新疆栽培荒漠肉苁蓉醇提物的免疫调节作用研究

Immunomodulatory activity of ethanol extracts of cultivated Cistanche deserticola in Xinjiang

目的探讨新疆栽培荒漠肉苁蓉醇提物(EECCD)的免疫调节活性。方法以卵清蛋白(OVA)为抗原,将ICR小鼠分为9 g/L NaCl组(空白对照组)、EECCD组(1 200 μg EECCD)、OVA组(10 μg OVA)、低剂量EECCD/OVA组(400 μg EECCD+10 μg OVA)、中剂量EECCD/OVA组(800 μg EECCD+10 μg OVA)、高剂量EECCD/OVA组(1 200 μg EECCD+10 μg OVA)和铝佐剂(Alum)/OVA组(200 μg Alum+10 μg OVA)。皮下免疫小鼠,初免14 d后加强免疫1次。采用间接酶联免疫吸附测定法检测OVA特异性抗体水平,噻唑蓝法检测脾淋巴细胞增殖水平,流式细胞术检测白细胞介素-4(IL-4)和γ-干扰素(IFN-γ)水平,并检测EECCD对树突状细胞(DCs)共刺激分子的表达和CD4+CD25+Foxp3+调节性T细胞的影响。结果 EECCD可提高抗体水平,具体表现为:低、中、高剂量EECCD/OVA均提高了OVA特异性IgG抗体滴度,中剂量EECCD/OVA组抗体滴度达25万,与Alum/OVA组相当;中剂量EECCD/OVA明显提高了OVA特异性IgG1水平(P<0.01),且与Alum/OVA组相比差异无统计学意义(P>0.05);中剂量和高剂量EECCD/OVA明显提高了IgG2a水平(均P<0.01),且中剂量EECCD/OVA组明显高于Alum/OVA组(P<0.01)。由此筛选出中剂量EECCD为最佳剂量。中剂量EECCD/OVA能明显提高脾淋巴细胞的增殖水平(P<0.05),促进CD4+ T细胞分泌IL-4和IFN-γ(均P<0.01),还可明显提高DCs表面分子(CD40、CD80、CD86、MHCⅡ)的表达(均P<0.05),降低CD4+CD25+Foxp3+调节性T细胞的表达(P<0.05)。结论 EECCD具有良好的免疫调节活性,能促进偏向Th1型免疫应答,对疾病的预防和治疗具有潜在的作用。

Objective To explore the immunomodulatory activity of ethanol extract of cultivated Cistanche deserticola(EECCD)in Xinjiang.Methods Ovalbumin(OVA)was used antigen,ICR mice were divided into 9 g/L NaCl group(blank control group),EECCD group(1200μg EECCD),OVA group(10μg OVA),low-dose EECCD/OVA group(400μg EECCD+10μg OVA),medium-dose EECCD/OVA group(800μg EECCD+10μg OVA),high-dose EECCD/OVA group(1200μg EECCD+10μg OVA)and aluminum adjuvant(Alum)/OVA group(200μg Alum+10μg OVA).Mice were immunized subcutaneously,and the immunization was strengthened once 14 days after the initial immunization.The level of splenocyte proliferation was determined by thiazolyl blue tetrazolium bromide(MTT)method,and interferonγ(IFN-γ)and interleukin-4(IL-4)in CD4+T cell,dendritic cells(DCs)surface markers and CD4+CD25+Foxp3+Treg were evaluated by fluorescence-activated cell sorting(FACS).Results Three dose of EECCD can enhance OVA-specific IgG titers in serum.The antibody titer in medium-dose EECCD/OVA group was 250000,which was the same as that in the Alum/OVA group.The medium-dose EECCD/OVA significantly improve IgG1 and IgG2a(both P<0.01).Therefore,the medium dose EECCD was selected as the best dose.MTT results displayed that splenocyte proliferation were significantly stimulated by medium-dose EECCD/OVA(P<0.05),and the levels of IL-4 and IFN-γin CD4+T cells were promoted in groups administered with medium-dose EECCD/OVA(both P<0.01).Furthermore,medium-dose EECCD/OVA significantly up-regulated the levels of CD40,CD80,CD86 and major histocompatibility complex classⅡ(MHCⅡ)on DCs and down-regulated the frequency of CD4+CD25+Foxp3+Treg(all P<0.05).Conclusions EECCD has good immunomodulatory activity,can promote Th1-biased response,and has the therapeutic potential for the prevention of diseases.