抗血管紧张素ⅡAT1受体抗体对大鼠脾脏T淋巴细胞的促增殖作用  

Antibody against angiotensin AT1 receptor promotes proliferation of T lymphocytes from rats＇ spleen

作　　者：张苏丽[1] 杨丽红[2] 郑荣华[1] 李晓宇[1] 闫莉[1] 武烨[1] 贺忠梅[1] 刘慧荣[3] 

机构地区：[1]山西医科大学生理教研室山西医科大学细胞生理学省部共建教育部重点实验室,太原市030001 [2]山西医科大学病理教研室 [3]首都医科大学心脏病学系

出　　处：《中国心血管病研究》2009年第2期135-138,共4页Chinese Journal of Cardiovascular Research

基　　金：太原市2008年大学生创新创业专项项目（08122033）;北京市属市管高等学校人才强教计划资助项目（10850302）;山西省基础研究项目（2008011076-2）

摘　　要：目的观察抗AT1受体（AT1R）抗体对培养的大鼠脾脏T淋巴细胞增殖活性的影响。方法以人工合成的AT1R细胞外第二环肽段（AT1R—ECⅡ）为抗原主动免疫Wistar大鼠，利用亲和层析法提纯免疫大鼠血清中含有抗AT1R抗体的IgGs；培养大鼠脾淋巴细胞，刀豆蛋白A（ConA）诱导其活化和增殖，分别给予不同浓度的提纯IgGs（0．01、0．1、1umol／L）和血管紧张素Ⅱ进行干预，通过CCK-8法测定脾淋巴细胞增殖情况，并确定抗AT，R抗体的作用位点。结果ConA刺激48h后，大鼠脾淋巴细胞CCK-8吸光度显著高于空白对照组（0．38±0．05比0．27±0．02，P〈0．05）；不同浓度的IgGs剂量依赖性促进脾脏T淋巴细胞增殖（A值分别为0．53±0．03、0．71±0．04和0．94±0．05），与血管紧张素Ⅱ的作用相类似（A值为0．73±0．14、1．52±0．17和2．14±0．12）；AT1R特异性阻断剂Losartan和AT，R—ECⅡ均可显著减弱含抗AT1R抗体IgGs的促增殖效应（A值由0．71±0．04分别降为0．54±0．02，P〈0．01；0．52±0．04，P〈0．01）。结论抗AT1R抗体具有受体激动剂样效应，通过特异结合AT1R—ECⅡ剂量依赖性增强离体脾T淋巴细胞增殖活性。Objective To investigate the effect of antibody against angiotensin Ⅱ AT1 receptor （AT1-RA） on proliferation of T lymphoeytes from rats＇ spleen in vitro. Methods Health adult rats were actively immunized with the synthetic peptide corresponding to the sequences for the second extracellular loop of AT1 receptor （AT1R- EC Ⅱ ）. The total IgGs in immunized rats＂ serum, containing quantity of AT1-RA, were purified with anti-rat IgG affinity column. Lymphoeytes isolated from rats＇ spleen and stimulated by Con A were treated with different concentrations of AT1-RA （0.01 umol/L,0.1 umol/L, 1 umol/L） or angiotensin II respectively, CCK-8 was used to detect the lymphocytes＇ proliferation response. In addition, the binding site of AT1-RA reacting with lymphocytes was determined. Results After 48 h, CCK-8 A value of lymphoeytes that reacted with Con A increased significantly compared with that of cells without Con A. All three different concentrations of AT1-RA could enhance the proliferation response of T lymphocytes from rats＂ spleen （A value,0.53±0.03,0.71±0.04, 0.94±0.05）. These effects of AT1-RA were almost identical with that of angiotensin Ⅱ （A values,0.73±0.14, 1.52±0.17,2.14±0.12）. Either Losartan or AT1R-EC Ⅱ attenuated the proliferation response induced by AT1-RA （A value decreased from 0.71± 0.06 to 0.54±0.02,P〈0.01 ;0.52±0.04, P〈0.01 ,respectively）. Conclusion Antibody against AT1R presenting an agonist-like activity could promote proliferation of T lymphocytes from rats＇ spleen in a dose-dependent fashion by interacting with AT1R-EC Ⅱ specially.

关 键 词：血管紧张素Ⅱ1型受体 抗体 T淋巴细胞 细胞增殖 

分 类 号：Q95-33[生物学—动物学] R54[医药卫生—心血管疾病]