中国寄生虫学与寄生虫病杂志

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蛔虫粗抗原对人肺腺癌细胞凋亡中白细胞介素6和转化生长因子β分泌的影响

黄川1,2,黄艳琴2,杨志1,2,胡银英2,戴志芳2,袁芳2,袁铿2 *   

  1. 1 南昌大学医学院,南昌 330006;2 江西省医学科学研究院,南昌 330006
  • 出版日期:2015-06-30 发布日期:2015-09-08

Effect of Crude Antigens from Ascaris lumbricoides on the Apoptosis and Secretion of IL-6 and TGF-β of Human Lung Adenocarcinoma Cells

HUANG Chuan 1,2, HUANG Yan-qin 2, YANG Zhi 1, 2, HU Yin-ying 2,  DAI Zhi-fang 2, YUAN Fang 2, YUAN Keng 2 *   

  1. 1  Medical College, Nanchang University, Nanchang 330006, China; 2 Jiangxi Academy of Medical Sciences, Nanchang 330006, China
  • Online:2015-06-30 Published:2015-09-08

摘要:

【摘要】  目的  探讨蛔虫粗抗原对人肺腺癌细胞株(A549)凋亡,以及白细胞介素(IL-6)和转化生长因子β(TGF-β)分泌的影响。  方法  收集新鲜蛔虫虫体,制备蛔虫粗抗原。将A549细胞分别与低、中、高浓度的蛔虫粗抗原(蛋白浓度分别为25、125和500 μg/ml)共培养,即低浓度组、中浓度组和高浓度组,同时设A549细胞+培养液的阴性对照组和阿霉素阳性对照组。用流式细胞术、ELISA和实时定量PCR(qPCR)分别检测作用1、18和24 h后A549细胞凋亡率和细胞周期,以及IL-6和TGF-β细胞因子水平和mRNA水平。  结果  3个浓度的蛔虫粗抗原作用于细胞1、18和24 h后,细胞凋亡率均显著高于同时间点的阴性对照组(P<0.01)。其中,中浓度组作用18 h后细胞凋亡率最高,为(47.10±3.68)%。细胞周期检测结果显示,中浓度组作用18 h后,G0/G1期细胞比例升高,S期细胞比例下降,与阴性对照组间差异有统计学意义(P<0.01)。ELISA检测结果显示,相同作用时间下,A549细胞IL-6水平大多随蛔虫粗抗原作用浓度的增加而升高,TGF-β水平随着蛔虫粗抗原浓度和作用时间的不同,无明显的变化趋势。qPCR结果显示,中浓度组A549细胞的IL-6和TGF-β mRNA水平在作用18 h后达到最高,分别为5.95±0.31和3.43±0.35,且显著高于阴性对照组(P<0.01)。  结论  蛔虫粗抗原可诱导A549细胞凋亡,并将细胞阻滞在G0/G1期,其调节过程可能有IL-6和TGF-β的参与。

关键词:  人蛔虫, 粗抗原, 白细胞介素6, 转化生长因子&beta, A549细胞, 细胞凋亡

Abstract:

【Abstract】 Objective  To investigate the effect of crude antigens of Ascaris lumbricoides on the secretion of IL-6 and TGF-β of human lung adenocarcinoma cells (A549 cells), and the apoptosis of A549 cells.  Methods  Crude antigens of A. lumbricoides were prepared. A549 cells were co-cultured with 25, 125, and 500 μg/ml crude antigens of A. lumbricoides for 1, 18, and 24 h, named as low concentration group, medium concentration group, and high concentration group, respectively. Meanwhile, A549 cells were co-cultured with culture medium (negative control) and 12.5 μg/ml adriamycin (positive control). The apoptosis rate was detected by using Annexin V-FITC apoptosis detection kit. The cell changes were determined by flow cytometry. The levels of mRNA expression of IL-6 and TGF-β were detected by ELISA and real time PCR, respectively.  Results  The apoptosis rate of A549 cells induced by crude antigens for 1, 18, and 24 h was significantly higher than that of negative control (P<0.01). The apoptosis rate in medium concentration group (treated for 18 h) was highest [(47.10±3.68)%]. After co-culture with 125 μg/ml crude antigens for 18 h, the proportion of G0/G1 phase cells increased and that of S phase cells decreased, and there was a significant difference between medium concentration group and negative control group. At the same time, the level of IL-6 increased with the increasing concentration of crude antigens. However, no significant difference was found in the level of TGF-β among the groups. In the medium concentration group, mRNA expression levels of IL-6 (5.95±0.31) and TGF-β(3.43±0.35) of A549 cells reached peak on the 18th hour, and were significantly higher than that of the control (P<0.01).  Conclusion  The cell cycle of A549 cells is blocked in G0/G1 phase induced by crude antigens of A. lum-bricoides. And the apoptosis may be related to the changes in the level of TGF-β and IL-6.

Key words: Ascaris lumbricoides, Crude antigen, IL-6, TGF-β, A549 cell, Apoptosis