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目的:探讨脂多糖(LPS)及IL-13对系膜细胞表达IL-12的影响作用。方法:用酶联免疫吸附(ELISA)法检测LPS对系膜细胞分泌IL-12的影响,实验分组:①LPS(20μg/ml)对系膜细胞刺激不同时间;②不同剂量LPS刺激系膜细胞24小时。用ELISA法和逆转录聚合酶链反应(RT-PCR)法分别检测IL-13对系膜细胞IL-12蛋白和IL-12p40 mRNA表达的影响,分空白对照组,LPS(10μg/ml)刺激组及实验组(LPS10μg/ml+不同浓度的IL-13)。结果:未受LPS刺激的系膜细胞几乎不分泌IL-12,在一定的剂量范围内,LPS诱导系膜细胞显著表达IL-12,但随着LPS刺激时间及浓度的增加,IL-12的表达量反而减少。IL-13在1~100ng/ml浓度范围内呈剂量依赖性抑制LPS诱导的系膜细胞IL-12分泌及其mRNA表达(P<0·01)。结论:LPS可诱导系膜细胞表达IL-12,但系膜细胞在表达IL-12时对LPS的长期及高浓度刺激产生耐受性。IL-13可能通过抑制LPS诱导系膜细胞分泌IL-12,作为抗炎性细胞因子在肾小球肾炎发病机制中发挥一定作用。
Objective: To investigate the effect of lipopolysaccharide (LPS) and IL-13 on the expression of IL-12 in mesangial cells. Methods: The effect of LPS on the secretion of IL-12 by mesangial cells was detected by enzyme-linked immunosorbent assay (ELISA). The experimental groups were as follows: ① LPS (20μg / ml) stimulated mesangial cells for different time; ② LPS stimulated mesangial cells 24 hour. The effects of IL-13 on the expression of IL-12 and IL-12p40 mRNA in mesangial cells were detected by ELISA and reverse transcription-polymerase chain reaction (RT-PCR) Group and experimental group (LPS10μg / ml + different concentrations of IL-13). Results: LPS-stimulated mesangial cells seldom secrete IL-12. LPS induced significant IL-12 expression in a dose-dependent manner in LPS-stimulated mesangial cells. However, with the increase of LPS stimulation time and concentration, Instead, the amount of expression decreases. IL-13 inhibited IL-12 secretion and its mRNA expression in LPS-induced mesangial cells in a concentration range of 1 ~ 100ng / ml (P <0.01). CONCLUSIONS: LPS induces the expression of IL-12 in mesangial cells, but the mesangial cells are resistant to long-term and high-concentration LPS stimulation when IL-12 is expressed. IL-13 may play an important role in the pathogenesis of glomerulonephritis by inhibiting the secretion of IL-12 by mesangial cells induced by LPS, as an anti-inflammatory cytokine.