EPSAH is an exopolysaccharide from Aphanothece halophytica GR02. The present study was designed to evaluate its toxicity and adjuvant potential in the specific cellular and humoral immune responses in ovalbumin(OVA) i...EPSAH is an exopolysaccharide from Aphanothece halophytica GR02. The present study was designed to evaluate its toxicity and adjuvant potential in the specific cellular and humoral immune responses in ovalbumin(OVA) in mice. EPSAH did not cause any mortality and side effects when the mice were administered subcutaneously twice at the dose of 50 mg×kg^(–1). Hemolytic activity in vitro indicated that EPSAH was non-hemolytic. Splenocyte proliferation in vitro was assayed with different concentrations of EPSAH. The mice were immunized subcutaneously with OVA 0.1 mg alone or with OVA 0.1 mg dissolved in saline containing Alum(0.2 mg) or EPSAH(0.2, 0.4, or 0.8 mg) on Day 1 and 15. Two weeks later, splenocyte proliferation, natural killer(NK) cell activity, production of cytokines IL-2 from splenocytes, and serum OVA-specific antibody titers were measured. Phagocytic activity, production of pro-inflammatory cytokines IL-1 and IL-12 in mice peritoneal macrophages were also determined. EPSAH showed a dose-dependent stimulating effect on mitogen-induced proliferation. The Con A-, LPS-, and OVA-induced splenocyte proliferation and the serum OVA-specific IgG, IgG1, and IgG2 a antibody titers in the immunized mice were significantly enhanced. EPSAH also significantly promoted the production of Th1 cytokine IL-2. Besides, EPSAH remarkably increased the killing activities of NK cells from splenocytes in the immunized mice. In addition, EPSAH enhanced phagocytic activity and the generation of pro-inflammatory cytokines IL-1 and IL-12 in macrophages. These results indicated that EPSAH had a strong potential to increase both cellular and humoral immune responses, particularly promoting the development of Th1 polarization.展开更多
目的探讨嗜盐隐杆藻胞外多糖(exopo lysaccharide of Aphanothece ha lophy tica,EPAH)的抗肿瘤活性。方法以S180肉瘤为肿瘤模型,检测EPAH对肿瘤生长的抑制活性;M TT法检测EPAH对S180肉瘤细胞、Sm cc7721肝癌细胞和H eL a细胞的体外抑...目的探讨嗜盐隐杆藻胞外多糖(exopo lysaccharide of Aphanothece ha lophy tica,EPAH)的抗肿瘤活性。方法以S180肉瘤为肿瘤模型,检测EPAH对肿瘤生长的抑制活性;M TT法检测EPAH对S180肉瘤细胞、Sm cc7721肝癌细胞和H eL a细胞的体外抑制活性;以对荷瘤小鼠免疫器官、血液淋巴细胞数量及淋巴细胞增殖影响、NK细胞杀伤活性和巨噬细胞产生NO及其他细胞因子的影响评价EPAH对小鼠免疫功能的作用。结果EPAH对S180肉瘤生长有显著的抑制作用,其中预防给药和与肿瘤接种同时给药组最大抑瘤率分别达66.79%和47.93%。100μg/mL EPAH对Sm cc7721细胞、H eL a细胞以及S180细胞生长的抑制率达60%以上。EPAH显著提高荷瘤小鼠脾脏和胸腺质量以及血液淋巴细胞数量,促进淋巴细胞增殖反应,增加NK细胞杀伤活性,刺激巨噬细胞产生NO,分泌白细胞介素-1β(IL-1β)及肿瘤坏死因子-α(TNF-α)。结论EPAH有显著的抗肿瘤活性,并能直接作用于肿瘤细胞,抑制肿瘤生长。EPAH可能是通过增加荷瘤小鼠免疫器官的质量和免疫细胞的数量、促进淋巴细胞分裂,提高NK细胞杀伤活性和巨噬细胞产生NO以及其他细胞因子实现其抗肿瘤活性。展开更多
The effects of the varying concentrations of the polysaccharide, eletrolyte and different pH values and temperature on the viscosity of the exopolysaccharide produced by \%Aphanothece halophytica \%Fremy were analysed...The effects of the varying concentrations of the polysaccharide, eletrolyte and different pH values and temperature on the viscosity of the exopolysaccharide produced by \%Aphanothece halophytica \%Fremy were analysed. The results showed that its intrinsic viscosity is 9.76 dl/g, which provides good stability to salt, pH and temperature.展开更多
文章确立了简便的一步法纯化盐生隐杆藻藻蓝蛋白。经过磷酸缓冲液提取、硫酸铵分步沉淀和DEAE-Sepharose Fast F low柱层析,分离纯化得到盐生隐杆藻藻蓝蛋白,其纯度(A620/A280)达到4.58,最终得率为52.65%。纯化的藻蓝蛋白最大吸收峰为62...文章确立了简便的一步法纯化盐生隐杆藻藻蓝蛋白。经过磷酸缓冲液提取、硫酸铵分步沉淀和DEAE-Sepharose Fast F low柱层析,分离纯化得到盐生隐杆藻藻蓝蛋白,其纯度(A620/A280)达到4.58,最终得率为52.65%。纯化的藻蓝蛋白最大吸收峰为620nm,其室温荧光发射峰为646nm,SDS-PAGE电泳显示其α和β亚基的分子量分别为17218Da和20844Da,分子排阻HPLC测得其分子量为76158Da,表明其在pH 7.2下是以(αβ)2二聚体状态存在于溶液中。稳定性实验表明,黑暗、pH 5~8以及0M^2M NaC l等条件下AH-PC是稳定的。这些实验结果将为开发利用盐生隐杆藻藻蓝蛋白提供参考。展开更多
基金supported by College Students Innovation Project for the R&D of Novel Drugs(J1030830)Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)
文摘EPSAH is an exopolysaccharide from Aphanothece halophytica GR02. The present study was designed to evaluate its toxicity and adjuvant potential in the specific cellular and humoral immune responses in ovalbumin(OVA) in mice. EPSAH did not cause any mortality and side effects when the mice were administered subcutaneously twice at the dose of 50 mg×kg^(–1). Hemolytic activity in vitro indicated that EPSAH was non-hemolytic. Splenocyte proliferation in vitro was assayed with different concentrations of EPSAH. The mice were immunized subcutaneously with OVA 0.1 mg alone or with OVA 0.1 mg dissolved in saline containing Alum(0.2 mg) or EPSAH(0.2, 0.4, or 0.8 mg) on Day 1 and 15. Two weeks later, splenocyte proliferation, natural killer(NK) cell activity, production of cytokines IL-2 from splenocytes, and serum OVA-specific antibody titers were measured. Phagocytic activity, production of pro-inflammatory cytokines IL-1 and IL-12 in mice peritoneal macrophages were also determined. EPSAH showed a dose-dependent stimulating effect on mitogen-induced proliferation. The Con A-, LPS-, and OVA-induced splenocyte proliferation and the serum OVA-specific IgG, IgG1, and IgG2 a antibody titers in the immunized mice were significantly enhanced. EPSAH also significantly promoted the production of Th1 cytokine IL-2. Besides, EPSAH remarkably increased the killing activities of NK cells from splenocytes in the immunized mice. In addition, EPSAH enhanced phagocytic activity and the generation of pro-inflammatory cytokines IL-1 and IL-12 in macrophages. These results indicated that EPSAH had a strong potential to increase both cellular and humoral immune responses, particularly promoting the development of Th1 polarization.
文摘目的探讨嗜盐隐杆藻胞外多糖(exopo lysaccharide of Aphanothece ha lophy tica,EPAH)的抗肿瘤活性。方法以S180肉瘤为肿瘤模型,检测EPAH对肿瘤生长的抑制活性;M TT法检测EPAH对S180肉瘤细胞、Sm cc7721肝癌细胞和H eL a细胞的体外抑制活性;以对荷瘤小鼠免疫器官、血液淋巴细胞数量及淋巴细胞增殖影响、NK细胞杀伤活性和巨噬细胞产生NO及其他细胞因子的影响评价EPAH对小鼠免疫功能的作用。结果EPAH对S180肉瘤生长有显著的抑制作用,其中预防给药和与肿瘤接种同时给药组最大抑瘤率分别达66.79%和47.93%。100μg/mL EPAH对Sm cc7721细胞、H eL a细胞以及S180细胞生长的抑制率达60%以上。EPAH显著提高荷瘤小鼠脾脏和胸腺质量以及血液淋巴细胞数量,促进淋巴细胞增殖反应,增加NK细胞杀伤活性,刺激巨噬细胞产生NO,分泌白细胞介素-1β(IL-1β)及肿瘤坏死因子-α(TNF-α)。结论EPAH有显著的抗肿瘤活性,并能直接作用于肿瘤细胞,抑制肿瘤生长。EPAH可能是通过增加荷瘤小鼠免疫器官的质量和免疫细胞的数量、促进淋巴细胞分裂,提高NK细胞杀伤活性和巨噬细胞产生NO以及其他细胞因子实现其抗肿瘤活性。
文摘The effects of the varying concentrations of the polysaccharide, eletrolyte and different pH values and temperature on the viscosity of the exopolysaccharide produced by \%Aphanothece halophytica \%Fremy were analysed. The results showed that its intrinsic viscosity is 9.76 dl/g, which provides good stability to salt, pH and temperature.
文摘文章确立了简便的一步法纯化盐生隐杆藻藻蓝蛋白。经过磷酸缓冲液提取、硫酸铵分步沉淀和DEAE-Sepharose Fast F low柱层析,分离纯化得到盐生隐杆藻藻蓝蛋白,其纯度(A620/A280)达到4.58,最终得率为52.65%。纯化的藻蓝蛋白最大吸收峰为620nm,其室温荧光发射峰为646nm,SDS-PAGE电泳显示其α和β亚基的分子量分别为17218Da和20844Da,分子排阻HPLC测得其分子量为76158Da,表明其在pH 7.2下是以(αβ)2二聚体状态存在于溶液中。稳定性实验表明,黑暗、pH 5~8以及0M^2M NaC l等条件下AH-PC是稳定的。这些实验结果将为开发利用盐生隐杆藻藻蓝蛋白提供参考。
