刺五加多糖纳米乳对小鼠的免疫调节和抗氧化作用

Immunomodulatory and antioxidant effects of Eleutherococcus senticosus polysaccharide nanoemulsion on mice

为了研究刺五加多糖纳米乳(Acanthopanax senticosus polysaccharide nanoemulsion,ASPS-NE)对小鼠的免疫调节和抗氧化作用,试验将70只ICR小鼠随机分成7组,分别为正常对照组、模型组、阳性组、刺五加多糖组及刺五加多糖纳米乳高、中、低剂量组,正常对照组每天按体重2 mL/kg皮下注射生理盐水,其余各组小鼠在试验第1,2,5天均按体重80 mg/kg皮下注射环磷酰胺建立免疫抑制模型;另外,阳性组每天按体重10 mg/kg皮下注射胸腺五肽,刺五加多糖组每天按体重62.5 mg/kg皮下注射刺五加多糖,刺五加多糖纳米乳高、中、低剂量组每天分别按体重125,62.5,31.25 mg/kg皮下注射刺五加多糖纳米乳,共7 d。末次给药24 h后处死小鼠后采集脾脏、心脏、肝脏及肾脏等脏器。分离脾细胞悬液,研究刺五加多糖纳米乳对小鼠脾淋巴细胞增殖的影响;采用实时荧光定量PCR方法检测腹腔巨噬细胞中白细胞介素-2(IL-2)、白细胞介素-4(IL-4)、γ干扰素(IFN-γ)、肿瘤坏死因子-α(TNF-α)mRNA的表达情况,并检测组织器官中谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)活性及丙二醛(MDA)浓度。结果表明:与模型组比较,刺五加多糖纳米乳3个剂量组的OD450值极显著升高(P<0.01),能不同程度促进小鼠脾淋巴细胞增殖,以刺五加多糖纳米乳中剂量组增殖效果明显;刺五加多糖纳米乳3个剂量组巨噬细胞中细胞因子IL-2、IL-4、IFN-γ和TNF-αmRNA的相对表达量极显著上调(P<0.01),以刺五加多糖纳米乳中剂量组细胞因子的分泌和相关mRNA的表达量更高;刺五加多糖纳米乳3个剂量组组织的GSH-Px、CAT活性极显著提高(P<0.01),并极显著降低组织的MDA浓度(P<0.01)。说明刺五加多糖纳米乳可增强免疫抑制小鼠的免疫调节和抗氧化作用。

In order to study the immunomodulatory and antioxidant effects of Eleutherococcus senticosus polysaccharide nanoemulsion(ASPSNE)in mice,in this experiment,seventy ICR mice were randomly divided into 7 groups,including normal control group,model group,positive group,ASPS group,and ASPS-NE high,medium and low dose groups.The normal control group was injected with normal saline subcutaneously according to the body weight of 2 mL/kg per day,and mice of the remaining groups were injected with 80 mg/kg subcutaneous cyclophosphamide on the 1st,2nd,and 5th days of the experiment to establish an immunosuppressive model.In addition,the positive group injected thymic pentapeptide subcutaneously at 10 mg/kg body weight per day.The ASPS group was injected subcutaneously with a body weight of 62.5 mg/kg per day.The high,medium and low dose groups of ASPS were injected subcutaneously with the body weight of 125,62.5 and 31.25 mg/kg per day respectively for a total of 7 days.24 h after the last dose,the mice were euthanized and the spleen,heart,liver and kidney were collected.The spleen cell suspension was isolated to study the effects of ASPS-NE on the proliferation of splenic lymphocytes in mice.The mRNA expression of interleukin-2(IL-2),interleukin-4(IL-4),γ-interferon(IFN-γ),tumor necrosis factor-α(TNF-α)in abdominal macrophages was detected by real-time PCR.The activity of glutathione peroxidase(GSH-Px),catalase(CAT),and malondialdehyde(MDA)concentration in tissues and organs were detected.The results showed that compared with the model group,the OD450 value of the three dose groups of ASPS-NE was significantly increased(P<0.01),and could promote the proliferation of splenic lymphocytes in mice to various degrees,and the proliferation effect of ASPS-NE medium group was obvious.The mRNA relative expression of cytokines IL-2,IL-4,IFN-γand TNF-αin macrophages was significantly upregulated in the three dose groups of ASPS-NE(P<0.01),and the secretion of cytokines and related mRNA expression in the ASPS-NE medium group were higher.The three dose groups of ASPS-NE significantly increased the activity of GSH-Px and CAT in tissues(P<0.01),and significantly reduced the concentration of MDA in tissues(P<0.01).The results suggested that ASPS-NE could enhance the immunomodulatory and antioxidant effects of immunosuppressed mice.