摘要
试验旨在研究刺五加多糖(ASPS)对免疫抑制雏鸡脾脏中T淋巴细胞分布及免疫功能的影响,从而对ASPS的免疫增强作用进行评价。300只4日龄海兰褐公雏随机分为3组,每组100只:第1组为空白对照组,第2组为环磷酰胺(CTX)致免疫抑制组,第3组为ASPS+CTX组。第2组和第3组按照80 mg/kg·d的剂量皮下注射CTX,连续注射3 d。从10日龄开始,第3组每只鸡每天皮下注射0.2 m L 200 mg/m L的ASPS,连续注射3 d。第1组和第2组以同样方式注射等量灭菌生理盐水。注射后的第7、14、21和28天分别取脾脏,通过免疫组织化学染色和流式细胞术检测脾脏中CD4+、CD8+T淋巴细胞分布和数量变化,并利用荧光定量PCR方法检测脾脏中IFN-γ和IL-2 m RNA表达水平的变化。结果显示:CTX能够造成脾脏中CD4+和CD8+T淋巴细胞数量均显著减少(P<0.05),主要由CD4+细胞构成的PALS数量减少或消失,红髓和白髓中的CD4+和CD8+T淋巴细胞分布范围均减小,而且细胞排列疏松,IFN-γ和IL-2m RNA表达水平较空白对照组显著减少(P<0.05);在ASPS的作用下,ASPS+CTX组CD4+和CD8+T淋巴细胞的数量均比CTX组显著增加(P<0.05),PALS面积逐渐增加,而且结构逐渐完整清晰,CD8+T淋巴细胞的分布范围显著增加,红髓和白髓中CD8+细胞分布更加密集,免疫后28 d时基本恢复正常水平,IFN-γ和IL-2 m RNA表达水平显著提高(P<0.05)。结果表明,ASPS对CTX造成的脾脏中CD4+和CD8+T淋巴细胞定位分布和数量异常以及IFN-γ和IL-2 m RNA表达水平下降有显著的修复作用,证明ASPS对鸡脾脏免疫功能具有显著的增强作用。
This experiment was conducted to study the effects of Acanthopanax senticosus polysaccharide(ASPS) on distribution and immune function of T lymphocytes in spleen of immunosuppressed chicks,so as to evaluate the immune-enhancing effect of ASPS.A total of 300 4-day-age Hy-Line Brown male chicks were randomly divided into3 groups with 100 chicks in each group.Group 1 was the blank control group,group 2 was the cyclophosphamide(CTX)-induced immunosuppression group,and group 3 was the ASPS+CTX group.Groups 2 and 3 were subcutaneously injected CTX at a dose of 80 mg/kg·d for 3 consecutive days.From 10 days of age,group 3 was subcutaneously injected with 0.2 mL/bird of 200 mg/mL ASPS every day for 3 consecutive days,and group 1 and group 2 were injected with the same amount of sterilized normal saline in the same way.The spleens were taken at 7,14,21 and 28 days after injection,and the distribution and number of CD;a nd CD;T lymphocytes in the spleen were detected by immunohistochemical staining and flow cytometry,and the expression levels of IFN-γ and IL-2 mRNA in the spleen were detected by fluorescence quantitative PCR.The results showed that CTX significantly reduced the number of CD;and CD;T lymphocytes in the spleen(P<0.05),the number of PALS mainly composed of CD;cells decreased or disappeared,and the distribution range of CD;and CD;T lymphocytes in red and white pulp was reduced,and the cell arrangement was loose.mRNA expression levels of IFN-γ and IL-2 were significantly decreased compared with blank control group(P<0.05).Under the effect of ASPS,the number of CD;and CD;T lymphocytes in the ASPS+CTX group was significantly higher than that in the CTX group(P<0.05),the area of PALS was increased gradually with the increase of age,and the structure was gradually complete and clear.The distribution range of CD;T lymphocytes in ASPS+CTX group was significantly higher than that in CTX group,and the distribution of CD;cells in the red and white pulp was more intensive,and the level of CD;cells basically returned to normal leved at 28 days after immunization.mRNA expression levels of IFN-γ and IL-2 in ASPS+CTX group were significantly increased compared with CTX group(P<0.05).The results indicated that ASPS could significantly improve the abnormal location,distribution and number of CD;and CD;T lymphocytes in spleen caused by CTX,as well as the decreased expression levels of IFN-γ and IL-2 mRNA,which proved that ASPS had a significant enhancing effect on immune function of spleen in chickens.
作者
张英楠
刘洪章
杨树宝
ZHANG Yingnan;LIU Hongzhang;YANG Shubao(College of Life Science,Changchun Sci-Tech University,Changchun,Jilin 130600;Jilin Medical University,Jilin,Jilin 132013;College of Animal Science and Technology,Jilin Agricultural University,Changchun,Jilin 130118)
出处
《中国家禽》
北大核心
2021年第10期32-38,共7页
China Poultry
基金
吉林省教育厅“十三五”科学技术项目(JJKH20200355KJ、JJKH20201293JY)
国家自然科学基金资助项目(31372391)。
关键词
刺五加多糖
鸡
免疫抑制
T淋巴细胞
细胞因子
Acanthopanax senticosuspolysaccharide
chicken
immunosuppression
T lymphocytes
cytokine
作者简介
张英楠(1983-),女,博士,副教授,主要从事中药提取物免疫调节作用的研究,E-mail:zhangyingnan1983@163.com;通讯作者:杨树宝(1981-),男,博士,副教授,主要从事动物发育及免疫调节方面的研究,E-mail:yangshubao1981@163.com。
