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    流式細胞術檢測脂多糖對羅氏沼蝦血細胞活性和免疫功能的影響
    Flow cytometric analysis of viability and immune response of Macrobrachium rosenbergii haemocytes activated by lipopolysaccharide
    投稿時間:2009-04-19  修訂日期:2010-08-06
    DOI:
    中文關鍵詞:  脂多糖  羅氏沼蝦  血細胞  免疫反應  流式細胞術
    英文關鍵詞:Lipopolysaccharide  Macrobrachium rosenbergii  Haemocyte  Immune response  Flow cytometry
    基金項目:國家自然科學基金項目(面上項目,重點項目,重大項目)
    作者單位郵編
    冼健安 華南師范大學生命科學學院 510631
    王安利 華南師范大學生命科學學院 510631
    苗玉濤 華南師范大學生命科學學院 
    伍娟 華南師范大學生命科學學院 
    摘要點擊次數: 3769
    全文下載次數: 511
    中文摘要:
          以脂多糖(LPS)刺激羅氏沼蝦(Macrobrachium rosenbergii)離體血細胞,應用流式細胞術檢測血細胞大小、活性、吞噬功能和呼吸爆發的變化,并檢測細胞培養液中的酚氧化酶活力。根據前向角散射光(FSC)和側向角散射光(SSC)特征的不同,可把羅氏沼蝦的血細胞分為三個亞群:透明細胞、小顆粒細胞和大顆粒細胞。血細胞經LPS 刺激20-40min后,大型血細胞(小顆粒細胞和大顆粒細胞)的比例開始迅速下降,而細胞的大量死亡從40-60min開始。添加胰蛋白酶抑制劑(TI)能顯著減緩LPS導致的細胞變小和死亡。3和5µg/ml LPS能顯著提高血細胞的吞噬活力、呼吸爆發水平和培養液中的酚氧化酶活力。結果表明,LPS是羅氏沼蝦血細胞的強免疫刺激物和毒物,它刺激血細胞脫顆粒釋放酚氧化酶,提高了血細胞的吞噬能力,促進了大量活性氧的產生;其毒性致血細胞死亡可能是血細胞被誘導產生的過量活性氧帶來的氧化傷害所造成的。
    英文摘要:
          Cell size, viability, phagocytosis activity and respiratory burst (RB) of lipopolysaccharide (LPS) stimulated haemocytes from Macrobrachium rosenbergii were measured by flow cytometry. Phenoloxidase (PO) activity of the cell supernatant was also determined. According to the forward light scatter (FSC) and the side light scatter (SSC) signals of each individual, haemocytes could be divided into three subpopulations: hyaline cells, semigranular cells and granular cells. After 20-40 min of incubation with LPS, the percentage of large cells (semigranular and granular cells) decreased sharply, while the number of live cells began to reduce after 40-60 min. Trypsin inhibitor (TI) further delayed these changes induced by LPS. Phagocytosis activity, RB of haemocytes and PO activity of haemocyte supernatants significantly increased in the 3 and 5µg/ml LPS treatments. These results indicated that LPS was a stronge stimulator and toxicant for the M. rosenbergii haemocytes. The immune activity of haemocytes was activated by LPS. Semigranular and granular cells were activated to degranulation to release PO. LPS was also an efficient activator of phagocytosis and RB. It’s toxicity on the haemocytes may resulted from the damage of excessive ROS.
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