枣多糖对小鼠脾脏B淋巴细胞体外增殖的影响

doi:10.3969/j.issn.1006-267x.2020.09.049

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摘要本试验旨在探究不同乙醇浓度提取的枣多糖（JP）协同脂多糖（LPS）对小鼠脾脏B淋巴细胞体外增殖的影响。本试验采用水提醇沉法和分步醇沉法获取4种JP；采用机械法无菌分离出小鼠脾脏B淋巴细胞，调节细胞浓度为5×10⁴个/mL，将细胞随机分为18组，每组4个重复，每个重复85 μL细胞。试验采用4×4双因子试验设计，乙醇浓度和JP浓度为2个主效应，乙醇浓度分别为60%、70%、80%和90%，JP浓度分别为25、50、100和150 μg/mL，另外设置1个空白对照组（不添加JP和LPS），1个LPS对照组（不添加JP，只添加LPS），用四甲基偶氮唑盐比色（MTT）法检测各组脾脏B淋巴细胞的增殖情况，并计算其相对增殖率。结果表明：1）B淋巴细胞的相对增殖率随着乙醇浓度的增加而增加，乙醇浓度为90%组的相对增殖率显著高于60%、70%和80%组（P<0.05），70%和80%组的相对增殖率显著高于60%组（P<0.05），且70%和80%组间没有显著差异（P>0.05）。2）随着JP浓度的增加，相对增殖率先增加后降低，各试验组均显著高于LPS对照组和空白对照组（P<0.05），且各试验组间差异显著（P<0.05），当JP浓度为100 μg/mL时相对增殖率最大。3）乙醇浓度和JP浓度两者互作对小鼠脾脏B淋巴细胞相对增殖率具有显著影响（P<0.05），LPS对照组和各试验组的相对增殖率均显著高于空白对照组（P<0.05），除乙醇浓度60%、JP浓度25 μg/mL组的相对增殖率显著低于LPS对照组（P<0.05），其他各组均显著高于LPS对照组（P<0.05）。在体外培养条件下，小鼠脾脏B淋巴细胞的相对增殖率随乙醇浓度的增加而增加，随JP浓度的增加呈"钟罩型"变化，饱和浓度为100 μg/mL；JP浓度和乙醇浓度互作可显著促进小鼠脾脏B淋巴细胞增殖，且乙醇浓度为80%、JP浓度为100 μg/mL时效果最好。

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	郭林霞
	李树鹏
	马可为
	郝艳霜
	赵国先
	杨博文
	郑紫薇
	张达
	张思轩
	冯志华
	刘观忠

关键词 ：枣多糖, 脾脏B淋巴细胞, 脂多糖, 相对增殖率

Abstract：The purpose of this experiment was to explore the effects of jujube polysaccharide (JP) extracted with different ethanol concantrations and cooperated with lipopolysaccharide (LPS) on the proliferation of mouse spleen B lymphocytes in vitro. In this experiment, four jujube polysaccharides were obtained by water extraction and alcohol precipitation method and step-by-step alcohol precipitation method in turn. Mouse spleen B lymphocytes were sterilely separated by mechanical method. The cells were adjusted to 5×10⁴ cells/mL and randomly divided into 18 groups with 4 replicates in each group and 85 μL cells in each replicate. The experiment adopted a 4×4 two-factor experimental design. The concentrations of ethanol and JP were two main effects. The concentration of ethanol were 60%, 70%, 80% and 90%, respectively, and the concentrations of JP were 25, 50, 100 and 150 μg/mL, respectively. In addition, one group of blank control (without addition of JP and LPS) and one group of LPS control (without addition of JP and only addition of LPS) were set up. The proliferation of B lymphocytes in each group was detected by MTT method, and the relative proliferation rate was calculated. Results showed as follows:1) the relative proliferation rate of spleen B lymphocytes increased with the increase of ethanol concentration, the relative proliferation rate of the 90% ethanol concentration group was significantly higher than that of the 60%, 70% and 80% ethanol concentration groups (P<0.05), the relative proliferation rate of the 70% and 80% ethanol concentration groups was significantly higher than that of the 60% ethanol concentration group (P<0.05), and there was no significant difference between the 70% and 80% ethanol concentration groups (P>0.05). 2) With the increase of JP concentration, the relative proliferation rate first increased and then decreased. All the test groups were significantly higher than LPS control group and blank control group (P<0.05), and the differences among the test groups were significant (P<0.05). When JP concentration was 100 μg/mL, the relative proliferation rate was the largest. 3) The interaction of ethanol concentration and JP concentration had a significant effect on the relative proliferation rate of mouse spleen B lymphocytes (P<0.05). The relative proliferation rate of LPS control group and each test groups was significantly higher than that of blank control group (P<0.05). The relative proliferation rate of 60% ethanol concentration and 25 μg/mL JP concentration group was significantly lower than that of LPS control group (P<0.05), while that of other groups was significantly higher than that of LPS control group (P<0.05). Under the condition of in vitro culture, the relative proliferation rate of mouse spleen B lymphocytes increases with the increase of ethanol concentration, and shows a "bell-jar" change with the increase of JP concentration, with a saturation concentration of 100 μg/mL. The interaction of JP concentration and ethanol concentration can significantly promote the proliferation of spleen B lymphocytes in mice, and the best effect is obtained when the ethanol concentration is 80% and JP concentration is 100 μg/mL.

Key words： jujube polysaccharide splenic B lymphocyte lipopolysaccharide relative proliferation rate

收稿日期: 2020-03-14

PACS:

S811.3

基金资助:河北省自然科学基金（C2019204330）；河北省现代农业产业技术体系蛋肉鸡创新团队建设项目（HBCT2018150203）

通讯作者: 赵国先,教授,博士生导师,E-mail:zgx959@163.com E-mail: zgx959@163.com

作者简介: 郭林霞(1992-),女,河北保定人,硕士研究生,研究方向为动物营养调控与畜产品品质。E-mail:1443050057@qq.com

引用本文:

郭林霞, 李树鹏, 马可为, 郝艳霜, 赵国先, 杨博文, 郑紫薇, 张达, 张思轩, 冯志华, 刘观忠. 枣多糖对小鼠脾脏B淋巴细胞体外增殖的影响[J]. 动物营养学报, 2020, 32(9): 4386-4392.
GUO Linxia, LI Shupeng, MA Kewei, HAO Yanshuang, ZHAO Guoxian, YANG Bowen, ZHENG Ziwei, ZHANG Da, ZHANG Sixuan, FENG Zhihua, LIU Guanzhong. Effects of Jujube Polysaccharide on Proliferation of Mouse Spleen B Lymphocytes in Vitro. Chinese Journal of Animal Nutrition, 2020, 32(9): 4386-4392.

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http://www.chinajan.com/CN/10.3969/j.issn.1006-267x.2020.09.049 或 http://www.chinajan.com/CN/Y2020/V32/I9/4386

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