抗菌肽Sublancin与黄芪多糖对小鼠免疫调节作用的比较研究

Comparative Study on Immunomodulatory Effect of Antimicrobial Peptide Sublancin and Astragalus Polysaccharide of Mice

本试验旨在比较抗菌肽Sublancin与黄芪多糖对小鼠免疫功能的调节作用。选用4~6周龄健康雌性BALB/c小鼠60只,随机分成6组,每组10只。各组小鼠先连续灌胃7 d、每天1次、每只0.2 m L的下列物质:空白对照组,灌胃生理盐水;攻毒组,灌胃生理盐水;12.0 mg/kg BW黄芪多糖组,灌胃12.0 mg/kg BW的黄芪多糖溶液;48.0 mg/kg BW黄芪多糖组,灌胃48.0 mg/kg BW的黄芪多糖溶液;1.0 mg/kg BW Sublancin组,灌胃1.0 mg/kg BW的Sublancin溶液;2.0 mg/kg BW Sublancin组,灌胃2.0 mg/kg BW的Sublancin溶液。除空白对照组外,其余5组在灌胃结束24 h后均以200μL/只的剂量灌胃浓度为1×109CFU/m L的鼠伤寒沙门氏菌进行攻毒。分别在攻毒3和24 h后从每组随机取5只小鼠采集外周血、脾脏以及盲肠内容物,检测血清中细胞因子含量,脾细胞中T淋巴细胞亚群CD+4、CD_8^+数量以及肠道内容物中沙门氏菌数量等指标。结果表明:沙门氏菌攻毒3 h后,与攻毒组相比,12.0和48.0 mg/kg BW黄芪多糖组以及2.0 mg/kg BW Sublancin组血清中白细胞介素-6(IL-6)的含量均显著降低(P<0.05);1.0 mg/kg BW Sublancin组血清中白细胞介素-10(IL-10)的含量显著提高(P<0.05);12.0和48.0 mg/kg BW黄芪多糖组以及1.0和2.0 mg/kg BW Sublancin组血清中肿瘤坏死因子-α(TNF-α)的含量均显著降低(P<0.05),但血清中白细胞介素-25(IL-25)的含量无显著变化(P>0.05);48.0 mg/kg BW黄芪多糖组和2.0 mg/kg BW Sublancin组血清中单核细胞趋化蛋白-1(MCP-1)的含量均显著降低(P<0.05);48.0 mg/kg BW黄芪多糖组以及1.0和2.0 mg/kg BW Sublancin组脾细胞中CD+4/CD_8^+均显著提高(P<0.05);12.0和48.0 mg/kg BW黄芪多糖组以及1.0和2.0 mg/kg BW Sublancin组肠道内容物中沙门氏菌的数量均没有产生显著变化(P>0.05),但是存在降低趋势。沙门氏菌攻毒24 h后,与攻毒组相比,2.0 mg/kg BW Sublancin组血清中IL-6的含量显著降低(P<0.05);48.0 mg/kg BW黄芪多糖组和2.0 mg/kg BW Sublancin组血清中TNF-α的含量显著降低(P<0.05);12.0和48.0 mg/kg BW黄芪多糖组以及2.0 mg/kg BW Sublancin组血清中MCP-1的含量显著降低(P<0.05)。此外,沙门氏菌攻毒24 h后,与空白对照组相比,2.0和48.0 mg/kg BW黄芪多糖组以及1.0和2.0 mg/kg BW Sublancin组血清中IL-10的含量均显著提高(P<0.05)。综合上述结果可得出,适宜剂量的抗菌肽Sublancin和黄芪多糖对小鼠免疫功能均有良好的调节作用;与黄芪多糖相比,抗菌肽Sublancin对感染沙门氏菌小鼠免疫功能的调节更加全面。

This experiment was conducted to compare the regulating effect of antimicrobial peptide Sublancin and Astragalus polysaccharide on immune function of mice.Sixty female BALB/c mice at the age of 4 to 6 weeks were randomly assigned into 6 groups with 10 mice per group.Mice in blank control group received sa-line by intragastric administration,mice in challenged group received saline by intragastric administration,mice in 12.0 and 48.0 mg/kg BW Astragalus polysaccharide groups received 12.0 and 48.0 mg/kg BW Astragalus polysaccharide solution by intragastric administration,respectively,and mice in 1.0 and 2.0 mg/kg BW Sub-lancin groups received 1.0 and 2.0 mg/kg BW Sublancin solution by intragastric administration,respectively.All the mice were administrated orally daily for 7 days,and the dose of each mouse was 0.2 mL.Except blank control group,mice in other 5 groups were challenged with Salmonella typhimurium(1×109 CFU/mL,200μL per mouse)on day 8(24 h after intragastric administration).Five mice from each group were ran-domly selected 3 and 24 h after challenge and the peripheral blood,spleen and caecum contents were sampled to measure the serum cytokine contents,T lymphocyte subsets CD+4 and CD+8 numbers in splenocyte and Salmo-nella number in intestinal contents,and so on.The results showed as follows:challenged 3 h by Salmonella,compared with the challenged group,the serum interleukin-6(IL-6)content in 12.0 and 48.0 mg/kg BW As-tragalus polysaccharide groups and 2.0 mg/kg BW Sublancin group was significantly reduced(P<0.05);the serum interleukin-10(IL-10)content in 1.0 mg/kg BW Sublancin group was significantly increased(P<0.05);the serum tumor necrosis factor-α(TNF-α)content in 12.0 and 48.0 mg/kg BW Astragalus polysac-charide groups,1.0 and 2.0 mg/kg BW Sublancin groups was significantly reduced(P<0.05),but the serum interleukin-25(IL-25)content was not significantly different(P>0.05);the serum monocyte chemoattractant protein-1(MCP-1)content in 48.0 mg/kg BW Astragalus polysaccharide group and 2.0 mg/kg BW Sublancin group was significantly reduced(P<0.05);CD+4/CD+8 in splenocyte in 48.0 mg/kg BW Astragalus polysaccharide group,1.0 and 2.0 mg/kg BW Sublancin groups was significantly increased(P<0.05);the Salmonel-la number in intestinal contents in 12.0 and 48.0 mg/kg BW Astragalus polysaccharide groups,1.0 and 2.0 mg/kg BW Sublancin groups had no significant changes(P>0.05),but had reducing trend.Challenged 24 h by Salmonella,compared with the challenged group,the serum IL-6 content in 2.0 mg/kg BW Sublancin group was significantly reduced(P<0.05);the serum TNF-αcontent in 48.0 mg/kg BW Astragalus polysac-charide group and 2.0 mg/kg BW Sublancin group was significantly reduced(P<0.05);the serum MCP-1 content in 12.0 and 48.0 mg/kg BW Astragalus polysaccharide groups,2.0 mg/kg BW Sublancin group was significantly reduced(P<0.05).Moreover,the serum IL-10 content in 12.0 and 48.0 mg/kg BW Astragalus polysaccharide groups,1.0 and 2.0 mg/kg BW Sublancin groups was significantly increased challenged 24 h by Salmonella compared with blank control group(P<0.05).These results indicate that antimicrobial peptide Sublancin and Astragalus polysaccharide have regulating effect on immune function for mice,and the immuno-modulatory effect of antimicrobial peptide Sublancin is more comprehensive than that of Astragalus polysaccha-ride.