體育學系暨研究所2018-04-172025-07-272018-04-172018-04-17https://ir.ntus.edu.tw/handle/987654321/63609計畫編號：NSC97-2410-H028-002 研究期間：2008/08/01~2009/07/31為了了解競技自行車選手經中強度運動訓練之後，是否具有較強之免疫刺激 反應以抗B 型肝炎病毒之表現； 分組抽取人類周邊血液單核球細胞以 phytohemagglutinin剌激後；收集已去除單核球細胞之調節培養基後加入能表現 B型肝炎病毒表面抗原之Hep3B細胞株。此為新的作為研究運動，免疫系統和病毒 之間作用之ex vivo模式系統。在我們預備試驗的數據中顯示經中強度運動訓練 之後競技自行車選手其調節培養基對Hep3B細胞株之病毒表面抗原的表現有較大 之抑制作用；其在5μg/ml 之phytohemagglutinin刺激下可達61.5%之表現抑制作 用而比較沈靜控制組則為85.8%。其調節培養基有較多之IL-1β之分泌；其在 5μg/ml 之phytohemagglutinin刺激下只達352 ± 43 pg/ml之表現抑制作用而比 較沈靜控制組則為830 ± 59 pg/ml。 而此種病毒表面抗原的表現抑制作用應為免疫細胞在調節培養基中經刺激 所產生之各式細胞激素所造成。故檢測各式細胞激素(如IFN-γ, TNF-α, IFN-α and IL-1β)和PHA劑量關係在兩組間之差異以釐清中強度運動訓練之具有較強 之免疫調節能力以抗病毒表面抗原的表現。以抗各式細胞激素抗体加入之中和試 驗以釐清那種細胞激素為此抑制作用之主要作用耇。單離出各種免疫細胞(如 CD3+, CD4+, CD8+, CD14+, pDC (BDCA4+), mDC (BDCA1+) 或 CD56+)後再經 排列組合並且觀察其抑制作用及細胞激素分泌情況以了解何種免疫細胞所刺激 產生之主要作用細胞激素。 並且以細胞內細胞激素檢測法進一步地証實。再進一步以reverse transcription (RT), polymerase chain reaction (PCR) and proteomic analysis等法釐清之前所証實之作用免疫細胞內因運動所誘發之胞內分子調控 機制。 To evaluate whether competitive cyclists with moderate training intensity have greater immune response against hepatitis B virus than sedentary controls, human peripheral blood mononuclear cells (MNC) will be stimulated by phytohemagglutinin to prepare conditioned media (MNC-CM) for assessment of the inhibitory effects on hepatitis B surface antigen (HBsAg) expression in human hepatoma Hep3B cells. It is a novel ex vivo model system to elucidate the relationship between the exercise immunology and anti-virs activity. Our preliminary experimental results showed that the inhibitory effect against the relative HBsAg expression of cyclists’ MNC-CM was greater than that of control. For example, the cyclists’ MNC-CM prepared from 5 μg/ml phytohemagglutinin decreased HBsAg expression to 61.5% whereas that of controls remained at 85.8%. The cyclists’ MNC-CM prepared from 5 μg/ml phytohemagglutinin to secret the amounts of IL-1β, in CMT-PHA-MNC-CM increased up to 830 ± 59 pg/ml compared to those in SC-PHA-MNC-CM at 352 ± 43 pg/ml. The relationship between phytohemagglutinin dose-dependent and levels of cytokines such as IFN-γ, TNF-α, IFN-α and IL-1β in the MNC-CM from the cyclists and control group will be compared to elucidate the cytokines secretion of immunomodulation by moderate exercise. Antibody neutralization of cyclists’ MNC-CM and addition of recombinant cytokines into controls’ MNC-CM will be measured to elucidate whether IFN-γ, TNF-α and IFN-α had synergistic effects against HBsAg expression. To clarify the cytokines secreting related cells and to clarify the interaction between the lymphocytes, isolation of possible cytokines secreting lymphocytes and combination of each identical lymphocyte (such as CD3+, CD4+, CD8+, CD14+, pDC (BDCA4+), mDC (BDCA1+) or CD56+) will be performed. Furthermore, .intracellular cytokines assay will be carried out to clarify the cytokines secreting related cells and to clarify the interaction between the lymphocytes. To evaluate the molecular mechanism of promoting cytokines secreting in moderate exercise, reverse transcription (RT), polymerase chain reaction (PCR) and proteomic analysis will be performed to evaluate the gene expression about cytokines secretion promoting by moderate exercise.383688 bytesapplication/pdf中度強度運動; 自行車選手; 免疫調節; B型肝炎病毒; B型肝炎病毒表面抗原cyclists; immunomodulatory; HBV; HBsAgreport