体外研究人脐带血间充质干细胞对大鼠枯否细胞极化的影响

发布时间：2018-03-22 01:17

本文选题：枯否细胞　切入点：人间充质干细胞　出处：《安徽医科大学学报》2016年01期 　论文类型：期刊论文

【摘要】：目的采用脂多糖(LPS)诱导大鼠枯否细胞(KCs)发生极化改变,之后用人脐带血间充质干细胞(huM SCs)与LPS诱导的KCs在Transwell内共培养,以观察huM SCs对KCs极化偏移的调节作用。方法实验分为KCs组、KCs+LPS组、KCs+LPS+MSCs组。对各组的白介素-4(IL-4)、肿瘤坏死因子(TNF-α)、白介素-10(IL-10)、白介素-6(IL-6)等上清因子采用ELISA法进行检测;诱导型一氧化氮合酶(iNOS)、精氨酸酶-1(Arg-1)、信号传导与转录激活因子3、6(STAT-3、STAT-6)、核因子kappaB(NF-κB)用Western bolt进行检测,同时用荧光实时定量PCR(qRT-PCR)对以上结果进行验证。结果 KCs+LPS组促炎因子TNF-α、IL-6分泌增加,KCs+LPS+MSCs组抑炎因子IL-10、IL-4分泌增加;而Western blot检测表明,KCs+LPS组中iN OS升高,NF-κB p65入核增高;而KCs+LPS+MSCs组高表达Arg-1,同时pS TAT-3、pS TAT-6表达增加。结论 huM SCs能诱导已经发生M1极化的KCs向M2表型偏移,考虑可能与huM SCs分泌细胞因子有关,起到一种免疫调节作用,huM SCs调节巨噬细胞极化的分子机制可能与通过JAK-STAT信号转导通路有关。
[Abstract]:Objective to induce the polarization change of Kupffer cells in Kupffer cells induced by lipopolysaccharide (LPS), and then co-culture KCs induced by LPS and human umbilical cord blood mesenchymal stem cells (HUMSCs) in Transwell. Methods the effects of huM SCs on KCs polarization shift were observed. Methods the supernatant factors, such as IL-4, TNF- 伪, IL-10, IL-6 and IL-6, were measured by ELISA method in KCs group (KCs LPS group). Inducible nitric oxide synthase (iNOS), argininase 1 (Arg-1), signal transduction and transcriptional activator (3) STAT-3 (STAT-6), and nuclear factor kappa B (NF- 魏 B) were detected by Western bolt. Results the secretion of proinflammatory factor TNF- 伪 and IL-6 in KCs LPS group was increased, while IL-10 and IL-4 secretion were increased in KCs LPS MSCs group, while Western blot test showed that iNOS increased NF- 魏 B p65 in KCs LPS group, which was also verified by real-time fluorescence quantitative PCRQRT-PCR.Results in KCs LPS group, the secretion of proinflammatory factor TNF- 伪 and IL-6 increased, while in KCs LPS MSCs group, the secretion of IL-10 and IL-4 was increased, while that in Western blot group was increased. In KCs LPS MSCs group, the expression of Arg-1 was highly expressed, and the expression of pS TAT-3pS TAT-6 was increased. Conclusion huM SCs can induce KCs with M1 polarization to M2 phenotypic shift, which may be related to the secretion of cytokines by huM SCs. The molecular mechanism of hum SCs regulating macrophage polarization may be related to JAK-STAT signal transduction pathway.
【作者单位】：安徽医科大学第三附属医院(合肥市第一人民医院)消化内科;安徽医科大学公共卫生学院;
【基金】：国家自然科学基金(编号:81101272)
【分类号】：R575

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