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山羊ESC培养上清液对PBMC和PBL转化及分泌活性的影响
吴庆侠1, 杨普光1, 王爱华1
西北农林科技大学 动物医学院 农业部家畜生殖内分泌和胚胎工程重点开发实验室
摘要:
【目的】 探索山羊子宫内膜基质细胞(ESC)培养上清,对山羊外周血单核细胞(PBMC)和外周血淋巴细胞(PBL)的转化与分泌活性的影响。【方法】 分离山羊PBMC和PBL,制备原代ESC(ESC0)及永生化传至30代的ESC(ESC30)培养上清,将不同培养上清与PHA组合刺激PBMC与PBL转化,应用MTT法检测ESC0及ESC30培养上清对PBMC和PBL转化的作用,ELISA检测PBMC和PBL分泌IL-2、IL-4的相对含量。【结果】 ESC0及ESC30上清可刺激PBMC/PBL转化并分泌IL-2和IL-4,抑制PHA-P诱导的PBMC/PBL增殖及IL-2的分泌,但对PHA-P引起的PBMC/PBL分泌IL-4有促进或协同作用。【结论】 ESC能通过旁分泌作用影响PBMC和PBL的转化与分泌活性,优势活化Th2型淋巴细胞,在子宫局部免疫调节中发挥重要作用。
关键词:  山羊  原代子宫内膜基质细胞  传至30代的子宫内膜基质细胞  外周血单核细胞  外周血淋巴细胞  IL-2  IL-4
DOI:
分类号:
基金项目:国家自然科学基金项目(39770545); 西北农林科技大学青年骨干教师支持计划项目(01140305)
Effects of supernatant of goat endometrial stromal cells (ESC) on activation and secretion activity of PBMC and PBL
Abstract:
【Objective】 The study was to explore the effects of endometrial stroma cells (ESC) on proliferation and secretion activity of PBMC and PBL in goat via the way of paracrine. 【Method】 Peripheral blood mononuclear cell (PBMC)and peripheral blood leucocyte (PBL)were isolated,the supernatant of primary ESC (ESC0) and the 30th immortalized ESC(ESC30) were prepared.The effects of supernatant on proliferation of PBMC and PBL were detected by MTT respectively,and the effects on the secretion of IL-2 and IL-4 were determined by ELISA. 【Result】 The supernatant from both the ESC0 and the ESC30could stimulate the proliferation of PBMC and PBL and induce the secretion of IL-2 and IL-4,inhibit the proliferation and IL-2 secretion of PBMC/PBL induced by PHA-P,but enhance IL-4 secretion induced by PHA-P. 【Conclusion】 The results revealed that ESC affects the activation and secretion of PBMC and PBL,predominantly the Th2 lymphocytes,in a paracrine way,thus plays an important role in local uterine immune regulation.
Key words:  goat  ESC  ESC30,PBMC  PBL  IL-2  IL-4