太子参多糖对卵清白蛋白诱导免疫抑制小鼠肠道免疫应答影响的研究  

Effect of Pseudostellaria heterophylla polysaccharide on intestinal immune response in immunized mice induced by OVA

作　　者：曾靖棋 杜蓥蓥 周梦圆 曾丽 赵佳梅 陈颖清 熊太敏 李健[4] 马玉芳[1] ZENG Jing-qi;DU Ying-ying;ZHOU Meng-yuan;ZENG Li;ZHAO Jia-mei;CHEN Ying-qing;XIONG Tai-min;LI Jian;MA Yu-fang(University Key Laboratory for Integrated Chinese Traditional and Western Veterinary Medicine and Animal Healthcare in Fujian Province,College of Animal Sciences,Fujian Agriculture and Forestry University,Fuzhou 350002,China;Fujian Vocational College of Agriculture Technology,Fuzhou 350119,China;Institute of Animal Husbandry and Veterinary Medicine,Fujian Academy of Agriculture Sciences,Fuzhou 350013,China;Fujian Key Laboratory of Traditional Chinese Veterinary Medicine and Animal Health(Fujian Agriculture and Forestry University),Fuzhou 350002,China)

机构地区：[1]福建农林大学动物科学学院,中西兽医结合与动物保健重点实验室,福建福州350002 [2]福建农业职业技术学院,福建福州350119 [3]福建省农业科学院畜牧兽医研究所,福建福州350013 [4]福建省兽医中药与动物保健重点实验室(福建农林大学),福建福州350002

出　　处：《中国预防兽医学报》2024年第3期292-299,共8页Chinese Journal of Preventive Veterinary Medicine

基　　金：福建省重大专项专题项目(2021NZ02908)。

摘　　要：为研究太子参多糖(PHPS)对模式抗原卵清白蛋白(OVA)引起免疫抑制小鼠肠道免疫应答能力的影响,本研究将84只雄性ICR小鼠随机均分为7组,即PBS对照组(PBS组)、环磷酰胺对照组(CY组)、OVA单独免疫组(OVA-CY组)、铝佐剂对照组(OVA-CY-AL组)、OVA-CY-PHPS低、中、高剂量组(OVA-CY-PHPS-L组、OVACY-PHPS-M组、OVA-CY-PHPS-H组)。除PBS组以外,各组小鼠均于实验开始后1 d~3 d和18 d分别经腹腔注射环磷酰胺(CY,50 mg/kg),建立免疫抑制小鼠模型。于实验开始后4 d~8 d和19 d~23 d,OVA-CY-PHPS-L组、OVA-CY-PHPS-M组、OVA-CY-PHPS-H组小鼠分别灌胃对应剂量PHPS (50 mg/kg、100 mg/kg、200 mg/kg),PBS组、CY组、OVA-CY组和OVA-CY-AL组小鼠灌胃等体积双蒸水;第9 d和24 d,OVA-CY-PHPS-L组、OVA-CY-PHPS-M组、OVA-CY-PHPS-H组、OVA-CY组小鼠均经腹股沟皮下注射100μg OVA (0.2 m L/只)进行一免和二免。PBS组与CY组小鼠经相同方式注射等体积无菌PBS,OVA-AL组小鼠经相同方式注射OVA/铝佐剂混合液(OVA 100μg+Alum 200μg) 0.2 m L/只。第38 d剖杀各组小鼠,制作十二指肠、空肠、回肠病理切片观察小鼠肠道病变,采用Image-Pro Plus 6.0软件计算各肠段绒毛高度、隐窝深度、肠道上皮中的淋巴细胞数量;采用ELISA法检测各组小鼠各肠段IL-6、TNF-α含量、分泌型免疫球蛋白A (SIgA)抗体水平;采用荧光定量RT-PCR (qRT-PCR)法检测各组小鼠各肠段细胞因子m RNA的相对转录水平。结果显示,与OVA-CY组相比,各剂量PHPS均能改善免疫抑制小鼠肠道绒毛稀疏、肿胀、破裂的情况,低剂量的PHPS能够显著提高免疫抑制小鼠十二指肠、空肠绒毛高度与隐绒比(V/C)值(P<0.05),低、高剂量的PHPS均能够显著提高免疫抑制小鼠回肠的肠绒毛高度与V/C值(P<0.05);较OVA-CY组,高剂量PHPS能够显著提高免疫抑制小鼠空肠上皮内淋巴细胞的数量(P<0.05),低、高剂量PHPS能够显著提高免疫抑制小鼠回肠上皮内淋巴细胞的数量(To study the effect of Pseudostellaria heterophylla polysaccharide(PHPS) on the intestinal immune response of immunosuppressed mice induced by the model antigen ovalbumin(OVA),84 male ICR mice were randomly divided into 7 groups:PBS control group(PBS group),cyclophosphamide control group(CY group),OVA alone immunization group(OVA-CY group),aluminum adjuvant control group(OVA-CY-AL group),OVA-CY-PHPS low,medium,and high dose groups(OVA-CY-PHPS-L group,OVA-CY-PHPS-M group,OVA-CY-PHPS-H group).In order to establish immunosuppressed mouse models,excluding the PBS group,all the mice were intraperitoneally injected with cyclophosphamide(CY,50mg/kg) at 1st-3rd and 18th days.The mice in the OVA-CY-PHPS-L group,OVA-CY-PHPS-M group,and OVA-CY-PHPS-H group were given corresponding doses of PHPS(50mg/kg,100mg/kg,200mg/kg) by gavage at the 4th-8th and 19th-23rd days,while the other mice were given equal volumes of double distilled water by gavage in the PBS group,CY group,OVA-CY group,and OVA-CY-AL group;The first and second immunizations were performed by subcutaneously injecting 100μg OVA(0.2mL/animal) into the groin of mice in OVA-CY-PHPS-L group,OVA-CY-PHPS-M group,OVA-CY-PHPS-H group,and OVA-CY group at the 9th and 24th days.The mice in the PBS group and CY group were received subcutaneous injection with 0.2mL/animal of sterile PBS,while those in OVA AL group were received with 0.2mL/animal of OVA/aluminum adjuvant mixture into the groin(OVA 100μg+Alum 200μg).On the 38th day,the mice in each group were dissected and their tissue were made in pathological sections,and the intestinal lesions were observed in duodenum,jejunum,and ileum.Image Pro Plus 6.0 software was used to calculate the villus height,crypt depth,and lymphocyte count in the intestinal epithelium of each intestinal segment;The content of IL-6,TNF-α and immunoglobulin A(SIgA) antibody in various intestinal segments of mice in each group were detected by ELISA method;qRT-PCR was used to detect the relative transcription levels of cytokine m RNA in various

关 键 词：太子参多糖 免疫抑制小鼠 卵清白蛋白 肠道 免疫应答 

分 类 号：S852.6[农业科学—基础兽医学]