[牛磺酸通过促进线粒体自噬抑制巨噬细胞M2极化]。

Chengying Chen, Chunhua Lan, Jianglang Yuan, Xingxing Kong, Li Lan, Xinhang Wang, Shengboxiaoji Chang, Cailing Lu, Xiyi Li, Shen Tang
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引用次数: 0

摘要

目的探讨牛磺酸调节M2巨噬细胞自噬极化的分子机制。方法将THP-1细胞分为4组:M0组(THP-1细胞经100 nmol/L肉豆酸酯作用48 h后极化为M0)、M2组(20 ng/mL干扰素-4 (IL-4)作用48 h诱导THP-1细胞极化为M2巨噬细胞)、M2联合牛磺酸组(在M2巨噬细胞基础上分别添加40或80 mmol/L牛磺酸)。采用实时荧光定量PCR检测M2巨噬细胞中甘露糖受体C-1 (MRC-1)、C-C基序趋化因子配体22(CCL22)和树突状细胞特异性ICAM-3捕获非整合素(CD209)的mRNA表达。采用线粒体和溶酶体探针,通过多功能酶标仪和共聚焦激光扫描显微镜检测线粒体和溶酶体的数量。采用JC-1型线粒体膜电位测定试剂盒检测线粒体膜电位(MMP)水平。Western blot检测线粒体自噬相关蛋白pten诱导的推定激酶1 (PINK1)和微管相关蛋白1轻链3 (LC3)的表达。结果与M0组比较,M2组细胞中MRC-1、CCL22、CD209、PINK1的表达、线粒体数量和MMP水平均显著升高,溶酶体数量和LC3II/LC3I比值均显著降低。与M2组比较,M2联合牛磺酸组MRC-1、CCL22、CD209的表达量、线粒体数量和MMP水平均显著下降,溶酶体数量增加,PINK1蛋白表达量和LC3II/LC3I比值升高。结论牛磺酸可通过降低M2巨噬细胞MMP水平、提高线粒体自噬水平、减少线粒体数量、抑制极化标志物mRNA表达等途径调控M2巨噬细胞的极化,防止过度极化。
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[Taurine inhibits M2 polarization of macrophages by promoting mitophagy].

Objective To investigate the molecular mechanism of taurine regulating the polarization of M2 macrophages by mitophagy. Methods THP-1 cells were divided into four groups: M0 group (THP-1 cells were treated by 100 nmol/L phorbol myristate ester for 48 hours to polarize into M0), M2 group (THP-1 cells were induced to polarize into M2 macrophages by 20 ng/mL interferon-4 (IL-4) for 48 hours), M2 combined with taurine groups (added with 40 or 80 mmol/L taurine on the basis of M2 macrophages). The mRNA expression of mannose receptor C type 1(MRC-1), C-C motif chemokine ligand 22(CCL22) and dendritic cell-specific ICAM-3 grabbing non-integrin (CD209) in M2 macrophages were detected by quantitative real-time PCR. Mitochondrial and lysosome probes were used to detect the number of mitochondria and lysosomes by multifunction microplate reader and confocal laser scanning microscope. The level of mitochondrial membrane potential (MMP) was detected by JC-1 MMP assay kit. The expression of mitophagy-related proteins PTEN-induced putative kinase 1 (PINK1) and microtubule-associated protein 1 light chain 3 (LC3) were detected by Western blot analysis. Results Compared with M0 group, the expression of MRC-1, CCL22, CD209 and PINK1, the number of mitochondria and the level of MMP in M2 group were significantly increased, whereas the number of lysosomes and LC3II/LC3I ratio were decreased. Compared with M2 group, the expressions of MRC-1, CCL22 and CD209, the number of mitochondria and the level of MMP in M2 combined with taurine group dropped significantly while the number of lysosomes was found increased, and the protein expression of PINK1 and LC3II/LC3I ratio were also increased. Conclusions The polarization of M2 macrophages is regulated by taurine to prevent excessive polarization via reducing the level of MMP, improving the level of mitophagy, reducing the number of mitochondria, and inhibiting the mRNA expression of polarization markers in M2 macrophages.

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