worth to find its new values on other autoimmune diseases.

Key words：matrine；experimental autoimmune encepha⁃lomyelitis；T helper cells；regulatory T cells

Foundation item：The project supported by National Natural Science Foundation of China（31570357）

Corresponding author：ZHU Lin，E-mail：zhulin66zhulin@ 126.com，Tel：（0371）66913380

*Co-first author.

T3-20

Mechanism of TSCS promote anti-Fas antibody-in⁃duced FLS apoptosis via blocking MC-tryptase-PAR-2-Rho-FLS signaling pathway

LI Shi-gang1，2，LIU Wei2，YU Ling-ling2，JIA Yun-li2，ZHENG Qian-qian2，CHEN Xian-yong2

（1.Renhe Hospital，2.Medical College，China Three Gorges University，Yichang 443002，China）

Abstract：OBJECTIVEIt has been supposed that mast cells have important participation in the physiopathology of RA，however，the role of mast cells in the pathogene⁃sis of RA remains unclear.In this study，we observed the antiapoptotic effects of tryptase released by mast cell on RA synovial fibroblasts.METHODSMast cells and fi⁃broblasts synovial were obtained from mouse.Chemical mediator release was assessed by measuring β-hexosa⁃ minidase release.TSCS and bone marrow-derived mast cells were co-cultured；the toxic effects of TSCS on mast cell was measured by MTT and CCK-8 method；the releasing amount of tryptase in cell supernatants was measured by Elisa assay；the expression of FLS cell membrane receptor PAR-2 was detected by flow cytome⁃try；the apoptosis of FLS cell was detected through flow cytometry and Western blotting；the level of activated Rho-GTP was detected by the pull-down method and Western blotting.RESULTSIn this study，we observed the antiapoptotic effects of tryptase released by mast cell on RA synovial fibroblasts，and found that tryptase signif⁃icantly increased the expression of PAR-2 on the surface of fibroblast-like synovial cell，significantly activated Rho kinase，significantly inhibited apoptosis of fibroblast-like synovial cell induced by CH11.The release rates of βhexosaminidase and the level of tryptase from bone mar⁃row-derived mast cells after stimulation with different anti⁃gen and co-cultured with TSCS significantly decreased compared to the control group.In the co-culture system of mast cells and fibroblast-like synovial cells，TSCS treatment significantly inhibited Rho kinase（P＜0.05），significantly promoted apoptosis of fibroblast-like synovi⁃al cell induced by CH11（P＜0.05）.CONCLUSIONThese results demonstrate that tryptase may play a key role in the physiopathology of RA.TSCS can inhibit mast cells activation and promote FLS cells apoptosis.This provide theoretical and experimental basis for the study of mast cells as targets for new anti-RA drugs.

Key words：total saponins ofChaenomeles speciosa；FLS；apoptosis；mast cells；cytokines

Foundation item：The project supported by National NaturalScience Foundation ofChina（81274166，81673665）；Project in Hubei Province Department of Ed⁃ucation（B20101201）；Yichang City Technology Bureau Project（2010A01301-04）；and China Three Gorges Uni⁃versity Research Fund（0620080702）

Corresponding author：LI Shi-gang，E-mail：fox201@ 163.com

T3-21

The GSK3b sensing metabolism controls NLRP3 inflammasome activation

HAN Sheng-na1，2，ZHANG Li-rong2，Wajahat Z MEHAL1，OUYANG Xin-shou1

（1.Section of Digestive Diseases，Yale University，New Haven，CT，USA，06520；2.Department of Pharmacology，Basic Medical College，Zhengzhou University，Zhengzhou 450001，China）

Abstract：OBJECTIVETo identify the role of GSK3 iso⁃form inhibition on inflammasome activation.METHODSThe NLRP3 inflammasome was activated by typical LPS/ATP and host-derived metabolites in primary mouse mac⁃rophages.The pharmacological inhibition of GSK3 iso⁃forms on inflammasome activation was assayed by quan⁃tifying IL-1β in the supernatant，and activated caspase-1 in cell lysates using highly selective inhibitors.Further molecular mechanisms were investigated by protein pulldown assay，confocal imaging using forced gene expres⁃sion system and endogenous protein tagged mouse mac⁃rophages.RESULTSPharmacologicalinhibition of GSK3-β，but not GSK3-α isoform suppressed NLRP3 in⁃flammasome activation in response to ATP，urate crystal and the microbial alkaloid toxin staurosporine.GSK3-β in⁃hibition did not inhibit melanoma 2（AIM2）inflamma⁃some activation in response to double-stranded DNA（ds⁃DNA）and did not affect non-canonical caspase-11 in⁃flammasome activation.GSK3-β inhibition suppressed high glucose mediated NLRP3 inflammasome activation. Mechanistically，GSK3-β inhibition blocked NLRP3 in⁃flammasome by preventing proIL-1β transcription，reduc⁃ing caspase-1 activation and ASC speck formation. GSK3-β inhibition blocked NLRP3 inflammasome activa⁃tion without affecting the level of reactive oxygen species（ROS）which is a crucial component in initialing inflam⁃masome activation.Further studies revealed that GSK3-β directly binds to ASC by both co-forced expression and endogenous protein level.Interestingly，we found ASC can be glycosylated in response to inflammasome activa⁃tion，and GSK3-β inhibition reduced ASC glycosylation. Consistently，the O-GlcNAc transferase（OGT）deficient mouse macrophages showed the significant reduction of mature IL-1β secretion in response to NLRP3 inflamma⁃some activation.CONCLUSIONOur results demonstrate a critical role of metabolism-sensing GSK3-β pathway in mediating NLRP3 inflammasome activation，thus defin⁃ing a new therapeutic target for sterile inflammation.

Key words：GSK3-β；NLRP3 inflammasome；IL-1β secretion；O-GlcNAc transferase；glycosylation

Corresponding author：OUYANG Xin-shou，E-mail：xinshou.ouyang@yale.edu

T3-22

Curcuma′s extraction attenuates propranolol-induced psoriasis like in mice by inhibition of keratin，prolif⁃erating cell nuclear antigen and Toll-like receptor expression

LI Jin-qi1，2，3，ZHANG Shu-han2，TONG Rong-sheng1，2，3，HE Dan2，ZHONG Zhen-dong1，SHE Shu-ya1*

（1.Sichuan Academy of Medical Sciences&Sichuan Provincial People′s Hospital，Chengdu 610000，China；2.School of Medicine，University of Electronic Science and Technology of China，Chengdu 610000，China；3.Sichuan Key Laboratory for Individualized Drug Therapy，Chengdu 610000，China）

Abstract：OBJECTIVERecently，some reports show that curcuma has a good curative effect on the treatment of psoriasis；while curcuma has many components，so the present study was designed to investigate the effec⁃tive parts of curcuma on treatment of psoriasis，in addi⁃tion，we will explore the mechanisms of curcuma thera⁃peutic effect.METHODSFirst，we observed that curcu⁃ma′s extractions effect on mitosis of mouse vaginal epi⁃thelial cells；then psoriasis like was induced by wiping im⁃iquimod in the surface of the binaural dorsal skin of the animal，the score of skin damage was measured on days 7 and 14；in order to explore the mechanisms of curcuma’s extractions on psoriasis，immune factors ex⁃pression（CK14，CK16，CK17，PCNA，TLR-2，TLR-4，TLR-9）was determined in propranolol induced psoriasis like.RESULTSCurcuma′s extraction prohibited the mito⁃sis of mouse vaginal epithelial cells；curcuma’s extrac⁃tions produced a significant and dose dependent inhibi⁃tion of psoriasis during the entire duration of the study in imiquimod induced psoriasis like；and the expression of immune factors（CK14，CK16，CK17，PCNA，TLR-2，TLR-4，TLR-9）was also found to be less in the curcuma′s extraction treated group as compared to control.CON⁃CLUSIONWe believe that the curative effectof curcuma’sextraction may due to its inhibition effect on the expres⁃sion of immune factors.Our results contribute towards validation of curcuma in the treatment of psoriasis and other joint disorders.

Key words：psoriasis；curcuma′s extract；ionlmmune factors

T3-23

Salvianolic acid A alleviates renal injury in systemic lupus erythematosus induced by pristane in BALB/c mice

LIN Yi-huang1，YAN Yu1，ZHANG Hui-fang1，CHEN Yu-cai1，HE Yang-yang2，WANG Shou-bao2，FANG Lian-hua1，LYU Yang3，DU Guan-hua2

（1.Beijing Key Laboratory of Drug Targets Identification and Drug Screening，3.Beijing Key Laboratory of Poly⁃morphic Drugs，Institute of Materia Medica，Chinese Academy of Medical Sciences and Peking Union Medical College，Beijing 100050，China；2.State Key Laboratory of Cardiovascular Disease，Fuwai Hospital，National Center for Cardiovascular Diseases，Beijing 100037，China）

Abstract：OBJECTIVETo investigate the effects of sal⁃vianolic acid A（SAA）in systemic lupus erythematosus（SLE）induced by pristane in BALB/c mice，this study was performed.METHODSLupus mice were estab⁃lished by confirming elevated levels of autoantibodies and IL-6 after intraperitoneal injection of pristane.Micewere then treated with daily oral doses of SAA for 5 months in parallel with mice treated with prednisone and aspirin as positive controls.The levels of autoantibodies were monitored at monthly intervals and nephritic symp⁃toms observed by hematoxylin and eosin（H&E）and pe⁃riodic acid-Schiff（PAS）staining.Western blot analysis of renal tissue was also employed.RESULTSSAA treatment caused a significant reduction in the levels of anti-Sm autoantibodies and reduced renal histopatho⁃logical changes and pathological effects.SAA treatment also significantly inhibited the phosphorylation of IKK，IκB and NFκB in renal tissues of lupus mice.CONCLU⁃SIONThe results suggest that SAA alleviates renal inju⁃ry in pristane-induced SLE in BALB/c mice through inhibi⁃tion of phosphorylation of IKK，IκB and NFκB.

Key words：salvianolic acid A；systemic lupus erythema⁃tosus；renal injury；autoantibodies；pristane；NF-κB

Foundation item：The project supported by National NaturalScience Foundation ofChina（81573645，81673422）

Corresponding authors：FANG Lian-hua，E-mail：fanglh@ imm.ac.cm；DU Guan-hua，E-mail：dugh@imm.ac.cn.

T3-15

Interaction of Wnt/β-catenin and Nrf2 pathways in cig⁃arette smoke-induced inflammation and emphysemaCUI Wen-hui1，MO Xiao-ting1，ZHOU Wen-cheng1，ZHANG Zhi-hui2，GAO Jian3

（1.School of Pharmacy，Anhui Medical University，Hefei 230032，China；2.The First Affiliated Hospital of Anhui Medical University，Hefei 230022，China；3.The Second Affiliated Hospital of Dalian Medical University，Dalian 116023，China）

OBJECTIVEThe present study aimed to in⁃vestigate the relationship between Wnt/β-catenin and Nrf2 signaling pathways，and understanding the mecha⁃nisms underlying the process of inflammatory in chronic obstructive pulmonary disease（COPD），which was a se⁃rious disease of respiratory system.METHODSWe du⁃plicate the emphysema model with porcine pancreatic elastase（PPE）in Nrf2-/-and WT mouse for 21d，and in⁃traperitoneal injection of LiCl，the activator of Wnt/β-catenin signaling pathway from 14 d to the end.Hematox⁃ylin and eosin（H&E）staining was performed to assess the histopathologiclevel，and immunohistochemistry（IHC）for Mac-3（the marker of macrophagocyte）and Ly6G（the marker of neutrophil）was used to observe the inflammatory infiltrate，while the levels of Wnt/βcatenin and Nrf2 signaling pathways related proteins heme oxygenase-1（HO-1），NAD（P）H：quinone oxido⁃reductase 1（NQO1），and the expression of inflammatory cytokine interleukin-6（IL-6）were detected by Western blotting of lung tissues.In vitro，cigarette smoke extract（CSE）-treated normal human bronchial epithelial（NHBE）cells，cell viability was examined by MTT assay，and then we treated recombinant human Wnt3a，siNrf2 and siWnt3atomeasure theexpressionofWnt3a，βcatenin，Nrf2，HO-1，NQO-1，and IL-6.Cellular immuno⁃fluorescence staining was employed to identify the nucle⁃ar translocation of Nrf2.RESULTSWe found that the LiCl-treated group has markedly decreased the damage of alveolar structure and inflammatory signs than the model group of WT mice rather than Nrf2-/-group.It also seen that LiCl not only increased β-catenin，but it also led to a comparable increase in Nrf2，HO-1，NQO1，and decrease of IL-6 compared with WT model groups but ex⁃cept to Nrf2-/-groupin vivo.And it showed that Wnt3atreatment has significantly increased the nuclear translo⁃cation of Nrf2 and the expression of HO-1 and NQO1，re⁃duced the IL-6 release，while there has no significance when Nrf2 was blocked in CSE-induced NHBE cells.CONCLUSIONOur results demonstrated that Wnt3a/βcatenin significantly balanced oxidative stress and attenu⁃ated inflammation reaction by promoting Nrf2 nuclear translocation and activity.

chronic obstructive pulmonary disease；em⁃physema；interleukin-6；inflammation；nuclear factor erythroid-2-related factor-2；Wnt/β-catenin

GAO Jian，E-mail：gaojianayfy@ 163.com

吉林省科技发展计划项目（20140204060YY）

洪 铁，E-mail：hongtie@jlu.edu.cn，Tel：13843053169

Foundation item：The project supported by National NaturalScience Foundation ofChina（81274172，81473267，30801535，81470003）