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| 树舌荧光多糖的制备及在小鼠脾淋巴细胞中的定位 |
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苏玲1, 胡静1, 李雨婷1, 金周雨1, 杨扬1, 杨颜铭1, 宋慧1,2
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| (1.吉林农业大学 生命科学学院, 长春 130118;2.食药用菌教育部工程研究中心, 长春 130118) |
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| 摘要: |
| 为确定树舌多糖在小鼠脾淋巴细胞上的定位,应用胺化还原法在树舌多糖(GAP)的还原性末端连接异硫氰酸荧光素(FITC),用荧光分光光度计测定其荧光取代度,xCELLigence RTCA DP全自动实时细胞分析仪检测GAP荧光标记前后生物学稳定性。流式细胞仪检测小鼠脾T淋巴细胞、B淋巴细胞和NK细胞中的GAP荧光强度,荧光显微镜观察GAP在小鼠脾淋巴细胞中的定位。结果显示:在荧光标记GAP(FITC-GAP)前后的细胞毒活性不变,并且其荧光取代率为0.90%;流式细胞仪检测发现小鼠脾T淋巴细胞、B淋巴细胞和NK细胞上的荧光信号强度与FITC-GAP加入量成正相关,荧光显微照片显示GAP定位于小鼠脾淋巴细胞表面并可转运至细胞核。 |
| 关键词: 树舌 多糖 荧光标记 脾细胞 细胞定位 |
| DOI：10.11841/j.issn.1007-4333.2013.01.022 |
| 投稿时间：2012-04-25 |
| 基金项目:吉林省世行贷款农产品质量安全项目(2011-Y18) |
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| Preparation of fluorescent polysaccharides from Ganoderma applanatum and cellular localization on splenic lymphocytes of mice |
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SU Ling1, HU Jing1, LI Yu-ting1, JIN Zhou-yu1, YANG Yang1, YANG Yan-ming1, SONG Hui1,2
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| (1.School of Llife Science, Jilin Agriculture University, Changchun 130118, China;2.Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi, Jilin Agricultural University, Changchun 130118, China) |
| Abstract: |
| In order to confirm cellular localization on splenic lymphocytes of the mice,fluorophores was used to polysaccharide from Ganoderma applanatum (GAP)and fluorescence signal was detected indirectly to trace GAP.The reductive amination method was used in fluorescently tagged of Ganoderma applanatum polysaccharide(GAP).Fluorescein isothiocyanate was linked with reducibility terminatio of GAP,and fluorescent substitute ratio is detected by fluorescence spectrophotometer.The biological stability of GAP before and after fluorescently-tagged was measured by xCELLigence RTCA DP automatic real-time cell analyzer.The fluorescence signal of T lymphocytes,B lymphocytes and natural killer cell of spleen were detected by flow cytometry,and cellular localization was observed by fluorescence microscope.The results showed that there is no significant changes for the cytotoxic activity in vitro before and after fluorescent labeling of GAP,and its fluorescent substitute ratio is 0.90%.The results flowing cytometry showed that fluorescence signal could be detected by each FITC-GAP experimental group of T lymphocytes,B lymphocytes and natural killer cell of mice,and fluorescence signal intensity increased with the increased FITC-GAP concentration.Observed by fluorescence microscopy,FITC-GAP both surface and intracellular.GAP was located on the cell surface and transported to the cell nucleus of splenic lymphocytes for mice. |
| Key words: Ganoderma applanatum polysaccharide fluorescent labeling spleen lymphocytes cellular localization |
