枸橼酸对内毒素刺激下单核细胞细胞因子分泌的影响

doi:10.3969/cndt.j.issn.1006-298X.2018.01.008

肾脏病与透析肾移植杂志 ›› 2018, Vol. 27 ›› Issue (1): 41-45.DOI: 10.3969/cndt.j.issn.1006-298X.2018.01.008

枸橼酸对内毒素刺激下单核细胞细胞因子分泌的影响

出版日期:2018-02-28 发布日期:2018-03-01

Effect of citrate on lipopolysaccharideinduced monocyte cytokines release

Online:2018-02-28 Published:2018-03-01

摘要/Abstract

摘要：

目的：通过体外细胞培养，观察不同浓度枸橼酸对内毒素(LPS)诱导的单核细胞分泌细胞因子功能的影响。
方法：采用标准方法体外培养人类单核细胞株(THP1)，加入适量枸橼酸三钠(TSC)使其体外培养液TSC浓度分别为0、05 mmol/L、10 mmol/L，并给予不同浓度LPS(005 μg/ml、01 μg/ml、1 μg/ml)刺激，分别于刺激后6h、12h、24h、48h用CCK8试剂盒测定细胞活性、ELISA试剂盒测定上清液细胞因子白细胞介素6(IL6)、肿瘤坏死因子α(TNFα)水平及血气分析仪测定培养基内离子钙(Ca2+)水平。使用正常培养条件下的单核细胞做为对照组。
结果：不同浓度TSC对单核细胞活性无抑制作用(P>005)，但对不同时间点LPS诱导的单核细胞分泌细胞因子(TNFα和IL6)功能有一定影响，培养基TSC浓度05 mmol/L和10 mmol/L时，培养12h和24h TNFα的分泌量显著高于对照组(P<001)；TSC浓度为05 mmol/L时，IL6在24h开始出现分泌增高趋势，48h达到峰值(P<005)，TSC浓度10 mmol/L显著抑制IL6分泌(P<001)。
结论：本实验采用的TSC浓度不存在细胞毒性，单核细胞在此浓度下，受LPS刺激时表现出TNFα分泌亢进的现象，而IL6的分泌则表现出TSC低浓度下亢进，高浓度下受抑制的趋势。

关键词: 枸橼酸浓度, 单核细胞, 细胞因子

Abstract:

Ｏｂｊｅｃｔｉｖｅ：To investigate the effects of various concentrations of citrate on cytokines release by monocyte under stimulation of lipopolysaccharide(LPS) in vitro.
Ｍｅｔｈｏｄｏｌｏｇｙ：Human acute monocyte leukemia cell line(THP1) cells were cultured in vitro with different concentrations of trisodium citrate (TSC,0,05,10 mmol/L),and were stimulated with different concentrations of LPS(0,005,01,1 μg/ml).Cell samples and supernatants were collected 6h,12h,24h and 48h after cell culture.The monocyte viability,concentrations of interleukine6(IL6),tumor necrosis factorα (TNFα) and ionized calcium(Ca2+) were observed.
Ｒｅｓｕｌｔｓ：Various concentrations of citrate did not inhibit the viability of monocyte (P>005).The monocyte TNFα production after LPS stimulation was augmented with 05 mmol/L and 1 mmol/l citrate at 12h and 24h when compared with no citrate(P<001).The production of IL6 was augmented with 05 mmol/L citrate at 24h and 48h(P<005)，but was inhibited with 10 mmol/L citrate when compared with no citrate (P<001).
Ｃｏｎｃｌｕｓｉｏｎ：
Citrate in the investigated concentrations has no cytotoxicity to monocyte.After stimulation by LPS,production of TNFα was augmented with citrate,while production of IL6 was augmented only with low concentration of citrate but was inhibited with high concentration of citrate.

Key words: citrate concentration, monocyte, cytokine

周蕾，卢颖辉，徐斌，等. 枸橼酸对内毒素刺激下单核细胞细胞因子分泌的影响[J]. 肾脏病与透析肾移植杂志, 2018, 27(1): 41-45.

ZHOU Lei，LU Yinghui，XU Bing，et al. Effect of citrate on lipopolysaccharideinduced monocyte cytokines release[J]. Chinese Journal of Nephrology, Dialysis & Transplantation, 2018, 27(1): 41-45.

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ISSN 1006-298X CN 32-1425/R

枸橼酸对内毒素刺激下单核细胞细胞因子分泌的影响

Effect of citrate on lipopolysaccharideinduced monocyte cytokines release

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