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核因子#kappa#B在精氨酸血管升压素诱导大鼠心肌成纤维细胞一氧化氮合成中的作用

范延红, 赵连友, 郑强荪, 薛玉生, 杨学东, 田建伟, 徐琳

第四军医大学西京医院心脏内科.陕西,西安 710033；第四军医大学唐都医院心脏内科.陕西,西安 710038

摘要

用胰酶消化法分离培养Sprague Dawley仔鼠的CFs,分别采用硝酸还原酶法、分光光度法、逆转录-聚合酶链式反应(RT-PCR)、免疫荧光-共聚焦显微镜和蛋白质印迹检测AVP干预下CFs的NO含量、NOS活性、iNOS mRNA表达和NF-#kappa#B的活化。结果显示,AVP浓度依赖性(0.001--0.1#mu#mol/L)地增加CFs的NO含量,提高NOS活性,增加iNOS mRNA表达;AVP能够活化NF-#kappa#B,使其由细胞浆转位于细胞核;NF-#kappa#B特异性抑制吡咯啉烷二甲基硫脲(PDTC)能够抑制AVP诱导的CFs NO含量增加、NOS活性提高和iNOS mRNA表达增加。上述结果提示,AVP干预下CFs iNOS mRNA表达增加、NOS活性增高、NO合成增多可能通过NF-#kappa#B激活途径,NF-#kappa#B激活参与心肌纤维化的发生和发展。

关键词： 精氨酸升压素; 一氧化氮; 心肌成纤维细胞; 核因子κB

Arginine vasopressin-induced nitric oxide content changes in cultured cardiac fibroblasts and its relation to nuclear factor-#kappa#B

Fan Yanhong, Zhao Lianyou, Zheng Qiangsun, Xue Yusheng, Yang Xuedong, Tian Jianwei, Xu Lin

Department of Cardiology,Xijing Hospital,Fourth Military Medical University.Xi^an 710033,Shaanxi；China

Abstract

Nitric acid reductase method, spectrophotometry, reverse transcription-polymerase chain reaction (RT-PCR), immunofluorescence-interactive laser cytometer techniques and Western blotting were used respectively to detect NO contents, NOS activity, iNOS mRNA expression and the activation of NF-#kappa#B in CFs. AVP increased NO contents, NOS activity and iNOS mRNA expressions in a concentration-dependent manner; NF-#kappa#B was activated and mobilized from cytoplasm to nucleus in AVP-induced CFs; PDTC, one of the inhibitors of NF-#kappa#B, could inhibit aforementioned increments. It is suggested that the increases in NO contents, elevation of NOS activity and increment of iNOS mRNA expression may be mediated through NF-#kappa#B activation pathway in cultured CFs induced by AVP, and that NF-#kappa#B is involved in the occurrence and development of myocardial fibrosis.

Key words: Arginine vasopressin;Nitric oxide;Cardiac fibroblasts;NF-κB

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引用本文：

范延红, 赵连友, 郑强荪, 薛玉生, 杨学东, 田建伟, 徐琳. 核因子#kappa#B在精氨酸血管升压素诱导大鼠心肌成纤维细胞一氧化氮合成中的作用[J]. 生理学报 2003; 55 (4): .

Fan Yanhong, Zhao Lianyou, Zheng Qiangsun, Xue Yusheng, Yang Xuedong, Tian Jianwei, Xu Lin. Arginine vasopressin-induced nitric oxide content changes in cultured cardiac fibroblasts and its relation to nuclear factor-#kappa#B. Acta Physiol Sin 2003; 55 (4): (in Chinese with English abstract).