專題3:抗炎癥和免疫藥理學(xué)

T3-1

人參地黃桑葚片對小鼠免疫功能的影響

付 萌，洪鐵

（吉林大學(xué)藥學(xué)院藥理教研室，吉林長春 130021）

目的探索人參地黃桑葚片對小鼠免疫功能的影響。方法將三批雄性ICR小鼠每批隨機分為對照組、人參地黃桑葚片高劑量組（2.250 g·kg-1）、人參地黃桑葚片中劑量組（0.750 g·kg-1）、人參地黃桑葚片低劑量組（0.375 g·kg-1），每組15只，連續(xù)灌胃給藥1個月后，稱量并計算小鼠免疫器官指數(shù)（胸腺和脾臟），采用綿羊紅細胞（SRBC）誘導(dǎo)法測定小鼠遲發(fā)型變態(tài)反應(yīng)（DTH）；采用乳酸脫氫酶（LDH）法測定自然殺傷（NK）細胞的活性、MTT法測定刀豆蛋白A（ConA）誘導(dǎo)的脾淋巴細胞增殖能力；采用滴片法測定腹腔巨噬細胞對雞紅細胞（CRBC）的吞噬作用。結(jié)果人參地黃桑葚片高、中、低劑量組均能顯著增加小鼠胸腺指數(shù)及脾指數(shù)（P＜0.05）；人參地黃桑葚片高、中劑量組能顯著增強小鼠NK細胞活性及SRBC誘導(dǎo)的小鼠DTH（P＜0.05）；人參地黃桑葚片高劑量組能顯著提高ConA誘導(dǎo)的脾淋巴細胞的增殖能力及腹腔巨噬細胞對CRBC的吞噬作用（P＜0.05）。結(jié)論人參地黃桑葚片能夠顯著增強小鼠的免疫功能。

人參地黃桑葚片；免疫功能

T3-2

利用斑馬魚模型研究姜黃素抗炎的分子機制

王榮春，韓利文，何秋霞，陳錫強，劉可春

（山東省科學(xué)院生物研究所，山東省生物傳感器重點實驗室，山東省科學(xué)院藥物篩選技術(shù)重點實驗室，山東省生物檢測技術(shù)工程實驗室，山東濟南 250014）

摘要：目的利用斑馬魚炎癥模型探討姜黃素抗炎作用，分析姜黃素抗炎的分子機制。方法采用3dpf（days after fertil?ization）轉(zhuǎn)基因斑馬魚Tg（Lyz：GFP）幼魚，用姜黃素0，5，10，30和50 μmol·L-1處理1 h后，通過用硫酸銅20 μmol·L-1處理造成急性炎癥，體式顯微鏡下觀察姜黃素對中性粒細胞的遷移、定位，并統(tǒng)計遷移中性粒細胞的數(shù)量；通過脂多糖（LPS）10 μmol·L-1處理受精后24 h野生型斑馬魚AB品系造成炎癥模型，并姜黃素0，5，10，30和50 μmol·L-1，通過NO和ROS檢測試劑盒檢測NO和ROS的表達水平，通過熒光倒置顯微鏡進行拍照，統(tǒng)計熒光強度。結(jié)果在硫酸銅造成的轉(zhuǎn)基因斑馬魚急性炎癥模型下，與對照處理相比，姜黃素30和50 μmol·L-1抑制硫酸銅造成的中心粒細胞向斑馬魚測線的滲漏，滲漏的中性粒細胞數(shù)目明顯減少（P＜0.05）；LPS和姜黃素處理斑馬魚幼魚，姜黃素30和50 μmol·L-1可以降低LPS處理誘導(dǎo)的NO和ROS的表達水平（P＜0.05）。結(jié)論本研究通過不同斑馬魚炎癥模型驗證了姜黃素具有抗炎的作用，其機制是通過抑制NO和ROS信號的釋放，減少中性粒細胞的滲漏，從而緩解炎癥。

關(guān)鍵詞：姜黃素；斑馬魚；炎癥；硫酸銅；脂多糖；

基金項目：國家自然科學(xué)基金青年基金項目（31400979）；山東省科學(xué)院青年基金項目（2015QN010）

通訊作者：劉可春，E-mail：hliukch@sdas.org，Tel：（0531）82605352

T3-3

雙環(huán)醇抗炎作用機制研究

劉 寧，李芳芳，張 丹，鮑秀琦，孫 華

（中國醫(yī)學(xué)科學(xué)院/北京協(xié)和醫(yī)學(xué)院藥物研究所，北京100050）

摘要：目的研究雙環(huán)醇抗炎機制及可能靶點。方法佛波酯（PMA）誘導(dǎo)分化THP-1單核細胞成巨噬細胞，LPS刺激。ELISA檢測細胞上清中炎癥因子含量，Western蛋白印跡法檢測蛋白表達。siRNA進行基因沉默，進一步分析炎癥因子及蛋白表達的變化。3T3-L1前脂肪細胞分化為脂肪細胞，油紅O染色。結(jié)果雙環(huán)醇對LPS刺激的人巨噬細胞炎癥因子TNF-α、IL-6、IL-1β的釋放表現(xiàn)出顯著的抑制活性，其能抑制PI3K的表達，促進IκB表達，抑制NF-κB入核，但對Akt、IKK磷酸化無明顯影響。進一步的研究顯示，雙環(huán)醇顯著促進PPARγ表達，沉默PPARγ基因，雙環(huán)醇對IκB的促表達作用減弱，抑制炎癥因子分泌的作用降低。油紅O染色結(jié)果顯示，雙環(huán)醇可促進前脂肪細胞的分化，但作用弱于羅格列酮。結(jié)論雙環(huán)醇具有良好的抗炎活性，其抗炎作用與促進PPARγ表達，進而促進IκB表達，抑制NF-κB入核有關(guān)。

關(guān)鍵詞：雙環(huán)醇；LPS；細胞因子；PPARγ；IκB；

基金項目：國家科技計劃863項目（2014AA021101）；國家自然科學(xué)基金項目（81173073）

通訊作者：孫 華，E-mail：sunhua@imm.ac.cn，Tel：13811929788

T3-4

蝦青素對博來霉素致大鼠肺纖維化的干預(yù)作用及機制

王芳芹，劉 濤，張 攀，陽群芳，陳曉紅

（第三軍醫(yī)大學(xué)藥學(xué)系藥理教研室，重慶 400038）

摘要：目的探討蝦青素對博萊霉素所致SD大鼠肺纖維化的干預(yù)作用。方法將SD大鼠隨機分為假手術(shù)組、蝦青素組、博來霉素組、蝦青素+博來霉素組。經(jīng)氣道注入博萊霉素（5 mg·kg-1）或生理鹽水后，每天灌服蝦青素（2 mg·kg-1）或植物油連續(xù)14 d，最后一次給藥24 h后取材。計算肺系數(shù)，進行HE染色及Masson染色，顯微鏡下觀察大鼠肺部炎癥及纖維化程度；酶聯(lián)免疫夾心法檢測血清中TGF-β1水平；免疫組化觀察肺間質(zhì)α-SMA蛋白表達變化；堿水解法檢測羥脯氨酸含量；Western蛋白印跡法法檢測ERK1/2及Smad2/3蛋白磷酸化水平。結(jié)果與假手術(shù)組相比，博來霉素組肺損傷嚴重，有明顯的肺纖維化病灶形成；與模型組比較，藥物治療組肺損傷明顯減輕，膠原蛋白沉積減少，羥脯氨酸含量也明顯降低（P＜0.01），α-SMA蛋白表達減少，TGF-β1水平明顯降低（P＜0.05），同時ERK1/2蛋白及Smad2/3蛋白磷酸化水平明顯下降。結(jié)論蝦青素能明顯改善肺纖維化，其作用機制可能與降低TGF-β1水平，抑制ERK1/2蛋白及Smad2/3蛋白磷酸化有關(guān)。

關(guān)鍵詞：蝦青素；肺纖維化；博來霉素

基金項目：國家自然科學(xué)基金項目（81472912）

通訊作者：陳曉紅，E-mail：pharsunr@126.com，Tel：（023）68752364

T3-5

炎癥免疫反應(yīng)軟調(diào)節(jié)—藥物研究新方向

魏偉

（安徽醫(yī)科大學(xué)臨床藥理研究所，抗炎免疫藥物教育部重點實驗室，抗炎免疫藥物安徽省協(xié)同創(chuàng)新中心，安徽合肥230032）

摘要：炎癥免疫反應(yīng)（IIR）是機體炎癥免疫相關(guān)組織、器官和細胞依據(jù)內(nèi)外環(huán)境變化所表現(xiàn)出的適度或異常的系統(tǒng)反應(yīng)，幾乎涉及機體大多系統(tǒng)的保護或病理反應(yīng)。參與IIR的細胞不僅涉及常見的炎癥免疫細胞（如巨噬細胞、樹突細胞、T細胞、B細胞等及其亞型），而且涉及非傳統(tǒng)的炎癥免疫細胞（如膠質(zhì)細胞、內(nèi)皮細胞、上皮細胞、成纖維細胞、滑膜細胞、肝細胞等），它們有著類似的或共性的細胞信號轉(zhuǎn)導(dǎo)。目前臨床使用的影響IIR藥物雖然在治療中發(fā)揮了較好療效，但出現(xiàn)的不良反應(yīng)尤其是肝、腎、胃腸、惡性感染、誘發(fā)腫瘤等嚴重毒性令人困惑，這些藥物通過不同機制在抑制IIR所致病理反應(yīng)的同時，也損害了組織、器官和細胞的生理功能，尤其對于需要長期用藥的慢性復(fù)雜性疾病更是如此。炎癥免疫反應(yīng)軟調(diào)節(jié)（SRIIR）是指藥物在控制IIR相關(guān)細胞異?；罨耐瑫r盡量減少對細胞生理功能的損害，即：行駛SRIIR的藥物不應(yīng)是對細胞功能、基因和蛋白表達或活性的完全抑制，而是在不同細胞上發(fā)現(xiàn)共性的或類似的信號轉(zhuǎn)導(dǎo)和網(wǎng)絡(luò)調(diào)控關(guān)鍵分子及特征，選擇性地將異常改變的細胞和分子活性調(diào)控至正常生理水平，恢復(fù)細胞功能的動態(tài)平衡，以發(fā)揮藥物的治療作用，且盡量減少不良反應(yīng)。SRIIR是研發(fā)治療IIR相關(guān)疾病創(chuàng)新藥物的新方向。

關(guān)鍵詞：炎癥免疫反應(yīng)；炎癥免疫反應(yīng)軟調(diào)節(jié)；信號轉(zhuǎn)導(dǎo)；新藥發(fā)現(xiàn)

基金項目：國家自然科學(xué)基金重點項目資助（81330081）

通訊作者：魏 偉，E-mail：wwei@ahmu.edu.cn，Tel：（0551）65161209

T3-6

二仙膏對化療后白細胞減少癥的藥效作用

許律捷1，康 德1，王金華1，劉艾林1，杜冠華1，杜新磊2

（1.中國醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院藥物研究所，北京100050；2.山東方健制藥有限公司）

摘要：目的研究探討傳統(tǒng)中藥二仙膏對環(huán)磷酰胺致小鼠白細胞減少癥的影響。方法①在治療實驗 昆明小鼠隨機分為對照組、模型組（環(huán)磷酰胺，100 mg·kg-1，ip）、二仙膏低、中、高三個劑量組（3，6，12 g·kg-1，ig）組、陽性藥組（瑞白，50 μg·kg-1，ih）。檢測小鼠的外周血白細胞數(shù)，股骨有核細胞數(shù)，計算脾臟指數(shù)和胸腺指數(shù)。在預(yù)防實驗中，昆明小鼠隨機分為對照組、模型組（環(huán)磷酰胺，200 mg·kg-1，ip）、二仙膏低、中、高三個劑量組（1.5，3，6 g·kg-1，ig）組、陽性藥組（瑞白，25 μg·kg-1，ih），對預(yù)防給藥5 d，造模后給藥7 d，檢測小-鼠外周血白細胞數(shù)，取材后檢測股骨有核細胞數(shù)，并計算脾臟指數(shù)和胸腺指數(shù)。結(jié)果在治療實驗中，與模型組相比較，二仙膏可以顯著提高脾指數(shù)，并顯著增加骨髓有核細胞數(shù)。②在預(yù)防實驗 二仙膏可以顯著增加外周血白細胞數(shù)和股骨有核細胞數(shù)，并顯著提高胸腺指數(shù)。結(jié)論二仙膏對于環(huán)磷酰胺致小鼠白細胞減少癥具有明顯的預(yù)防和治療作用。

關(guān)鍵詞：二仙膏；環(huán)磷酰胺；白細胞減少癥

基金項目：山東省科技重大專項項目（2015ZDXX0302A01）

通訊作者：劉艾林，E-mail：liuailin@imm.ac.cn，Tel：（010）63165184；杜冠華，E-mail：dugh@imm.ac.cn，Tel：（010）63165184；杜新磊，E-mail：Sdfjzy@163.com，Tel：（0537）2315804

T3-7

不同提取方法及給藥途徑的腸腑康膠囊對大鼠脾虛型潰瘍性結(jié)腸炎的影響

張小麗，陳瑞明，李 芳，楊智鋒，樊炳輝

（陜西省中醫(yī)藥研究院，陜西西安 710003）

摘要：目的探究不同提取方法及給藥途徑的腸腑康膠囊對大鼠脾虛型潰瘍性結(jié)腸炎（UC）的影響。方法選取正常大鼠84只，隨機分成6組：陰性對照組，模型組，口服給藥+直腸給藥組，分別提取口服給藥組，混合提取口服給藥組，補脾益腸丸陽性組。對照組正常飼養(yǎng)外，其余6組建立脾虛型潰瘍性結(jié)腸炎模型，將大鼠禁食不禁水12 h，灌服番瀉葉浸劑，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/只灌腸沖洗；動物蘇醒后，正常飼養(yǎng)，從第2天開始，繼續(xù)灌服番瀉葉浸劑2 d。在已復(fù)制的脾虛型模型基礎(chǔ)上，采用2，4，6-三硝基苯磺酸復(fù)制潰瘍性結(jié)腸炎模型：將大鼠禁食24 h后，用10%水合氯醛（350 mg·kg-1）腹腔注射麻醉，固定在鼠解剖臺上，準(zhǔn)備一直徑2.0 mm長約12 cm的橡膠輸液管，用液體石蠟潤滑后，輕緩插入肛門約8 cm，將50%乙醇與三硝基苯磺酸（TNBS，100 mg·kg-1）混合液0.25 mL注入大鼠結(jié)腸后注入0.3 mL空氣，再將大鼠提尾倒置5 min，讓其平躺自然清醒。于造模完成24 h后，在模型對照組中隨機選取2只大鼠進行解剖，肉眼觀察結(jié)腸黏膜組織損傷情況，并截取約1 cm左右病變結(jié)腸行病理組織學(xué)檢查，以確定模型復(fù)制是否成功。造模成功后，模型組和陰性對照組灌服水外，剩下4組按藥物劑量組別分別進行灌胃，15 d后，觀察大鼠癥狀，體征，生理病變。結(jié)果陰性對照組大鼠結(jié)腸光滑，無水腫及血絲，與陰性對照組比較，造模后大鼠出現(xiàn)嗜睡，懶動，蜷縮，飲少，納呆，精神萎靡，毛發(fā)疏松粗糙晦暗無光澤，腹瀉及黏液血便癥狀，大鼠體質(zhì)量明顯下降，攝食飲水量明顯降低，小便量明顯減少，大便濕重增加，自發(fā)活動次數(shù)明顯減少。造模后各組大鼠結(jié)腸黏膜組織在光鏡下可見到大量紅血絲、水腫、部分結(jié)腸潰瘍。給藥15 d后，解剖見給藥各組大鼠結(jié)腸水腫潰瘍數(shù)量明顯比模型組減少，其中口服+直腸給藥組結(jié)腸病變改善最為良好，水腫數(shù)量很少，潰瘍數(shù)及潰瘍面積最少，分別提取組次之，混合提取組不理想。病理組織學(xué)檢查：陰性組結(jié)腸大體正常，大腸腺體規(guī)則，可見大量胞質(zhì)富含黏液的杯狀細胞，黏膜固有層、黏膜下層未見炎性細胞浸潤。模型組腸黏膜可見潰瘍，可見“陷窩膿腫”、“陷窩潰瘍”形成，炎癥程度較重，可見腸黏膜和黏膜下組織大量炎細胞浸潤。陽性組結(jié)腸大體正常，大腸腺體規(guī)則，腸黏膜可見少量炎細胞浸潤?？诜?直腸組結(jié)腸大體正常，大腸腺體規(guī)則，腸黏膜可見少量炎細胞浸潤。分別提取腸黏膜可見炎細胞浸潤?；旌咸崛∧c黏膜可見輕度潰瘍，可見腸黏膜和黏膜下組織大量炎細胞浸潤。結(jié)論不同給藥途徑和提取方法的腸腑康膠囊對大鼠脾虛型潰瘍性結(jié)腸炎的影響不同，其中口服+直腸給藥療效最好，分別提取后給藥次之，混合提取后給藥效果不夠理想。

關(guān)鍵詞：提取方法；給藥途徑；腸腑康膠囊；脾虛型潰瘍性結(jié)腸炎

基金項目：陜西省科技統(tǒng)籌創(chuàng)新工程計劃項目（2014KTCQ03-10）

通訊作者：張小麗，E-mail：zhangxiaoli8788@163.com，Tel：13038932330

T3-8

白虎加桂枝湯對熱痹模型大鼠特征性甲基化基因表達的影響

陳 歡1，2，巨少華1，2，付文君1，2，魏江平1，2，徐世軍1，2

（1.四川省中藥資源系統(tǒng)研究與開發(fā)利用國家重點實驗室培育基地，四川成都 611137；2.成都中醫(yī)藥大學(xué)，四川成都611137）

摘要：目的考察白虎加桂枝湯對熱痹特征性甲基化基因的影響，揭示其治療熱痹的表觀遺傳學(xué)機制。方法采用佐劑性關(guān)節(jié)炎模型（AA）復(fù)合濕熱環(huán)境（溫度37℃、相對濕度70%~80%，2 h/d）制備熱痹模型（HB）。第15 d后，連續(xù)灌胃給與白虎加桂枝湯（28，14和7 g·kg-1）30 d后取空白組（NG）、AA及HB大鼠膝關(guān)節(jié)滑膜，采用MeDIP-Seq測序檢測滑膜DNA甲基化水平，以取NGvsHB和NGvsAA差集的方式篩選熱痹特征性甲基化基因，并采用qRT-PCR檢測特征性甲基化基因的表達水平。結(jié)果NGvsAA，NGvsHB及NGvsHB各有差異性甲基化基因分別為705，2418及1287個，這些甲基化差異性基因主要與炎癥免疫通路，如T、B細胞受體信號通路、MAPK信號通路及抗原提呈和處理等有關(guān)，生物合成與代謝通路，如多種氨基酸代謝途徑及糖類、蛋白質(zhì)代謝途徑等有關(guān)；熱痹特征性甲基化基因共42個，與AA比較，白虎加桂枝湯高、中、低各劑量組均能抑制熱痹特征性甲基化下調(diào)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）的表達，而僅高、中劑量組促進熱痹特征性甲基化上調(diào)基因Agxt（1.35±0.20，1.15±0.23）的表達水平。結(jié)論特征性基因甲基化水平變化可能是導(dǎo)致熱痹發(fā)病的原因，白虎加桂枝湯可通過糾正特征性基因的甲基化水平達到治療疾病的目的。

關(guān)鍵詞：類風(fēng)濕性關(guān)節(jié)炎；熱痹；DNA甲基化；白虎加桂枝湯

基金項目：國家自然科學(xué)基金面上項目（81273900）；中藥藥理四川省青年科技創(chuàng)新研究團隊（2014TD0007）

通訊作者：徐世軍，E-mail：docxu@126.com，Tel：（028）61800231

T3-9

雷公藤內(nèi)酯醇LLDT-8在抗GBM抗體誘導(dǎo)小鼠腎炎中的療效作用及對Fc受體介導(dǎo)淋巴細胞活化的調(diào)控機制

祁 青1，2，何世君2，唐 煒2，左建平1，2

（1.上海中醫(yī)藥大學(xué)免疫與病毒實驗室，上海 201203；2.中國科學(xué)院上海藥物研究所免疫藥理學(xué)研究室，上海201203）

摘要：目的研究（5R）-5-羥基雷公藤內(nèi)酯醇（LLDT-8）對小鼠抗腎小球基底膜（GBM）腎炎的療效作用，探討LLDT-8對Fcγ受體介導(dǎo)的淋巴細胞活化的調(diào)控機制。方法（1）運用兔抗小鼠GBM血清誘導(dǎo)的雄性NZW小鼠腎炎模型，以LLDT-8口服治療4周。觀察藥物對腎炎小鼠的體重、蛋白尿、腎病理變化等指標(biāo)的影響作用；（2）采用流式細胞術(shù)對LLDT-8治療干預(yù)腎炎小鼠腎浸潤細胞亞群比例及表型進行分析；（3）采用免疫熒光法觀察小鼠腎小球中補體沉積及炎性單核細胞浸潤；（4）采用PCR法檢測腎組織中Fcγ受體及炎性因子表達；（5）采用腎炎小鼠疾病高峰脾淋巴細胞，考察LLDT-8對抗血清誘導(dǎo)記憶性細胞應(yīng)答的影響。結(jié)果（1）LLDT-8能緩解抗GBM抗體誘導(dǎo)腎炎小鼠的疾病進程，改善腎病變，減少腎補體沉積及免疫細胞浸潤。（2）LLDT-8治療能降低疾病高峰血清中IFN-γ及IL-6水平。（3）LLDT-8降低疾病發(fā)展各階段的外周及腎CD11b+單核細胞Fcγ受體表達水平。（4）LLDT-8體外抑制腎炎小鼠淋巴細胞抗原特異性增殖，降低IgG及IL-6的分泌水平。結(jié)論證實了LLDT-8對于抗GBM抗體誘導(dǎo)的小鼠腎炎具有顯著的治療作用，闡明了LLDT-8影響單核/淋巴細胞的Fcγ受體的表達，調(diào)控抗原特異性記憶淋巴細胞應(yīng)答效應(yīng)并抑制腎病灶中免疫細胞浸潤，從而緩解腎小球腎炎進程的療效機制。

關(guān)鍵詞：雷公藤內(nèi)酯醇；抗GBM抗體誘導(dǎo)腎炎；Fcγ受體；淋巴細胞活化

基金項目：國家新藥創(chuàng)制重大專項（012014ZX09101002-001-02）

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

T3-10

DL1502新晶型對骨關(guān)節(jié)炎模型的治療作用

王曉波1，2，趙曉悅1，王海港1，于子茹1，楊玉林1，2，孫 悅2，王淑美2，杜冠華1，2

（1.中國醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院藥物研究所，北京市藥物靶點研究與新藥篩選重點實驗室，北京100050；2.廣東藥科大學(xué)，廣東廣州 510006）

摘要：目的觀察DL1502的新晶型對大鼠膝骨關(guān)節(jié)炎所導(dǎo)致的關(guān)節(jié)間隙狹窄的保護作用以及對腫脹、疼痛等癥狀的影響，并探討了DL1502藥物新的給藥方式的藥效作用。方法采用關(guān)節(jié)腔藥物注射法制備大鼠膝關(guān)節(jié)骨關(guān)節(jié)炎模型。對動物進行自主活動、電子壓痛、步態(tài)評測等行為學(xué)評分。測量大鼠膝關(guān)節(jié)直徑評判腫脹程度；應(yīng)用小動物活體成像儀系統(tǒng)拍攝X-Ray平片測量大鼠造模側(cè)膝關(guān)節(jié)的關(guān)節(jié)腔間隙變狹窄程度。結(jié)果與正常組相比，模型組大鼠膝關(guān)節(jié)電子壓痛閾值評分顯著降低（P＜0.05），而DL1502新晶型藥物1.125和4.5 mg·kg-1劑量組及新給藥方式組均可顯著改善疼痛閾值（P＜0.05，P＜0.05和P＜0.05）。DL1502新晶型1.125和4.5 mg·kg-1劑量組以及新給藥方式組可顯著減輕由骨關(guān)節(jié)炎所引起的腫脹（P＜0.01，P＜0.01和P＜0.05）。X-Ray平片拍攝結(jié)果顯示DL1502新晶型藥物1.125和4.5 mg·kg-1劑量組能顯著改善由骨關(guān)節(jié)炎導(dǎo)致的關(guān)節(jié)間隙狹窄程度（P＜0.01和P＜0.01），并呈劑量依賴性關(guān)系。新給藥方式組關(guān)節(jié)間隙狹窄情況也有改善（P＜0.05），并且狹窄值改善情況優(yōu)于DL1502新晶型1.125 mg·kg-1劑量組（P＜ 0.05），與4.5 mg·kg-1劑量組相比沒有顯著性差異。結(jié)論本研究采用經(jīng)典的藥物注射法制備大鼠膝關(guān)節(jié)骨關(guān)節(jié)炎模型，評價了DL1502新晶型對骨關(guān)節(jié)炎模型的治療作用研究。DL1502對藥物注射法所導(dǎo)致的大鼠膝關(guān)節(jié)關(guān)節(jié)間隙變窄有著良好的保護作用，并對骨關(guān)節(jié)炎所致的腫脹、疼痛癥狀有改善作用。新的給藥方式同樣有著顯著的藥效作用。

關(guān)鍵詞：骨關(guān)節(jié)炎；新晶型；新給藥方式

通訊作者：杜冠華，E-mail：dugh@imm.ac.cn，Tel：（010）63165184

T3-11

神經(jīng)酰胺類鞘脂及其代謝通路參與免疫性肝纖維化發(fā)生發(fā)展的作用研究

李芳芳，劉 寧，劉 威，鮑秀琦，張 丹，孫 華

（中國醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院藥物研究所，北京100050）

摘要：目的探討神經(jīng)酰胺類鞘脂及其代謝通路在慢性免疫性肝纖維化發(fā)生發(fā)展中的作用。方法SD大鼠隨機分為兩組，模型組大鼠腹腔注射豬血清（PS）0.5 mL/次，每周2次，連續(xù)注射10周，誘導(dǎo)大鼠慢性免疫性肝纖維化，空白對照組大鼠注射等劑量生理鹽水。利用血清生化指標(biāo)、肝臟病理檢測、超聲影像檢查及肝組織羥脯氨酸含量等判斷肝纖維化形成及程度。HPLC-MS/MS分析血清鞘脂、特別是神經(jīng)酰胺類鞘脂的含量；Western蛋白印跡法檢測鞘脂代謝通路主要代謝酶的蛋白表達情況。結(jié)果SD大鼠給予PS后第4周，肝臟出現(xiàn)輕度纖維化。在第6，8周，肝超聲檢查出現(xiàn)纖維化造影，血清透明質(zhì)酸和肝羥脯氨酸的含量較空白對照組顯著升高。至第10周，可見假小葉形成。在肝纖維化發(fā)生發(fā)展過程中，血清多個鞘脂類成分，包括神經(jīng)酰胺、糖基化神經(jīng)酰胺含量發(fā)生明顯改變。在神經(jīng)酰胺的變化中，Cer（18∶1）含量在第4周顯著降低，后升高但與空白對照組比較仍顯著降低；Cer（22∶0），Cer（24∶1）的含量在第6周明顯升高，至第8周、第10周，含量顯著升高。Western蛋白印跡法檢測神經(jīng)酰胺代謝通路主要代謝酶的表達變化，神經(jīng)酰胺合成酶2（CerS2）表達顯著升高。結(jié)論神經(jīng)酰胺類鞘脂可能參與了慢性免疫性肝纖維化的發(fā)生發(fā)展，CerS2可能發(fā)揮重要作用，進一步的機制研究正在進行中。

關(guān)鍵詞：肝纖維化；神經(jīng)酰胺；神經(jīng)酰胺合成酶2

基金項目：國家科技計劃863項目（2014AA021101）；國家自然科學(xué)基金項目（81573487）

通訊作者：孫 華，E-mail：sunhua@imm.ac.cn，Tel：（010）63165203

T3-12

構(gòu)樹果汁體外抗氧化及對小鼠免疫調(diào)節(jié)作用

龐素秋2，金孝勤2，王國權(quán)1

（1.華僑大學(xué)生物醫(yī)學(xué)學(xué)院，福建泉州 362021；2.中國人民解放軍第180醫(yī)院，福建泉州 362000）

摘要：目的探討構(gòu)樹果汁（BPFJ）體外抗氧化作用及對小鼠的免疫活性影響。方法從清除超氧陰離子自由基（O2-·）、羥基自由基（·OH）、DPPH自由基等方法觀察了BPFJ體外抗氧化作用；用刀豆蛋白（ConA）誘導(dǎo)小鼠脾淋巴T細胞增殖細胞模型，MTT法測定細胞增殖并檢測BPFJ對小鼠脾T淋巴細胞增殖的影響。結(jié)果BPFJ能顯著清除O2-·、·OH和DPPH自由基，在0.1～2.0 g·L-1范圍內(nèi)呈現(xiàn)劑量效應(yīng)關(guān)系。小鼠T淋巴細胞在ConA刺激下48 h后，MTT吸收度顯著高于正常對照組。而BPFJ組與ConA組比較則顯著降低了MTT吸收度。結(jié)論BPFJ有較強的體外抗氧化作用并能顯著提高小鼠的免疫作用。BPFJ是一種抗氧化功效較強的活性物質(zhì)，具有很好的保健功能。

關(guān)鍵詞：構(gòu)樹果汁；抗氧化；自由基；免疫調(diào)節(jié)

基金項目：福建省泉州市科技項目（200712Z）

通訊作者：王國權(quán)，E-mail：wangguoqun@hqu.edu.cn

T3-13

清除衰老細胞治療小鼠放射性肺纖維化機制

潘 金1，徐燕鋒1，張俊伶2，李德冠2，王月英2，陳孟毅1，劉 冰1，周道洪3，孟愛民1

（1.中國醫(yī)學(xué)科學(xué)院協(xié)和醫(yī)學(xué)院，醫(yī)學(xué)實驗動物研究所，北京協(xié)和醫(yī)學(xué)院比較醫(yī)學(xué)中心，北京 100221；2.中國醫(yī)學(xué)科學(xué)院放射醫(yī)學(xué)研究所，天津市放射醫(yī)學(xué)分子核醫(yī)學(xué)重點實驗室，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：目的放射性肺纖維化是放射性肺損傷的晚期改變，發(fā)病機制復(fù)雜，臨床上缺乏有效的治療方法，患者預(yù)后差。對放射性肺纖維化機制及干預(yù)的研究是當(dāng)前的研究熱點。ABT-263是抗腫瘤新藥，近期發(fā)現(xiàn)具有清除衰老細胞、改善造血干細胞輻射損傷的作用。研究在前期研究的基礎(chǔ)上建立局部照射誘導(dǎo)的肺纖維化小鼠模型，觀察ABT-263對放射性肺纖維化的治療作用，探討其作用機制。方法健康雄性C57BL/6J小鼠分為對照組（Control組）、給藥組（ABT組）、照射組（IR組）和治療組（IR+ABT組）。小鼠接受17Gy右側(cè)胸部X射線照射，照射后第16周開始灌胃給予ABT-263 50 mg·kg-1，每療程給藥5 d，停藥2周，給藥2個療程后進行觀察。兩個療程治療后CT檢查小鼠肺實質(zhì)的損傷情況。病理觀察小鼠肺組織結(jié)構(gòu)的變化、膠原生成（Masson檢測）和β-半乳糖苷酶（β-Gal）表達。免疫熒光檢測肺組織中肺表面活性前體蛋白C（Pro-SPC）和P16的表達?；騊CR芯片技術(shù)檢測肺組織纖維化相關(guān)基因的表達。結(jié)果胸部CT結(jié)果顯示ABT治療可以降低放射引起的小鼠肺組織的HU值升高（P＜0.05）；降低照射引起小鼠肺臟指數(shù)顯著升高（P＜0.05）；減輕受照小鼠肺組織病理變化評分（P＜0.01），減少膠原沉積面積（P＜0.01），降低β-Gal表達細胞比例（P＜0.01）；可以抑制受照小鼠Ⅱ型肺泡細胞P16表達（P＜0.01）；纖維化相關(guān)基因芯片測定結(jié)果顯示ABT可以降低受照小鼠肺組織中Bcl-2、Col3a1、Col1a2、Ⅱ1α、Il1β mRNA顯著升高（P＜0.05或P＜0.01）。結(jié)論上述研究結(jié)果顯示ABT263可以減輕小鼠放射性肺纖維化的損傷性改變，降低衰老Ⅱ型肺上皮細胞，抑制Bcl-2及肺組織炎癥性因子及纖維化相關(guān)因子的表達。研究結(jié)果將為放射性肺纖維化防治提供新的思路。

關(guān)鍵詞：放射性肺損傷；肺纖維化；細胞衰老；組織干細胞；Ⅱ型肺泡上皮細胞

基金項目：國家自然科學(xué)基金（81573094；81372928）

通訊作者：孟愛民，E-mail：ai_min_meng@126.com，Tel：13911809579

T3-14

異甘草酸鎂對小鼠放射性肺纖維化的改善作用及其機制的初步探討

張 攀，劉 濤，王芳芹，陽群芳，陳曉紅

（第三軍醫(yī)大學(xué)藥學(xué)系藥理教研室，重慶 400038）

摘要：目的探討異甘草酸鎂（MgIG）對放射性肺纖維化的作用及其抗纖維化的機制。方法將SPF級雌性C57小鼠隨機分為正常對照組、輻照模型組、異甘草酸鎂對照組、異甘草酸鎂治療組和地塞米松治療組，每組10只。除正常對照組和異甘草酸鎂對照組，其余各組小鼠均給予單次15 Gy60Coγ射線全肺照射，照射后12周處死小鼠取左肺下葉組織，免疫組化法觀察肺組織Ⅰ型、Ⅲ型膠原蛋白及轉(zhuǎn)化生長因子-β1（TGF-β1）信號通路蛋白表達變化。結(jié)果各組小鼠肺組織均有Ⅰ型、Ⅲ型膠原及TGF-β1，Psmad2，Psmad3不同程度的表達，模型組明顯高于正常對照組，異甘草酸鎂治療組與地塞米松治療組相較于模型組各蛋白表達均有下降。結(jié)論異甘草酸鎂可通過抑制小鼠肺組織TGF-β1信號通路改善放射性肺纖維。

關(guān)鍵詞：放射性肺纖維化；異甘草酸鎂；轉(zhuǎn)化生長因子β1

基金項目：國家自然科學(xué)基金項目（81272995）

通訊作者：陳曉紅，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，F(xiàn)oxp3 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，F(xiàn)ANG 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，F(xiàn)uwai 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

吉林省科技發(fā)展計劃項目（20140204060YY）

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

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