骆驼刺酸性多糖对小鼠脾脏淋巴细胞免疫增强作用的研究  

Study on the immune enhancement effect of acidic Alhagi sparsifolia Shap polysaccharide on splenic lymphocytes of mice

作　　者：特列克·阿依恒别克 赛福丁·阿不拉[1] 萨比热·热夏提 况玲[1] 德力拜尔·木拉提 阿得力江·吾斯曼 TELIEKE·Ayihengbieke;SAIFUDING·Abula;SABIRE·Rexiati;KUANG Ling;DILIBAIER·Mulati;ADELIJIANG·Wusiman(College of Veterinary Medicine,Xinjiang Agricultural University,Urumqi 830052,China;Xinjiang Uigar Autonomous Regional Center for Animal Disease Control and Prevention,Urumqi 830011,China)

机构地区：[1]新疆农业大学动物医学学院,新疆乌鲁木齐830052 [2]新疆维吾尔自治区动物疫病预防控制中心,新疆乌鲁木齐830011

出　　处：《南京农业大学学报》2022年第6期1220-1226,共7页Journal of Nanjing Agricultural University

基　　金：中国博士后科学基金项目(2021M693899);新疆维吾尔自治区自然科学计划(自然科学基金)青年科学基金项目(2021D01B48);青年博士科技人才培养计划-天山青年项目(2021Q076)。

摘　　要：[目的]本文旨在探讨骆驼刺酸性多糖(acidic Alhagi sparsifolia Shap polysaccharide,aASP)对小鼠脾淋巴细胞的免疫增强作用。[方法]采用醇沉法提取骆驼刺多糖,采用柱层析法对骆驼刺多糖进行分离及纯化,采用苯酚硫酸法、硫酸-咔唑法和BCA微孔酶标仪法,检测糖、糖醛酸和蛋白含量,采用红外光谱法鉴别多糖性质。采用MTT法确定aASP对小鼠脾脏淋巴细胞的安全浓度,在安全浓度范围内500、125和31.25μg·mL^(-1)3个浓度下检测单独、协同脂多糖(lipopolysaccharide,LPS)和植物血凝素(phytoheamagglutinin,PHA)对脾淋巴细胞的增殖效果;流式细胞术检测aASP对淋巴细胞亚群分化的影响;ELISA法检测aASP对淋巴细胞细胞因子分泌的影响。[结果]骆驼刺多糖提取物中多糖含量为(81.8±1.2)%,糖醛酸含量为(17.34±0.75)%,蛋白含量为(0.54±0.08)%;红外光谱显示aASP在1746.53 cm^(-1)处有较强的吸收峰,结合糖醛酸含量判定aASP为酸性多糖。高、中、低浓度的aASP均可促进脾脏淋巴细胞增殖,协同LPS和PHA能够更显著促进B细胞和T淋巴细胞的增殖(P<0.05)。流式细胞术和ELISA检测结果显示aASP能够促进CD4^(+)和CD8a^(+)T细胞的分化,并促进脾淋巴细胞分泌IFN-γ、TNF-α、IL-6和IL-4。[结论]aASP可以增强小鼠脾脏淋巴细胞的增殖,诱导小鼠脾脏T淋巴细胞产生Th1(helper T lymphocyte 1)及Th2(helper T lymphocyte 2)混合型免疫应答,免疫细胞偏向CD4^(+)T细胞分化。[Objectives]The paper aimed to investigate the immunological enhancement of acidic Alhagi sparsifolia Shap polysaccharide(aASP)on mouse splenic lymphocytes.[Methods]aASP was extracted by alcohol precipitation,separated and purified by column chromatography,and the contents of sugar,uronic acid,and protein were detected using phenol sulfuric acid method,sulfuric acid-carbazole method,and BCA microplate reader.The polysaccharide properties were identified by infrared spectroscopy.MTT assay was used to determine the safe concentration of aASP on mouse splenic lymphocytes,and the proliferative effects of aASP alone and synergistic lipopolysaccharide(LPS)and phytoheamagglutinin(PHA)on splenic lymphocytes were detected at three concentrations of 500,125 and 31.25μg·mL^(-1)within the safe concentration range;the effect of aASP on lymphocyte differentiation was detected by flow cytometry;the effect of aASP on lymphocyte cytokine secretion was detected by ELISA.[Results]The contents of polysaccharide,uronic acid,and protein in aASP extracts were(81.8±1.2)%,(17.34±0.75)%,and(0.54±0.08)%,respectively.Infrared spectra showed that aASP had a strong absorption peak at 1746.53 cm^(-1),and combined with the content of uronic acid,aASP could be identified as acidic polysaccharide.High,medium,and low concentrations of aASP could promote the proliferation of splenic lymphocytes,and aASP synergistic with LPS and PHA could significantly promote the proliferation of B cells and T lymphocytes(P<0.05).The results of flow cytometry and ELISA showed that aASP was able to promote the differentiation of CD4^(+)and CD8a^(+)T cells and promote the secretion of IFN-γ,TNF-α,IL-6,and IL-4 in splenic lymphocytes.[Conclusions]aASP can boost the proliferation of splenic lymphocytes in mice,induce Th1(helper T lymphocyte 1)and Th2(helper T lymphocyte 2)mixed immune responses in splenic T lymphocytes,and the immune cells are biased towards CD4^(+)T cell differentiation.

关 键 词：骆驼刺酸性多糖 脾脏淋巴细胞 免疫增强 细胞分化 

分 类 号：S853.71[农业科学—临床兽医学]