钝顶螺旋藻两个生态种多糖的免疫抗氧化功能的研究
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摘要
本论文以鄂尔多斯高原碱湖钝顶螺旋藻和非洲Chad湖钝顶螺旋藻为材料,制备并研究其多糖的免疫抗氧化活性,为筛选天然的抗氧化剂和免疫增强剂奠定理论研究依据。论文研究内容分为以下四部分:
(1)采用热水回流浸提法提取钝顶螺旋藻两个生态种多糖,三氯乙酸法去除蛋白并用活性碳进行脱色,得到鄂尔多斯高原碱湖钝顶螺旋藻多糖(S1多糖)和非洲Chad湖钝顶螺旋藻多糖(S2多糖)。
(2)采用比色法研究S1多糖和S2多糖体外清除超氧阴离子自由基(O2·-)和羟基自由基(·OH)的能力。结果显示:S1多糖和S2多糖在体外对自由基具有清除作用,且S1多糖的效果稍好于S2多糖。当多糖浓度为6 mg / mL时对O2·-的清除能力最强,分别为85.40 U / L、83.48 U / L;浓度为1.0 mg / mL时对·OH的清除能力最强,分别为32.56 U / mL、31.84 U / mL,且两种多糖体外对O2·-和·OH的清除能力在一定浓度范围内呈剂量依赖性,初步证实钝顶螺旋藻多糖具有抗氧化作用。
(3)采用试剂盒测定S1多糖和S2多糖的体内抗氧化能力。结果显示,各多糖组及藻泥组的抗氧化能力均高于空白对照组,且S1多糖和S2多糖的抗氧化能力要强于黄芪多糖对照组。①血清中,S1多糖灌胃浓度为1.25 mg / mL时,T-AOC含量最高(P <0.05),为45.78 U / mL,MDA含量最低,为7.73 nmol / mL;S2藻泥灌胃浓度为25 mg / mL时,CAT活力最高,为32.98 U / mL;S2多糖灌胃浓度为1.25 mg / mL时,GSH-PX活性最高,为2408.45 U / mL;S2多糖灌胃浓度为12.5 mg / mL时,T-SOD活性最高,为162.38 U / mL。②肝脏中,S2多糖灌胃浓度为1.25 mg / mL时,T-AOC含量最高,为3.74 U / mgprot、MDA含量最低,为2.57 nmol / mgprot;S2藻泥灌胃浓度为25 mg / mL时,CAT活力最高,为13.19 U / mgprot;S2多糖灌胃浓度为12.5 mg / mL时,GSH-PX活性最高,为518.78 U / mgprot;S2多糖灌胃浓度为1.25 mg / mL时,T-SOD活性最高,为159.44 U / mgprot。表明钝顶螺旋藻多糖能提高机体的抗氧化能力。
(4)采用试剂盒测定S1多糖和S2多糖的体内免疫作用。结果显示:各多糖组及藻泥组免疫能力均强于空白对照组,且S1多糖和S2多糖的免疫能力要强于黄芪多糖对照组。其中,S2多糖灌胃浓度为1.25 mg / mL时,胸腺与脾脏指数最高,分别为0.4188、0.4458,血清中ACP活力最高,为18.4867 U / 100mL,小鼠淋巴细胞转化率最高为47.5 %;S2多糖灌胃浓度为12.5 mg / mL时,肝脏指数最高为5.6708;S2藻泥灌胃浓度为25 mg / mL时,血清中溶菌酶含量最高,为8.02μg / mL;S1多糖灌胃浓度为1.25 mg / mL时,肝脏中ACP活力最高,为237.69 U / gprot。间接证明钝顶螺旋藻多糖具有提高机体免疫力的功效。
In this thesis, Polysaccharide was prepared from planus Spirulina.(Arthrospira) plantensis of the plateau natron lake Ordos and the Chard lake African, and it’s antioxidant activity of immune was studied. That laid a theoretical basis for further study. Thesis was divided into the following four parts:
(1)Two ecological types of polysaccharide were obtained with hot water extract . Protein was removed with trichloroacetic acid, and color was decolorized with activated carbon. Polysaccharide obtained from planus Spirulina.(Arthrospira) plantensis of the plateau natron lake Ordos named S1, and Polysaccharide obtained fromp lanus Spirulina.(Arthrospira) plantensis of the Chard lake African named S2.
(2) Capabilities were studied by colorimetry, that superoxide anion radical (O2 -) and hydroxyl radical (·OH) was scavenged by the polysaccharide. The results showed that: free radical could be scavenged by the polysaccharide(S1,S2) in vitro, and the effect of S1 was better than S2. free radical scavenging effect significantly. when the concentration of polysaccharide was 6 mg / mL , the capability of scavenging O2 - was 85.40 U / L and 83.48 U / L. when the contentration was 1.0 mg / mL, the effect was best on scavenging·OH, 32.56 U / mL and 31.84 U / mL. The effect existed in a dose-dependent on scavenging O2 - and·OH in a certain contentration range. Confirmed initially that polysaccharide from Spirulina platensis had antioxidant effects.
(3) The capacities of antioxidant in vivo were tested using the kits. Results showed that antioxidant capability of different doses of polysaccharide and algae mud treatment groups were higher than the control group, and antioxidant capability of polysaccharide were higher than that of the astragalus polysaccharides.①In the serum, the concentration of S1 polysaccharide was 1.25 mg / mL, the content of T-AOC was 45.78 U / mL (P <0.05), content of MDA was the lowest 7.73 nmol / mL. The concentration of algae mud S2 was 25 mg / mL, the activity of CAT was 32.98 U / mL. Concentration of polysaccharide S2 was1.25 mg / mL, the activity of GSH-PX was 2408.45 U / mL. The concentration of polysaccharide S2 was 12.5 mg / mL, the activity of T-SOD was 162.38 U / mL.②In the liver, the concentration of polysaccharide S2 was 1.25 mg / mL, the content of T-AOC was 3.74 U / mg prot, content of MDA was the lowest 2.57 nmol / mg prot. The concentration of algae mud S2 was 25 mg / mL, the activity of CAT was 13.19 U / mg prot. The concentration of polysaccharide S2 was 12.5 mg / mL, activity of GSH-PX was 518.78 U / mg prot. The concentration of polysaccharide S2 was 1.25 mg / mL, activity of T-SOD was 159.44 U / mg prot. Those showed that Spirulina platensis polysaccharide could enhance the body's antioxidant capacity.
(4)Immune function of the polysaccharide(S1,S2)was determined using kits. The results showed: immune capabilities of polysaccharide and algae mud treatments were higher than the control group, and immune capabilities of polysaccharides were higher than the astragalus polysaccharides. When the concentration of polysaccharide S2 was 1.25 mg / mL, the index of thymus and spleen were the 0.4188 and 0.4458 respectively, the activity of ACP was 18.4867 U/100mL in the serum, lymphocyte transformation rate was 47.5%. When the of concentration polysaccharide S2 was 12.5 mg / mL, the index of the liver was 5.6708. The concentration of algae mud S2 fed 25 mg / mL, content of lysozyme was 8.02μg / mL in the serum. concentration of polysaccharide S1 was 1.25 mg / mL, activity of ACP was 237.69 U / g prot in the liver. Those confirmed that Spirulina platensis polysaccharide could improve the immune function.
(1)采用热水回流浸提法提取钝顶螺旋藻两个生态种多糖,三氯乙酸法去除蛋白并用活性碳进行脱色,得到鄂尔多斯高原碱湖钝顶螺旋藻多糖(S1多糖)和非洲Chad湖钝顶螺旋藻多糖(S2多糖)。
(2)采用比色法研究S1多糖和S2多糖体外清除超氧阴离子自由基(O2·-)和羟基自由基(·OH)的能力。结果显示:S1多糖和S2多糖在体外对自由基具有清除作用,且S1多糖的效果稍好于S2多糖。当多糖浓度为6 mg / mL时对O2·-的清除能力最强,分别为85.40 U / L、83.48 U / L;浓度为1.0 mg / mL时对·OH的清除能力最强,分别为32.56 U / mL、31.84 U / mL,且两种多糖体外对O2·-和·OH的清除能力在一定浓度范围内呈剂量依赖性,初步证实钝顶螺旋藻多糖具有抗氧化作用。
(3)采用试剂盒测定S1多糖和S2多糖的体内抗氧化能力。结果显示,各多糖组及藻泥组的抗氧化能力均高于空白对照组,且S1多糖和S2多糖的抗氧化能力要强于黄芪多糖对照组。①血清中,S1多糖灌胃浓度为1.25 mg / mL时,T-AOC含量最高(P <0.05),为45.78 U / mL,MDA含量最低,为7.73 nmol / mL;S2藻泥灌胃浓度为25 mg / mL时,CAT活力最高,为32.98 U / mL;S2多糖灌胃浓度为1.25 mg / mL时,GSH-PX活性最高,为2408.45 U / mL;S2多糖灌胃浓度为12.5 mg / mL时,T-SOD活性最高,为162.38 U / mL。②肝脏中,S2多糖灌胃浓度为1.25 mg / mL时,T-AOC含量最高,为3.74 U / mgprot、MDA含量最低,为2.57 nmol / mgprot;S2藻泥灌胃浓度为25 mg / mL时,CAT活力最高,为13.19 U / mgprot;S2多糖灌胃浓度为12.5 mg / mL时,GSH-PX活性最高,为518.78 U / mgprot;S2多糖灌胃浓度为1.25 mg / mL时,T-SOD活性最高,为159.44 U / mgprot。表明钝顶螺旋藻多糖能提高机体的抗氧化能力。
(4)采用试剂盒测定S1多糖和S2多糖的体内免疫作用。结果显示:各多糖组及藻泥组免疫能力均强于空白对照组,且S1多糖和S2多糖的免疫能力要强于黄芪多糖对照组。其中,S2多糖灌胃浓度为1.25 mg / mL时,胸腺与脾脏指数最高,分别为0.4188、0.4458,血清中ACP活力最高,为18.4867 U / 100mL,小鼠淋巴细胞转化率最高为47.5 %;S2多糖灌胃浓度为12.5 mg / mL时,肝脏指数最高为5.6708;S2藻泥灌胃浓度为25 mg / mL时,血清中溶菌酶含量最高,为8.02μg / mL;S1多糖灌胃浓度为1.25 mg / mL时,肝脏中ACP活力最高,为237.69 U / gprot。间接证明钝顶螺旋藻多糖具有提高机体免疫力的功效。
In this thesis, Polysaccharide was prepared from planus Spirulina.(Arthrospira) plantensis of the plateau natron lake Ordos and the Chard lake African, and it’s antioxidant activity of immune was studied. That laid a theoretical basis for further study. Thesis was divided into the following four parts:
(1)Two ecological types of polysaccharide were obtained with hot water extract . Protein was removed with trichloroacetic acid, and color was decolorized with activated carbon. Polysaccharide obtained from planus Spirulina.(Arthrospira) plantensis of the plateau natron lake Ordos named S1, and Polysaccharide obtained fromp lanus Spirulina.(Arthrospira) plantensis of the Chard lake African named S2.
(2) Capabilities were studied by colorimetry, that superoxide anion radical (O2 -) and hydroxyl radical (·OH) was scavenged by the polysaccharide. The results showed that: free radical could be scavenged by the polysaccharide(S1,S2) in vitro, and the effect of S1 was better than S2. free radical scavenging effect significantly. when the concentration of polysaccharide was 6 mg / mL , the capability of scavenging O2 - was 85.40 U / L and 83.48 U / L. when the contentration was 1.0 mg / mL, the effect was best on scavenging·OH, 32.56 U / mL and 31.84 U / mL. The effect existed in a dose-dependent on scavenging O2 - and·OH in a certain contentration range. Confirmed initially that polysaccharide from Spirulina platensis had antioxidant effects.
(3) The capacities of antioxidant in vivo were tested using the kits. Results showed that antioxidant capability of different doses of polysaccharide and algae mud treatment groups were higher than the control group, and antioxidant capability of polysaccharide were higher than that of the astragalus polysaccharides.①In the serum, the concentration of S1 polysaccharide was 1.25 mg / mL, the content of T-AOC was 45.78 U / mL (P <0.05), content of MDA was the lowest 7.73 nmol / mL. The concentration of algae mud S2 was 25 mg / mL, the activity of CAT was 32.98 U / mL. Concentration of polysaccharide S2 was1.25 mg / mL, the activity of GSH-PX was 2408.45 U / mL. The concentration of polysaccharide S2 was 12.5 mg / mL, the activity of T-SOD was 162.38 U / mL.②In the liver, the concentration of polysaccharide S2 was 1.25 mg / mL, the content of T-AOC was 3.74 U / mg prot, content of MDA was the lowest 2.57 nmol / mg prot. The concentration of algae mud S2 was 25 mg / mL, the activity of CAT was 13.19 U / mg prot. The concentration of polysaccharide S2 was 12.5 mg / mL, activity of GSH-PX was 518.78 U / mg prot. The concentration of polysaccharide S2 was 1.25 mg / mL, activity of T-SOD was 159.44 U / mg prot. Those showed that Spirulina platensis polysaccharide could enhance the body's antioxidant capacity.
(4)Immune function of the polysaccharide(S1,S2)was determined using kits. The results showed: immune capabilities of polysaccharide and algae mud treatments were higher than the control group, and immune capabilities of polysaccharides were higher than the astragalus polysaccharides. When the concentration of polysaccharide S2 was 1.25 mg / mL, the index of thymus and spleen were the 0.4188 and 0.4458 respectively, the activity of ACP was 18.4867 U/100mL in the serum, lymphocyte transformation rate was 47.5%. When the of concentration polysaccharide S2 was 12.5 mg / mL, the index of the liver was 5.6708. The concentration of algae mud S2 fed 25 mg / mL, content of lysozyme was 8.02μg / mL in the serum. concentration of polysaccharide S1 was 1.25 mg / mL, activity of ACP was 237.69 U / g prot in the liver. Those confirmed that Spirulina platensis polysaccharide could improve the immune function.
引文
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