摘要
探讨术芩总多糖对免疫低下小鼠脾淋巴细胞增殖与自噬的影响。采用CCK-8法筛选术芩总多糖对小鼠脾淋巴细胞增殖的最佳作用浓度及时间;试验分为空白组、3-MA(自噬抑制剂)组、3-MA+术芩总多糖组,Western blot测定术芩总多糖对小鼠脾淋巴细胞自噬相关蛋白Beclin1、LC3B表达的影响;试验分为空白组、环磷酰胺致免疫低下模型组、模型+术芩总多糖组,Western blot测定术芩总多糖对免疫低下小鼠脾淋巴细胞自噬相关蛋白Beclin1、LC3B表达的影响;试验结果:1)质量浓度为10^(-1)μg·mL^(-1)的术芩总多糖作用小鼠脾淋巴细胞12 h,细胞的增殖能力最强; 2)环磷酰胺免疫低下模型小鼠脾淋巴细胞增殖能力降低,术芩总多糖使淋巴细胞的增殖能力增强; 3)3-MA抑制小鼠脾淋巴细胞自噬时,自噬起始相关蛋白Beclin1的表达降低,术芩总多糖能使Beclin1的表达增加;当自噬相关蛋白LC3BⅡ/LC3BⅠ的表达升高时,术芩总多糖能降低LC3BⅡ/LC3BⅠ的表达; 4)免疫低下小鼠脾淋巴细胞自噬相关蛋白Beclin1、LC3BⅡ/LC3BⅠ的表达升高,术芩总多糖能降低Beclin1、LC3BⅡ/LC3BⅠ蛋白的表达。环磷酰胺降低小鼠免疫功能可能与其损伤脾淋巴细胞增殖发育,激活过度的自噬导致Ⅱ型程序性细胞死亡有关,术芩总多糖能抑制过高的自噬,增强淋巴细胞增殖能力,通过调控Beclin1、LC3BⅡ/LC3BⅠ蛋白的表达而调节机体的免疫功能。
To investigate the effect of polysaccharides from ZhuQin Formula(ZQp)on the proliferation and autophagy of spleen lymphocyte in immunocompromised mice,the optimum concentration and time of ZQp on the proliferation of spleen lymphocyte in mice was screened by CCK-8method.The test was divided into blank group,3-MA(autophagy inhibitor)group,3-MA+ZQp group,Western blot was used to determine the effect of ZQp on the expression of autophagy related protein Beclin1and LC3B in mice under the inhibition of3-MA;The test was divided into blank group,cyclophosphamide induced immunocompromised model group and model+ZQp group,Western blot was used to determine the effect of ZQp on the expression of autophagy related protein Beclin1and LC3B in immunocompromised mice.Test results were as follows:1)The ZQp with the concentration of10-1μg·mL-1acted on mouse spleen lymphocytes for12h,and the cell proliferation ability was the strongest;2)The proliferation ability of spleen lymphocyte in mice with cyclophosphamide immunodeficiency was reduced,and the proliferation of lymphocyte was enhanced by ZQp;3)When3-MA inhibited autophagy of spleen lymphocyte,the expression of autophagy related protein Beclin1decreased,and ZQp could increase the expression of Beclin1;When the expression of autophagy related protein LC3BⅡ/LC3BⅠincreased,ZQp could reduce the expression of LC3BⅡ/LC3BⅠ;4)The expression of autophagy related protein Beclin1and LC3BⅡ/LC3BⅠincreased in spleen lymphocytes of immunocompromised mice.ZQp could decrease the expression of Beclin1and LC3BⅡ/LC3BⅠprotein.The results suggest that:Cyclophosphamide-induced immune dysfunction in mice may be related to the splenic lymphocyte proliferation and development impair and excessive activation of autophagy lead to typeⅡprogrammed cell death.ZQp can inhibit excessive autophagy,enhance the proliferation of lymphocytes,and regulation the immune function by regulating the expression of Beclin1and LC3BⅡ/LC3BⅠprotein.
作者
庞敏
朱兆荣
乔芊芊
詹珠玉
刘娟
PANG Min;ZHU Zhao-rong;QIAO Qian-qian;ZHAN Zhu-yu;LIU Juan(College of Animal Science, Southwest University, Chongqing 402460, China;ChineseVeterinary Herbal Drugs Innovation Research Lab, University Veterinary Science EngineeringResearch Center in Chongqing, Chongqing 402460, China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2018年第12期2762-2770,共9页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家科技基础专项(2013FY110600-03)
公益性行业(农业)科研专项(201303040-05)
