摘要
本文探讨了精氨酸血管升压素(AVP)刺激下体外培养的大鼠心肌成纤维细胞(CFs)内一氧化氮(NO)含量、一氧化氮合酶(NOS)活性、诱导型一氧化氮合酶基因表达的变化及其与核因子κB(NF-κB)的关系。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、逆转录一聚合酶链式反应(RTPCR)、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NFKB的活化。结果显示,AVP浓度依赖性(0.001—0.1μmol/L)地增加CFs的NO含量,提高NOS活性,增加iNOSmRNA表达;AVP能够活化NF-κB,使其由细胞浆转位于细胞核;NF-κB特异性抑制剂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CFsiNOS mRNA表达增加、NOS活性增高、NO合成增多可能通过NF-κB激活途径,NF-κB激活参与心肌纤维化的发生和发展。
To investigate the changes in the nitric oxide(NO) contents, nitric oxide synthase(NOS) activity and inducible nitric oxide (iNOS) mRNA expression in arginine vasopressin (AVP)-induced cardiac fibroblasts (CFs) in vitro and its relation to nuclear factor-κB(NF-κB), CFs were isolated by trypsin digestion method. 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-κB in CFs. AVP increased NO contents, NOS activity and iNOS mRNA expressions in a concentration-dependent manner; NF-κB was activated and mobilized from cytoplasm to nucleus in AVP-induced CFs; PDTC,one of the inhibitors of NF-κB, 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-κB activation pathway in cultured CFs induced by AVP, and that NF-κB is involved in the occurrence and development of myocardial fibrosis.
出处
《生理学报》
CAS
CSCD
北大核心
2003年第4期417-421,共5页
Acta Physiologica Sinica
基金
This work was supported by the National Natural Science Foundation of China (No. 39670317).
