Microvesicles Derived from Inflammation-Challenged Endothelial Cells Modulate Vascular Smooth Muscle Cell Functions.
Microvesicles Derived from Inflammation-Challenged Endothelial Cells Modulate Vascular Smooth Muscle Cell Functions.
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源自炎症挑战的内皮细胞的微泡调节血管平滑肌细胞功能
DOI：
10.3389/fphys.2016.00692
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发表时间：
2016
影响因子：
4
通讯作者：
Ma X
中科院分区：
文献类型：
--
作者：
Pan Q;Liu H;Zheng C;Zhao Y;Liao X;Wang Y;Chen Y;Zhao B;Lazartigues E;Yang Y;Ma X
目的：微泡（Microvesicles，MV）可通过转运其内含物调节受体细胞的功能.我们以前的研究表明，肿瘤坏死因子-α（TNF-α）和血清剥夺（SD）刺激的内皮祖细胞释放MV，对内皮细胞产生不利影响。在这项研究中，我们研究了内皮MV（EMV）对人脑血管平滑肌细胞（HBVSMC）增殖，迁移和凋亡的潜在影响。方法：体外培养人脑微血管内皮细胞（humanbrainmicrovascularendothelialcells，HBMEC），用TNF-α + SD培养液培养，制备EMV。通过用RNA酶A处理EMV以去除RNA来制备RNA酶-EMV。EMV和RNase-EMV分别与HBVSMC共培养后，检测HBVSMC增殖、凋亡和迁移能力。Mek 1/2抑制剂PD 0325901用于通路分析。Western blot检测Mek 1/2、Erk 1/2、磷酸化Erk 1/2、活化caspase-3和Bcl-2蛋白的表达。通过qRT-PCR测量miR-146 a-5 p的水平。结果：（1）EMV能显著促进HBVSMC增殖和迁移;（2）EMV可使HBVSMC凋亡率降低约35%，并伴有caspase-3的下调和Bcl-2的上调;（3）EMV可使HBVSMC中miR-146 a-5 p的表达增加约2倍;（4）RNase处理的EMV对HBVSMC活性和miR-146 a-5 p表达的影响低于EMV。结论：炎症刺激下产生的EMV可通过其携带的RNA调节HBVSMC的功能和命运。这与Mek 1/2/Erk 1/2通路的激活和caspase-3/Bcl-2的调节有关，在此过程中miR-146 a-5 p可能发挥重要作用。这些数据表明，EMV来源于炎症激发的内皮细胞是有害的HBVSMC的稳态功能，突出了血管疾病的潜在的新的治疗靶点。
Purpose: Microvesicles (MV) can modulate the function of recipient cells by transferring their contents. Our previous study highlighted that MV released from tumor necrosis factor-α (TNF-α) plus serum deprivation (SD)-stimulated endothelial progenitor cells, induce detrimental effects on endothelial cells. In this study, we investigated the potential effects of endothelial MV (EMV) on proliferation, migration, and apoptosis of human brain vascular smooth cells (HBVSMC). Methods: EMV were prepared from human brain microvascular endothelial cells (HBMEC) cultured in a TNF-α plus SD medium. RNase-EMV were made by treating EMV with RNase A for RNA depletion. The proliferation, apoptosis and migration abilities of HBVSMC were determined after co-culture with EMV or RNase-EMV. The Mek1/2 inhibitor, PD0325901, was used for pathway analysis. Western blot was used for analyzing the proteins of Mek1/2, Erk1/2, phosphorylation Erk1/2, activated caspase-3 and Bcl-2. The level of miR-146a-5p was measured by qRT-PCR. Results: (1) EMV significantly promoted the proliferation and migration of HBVSMC. The effects were accompanied by an increase in Mek1/2 and p-Erk1/2, which could be abolished by PD0325901; (2) EMV decreased the apoptotic rate of HBVSMC by approximately 35%, which was accompanied by cleaved caspase-3 down-regulation and Bcl-2 up-regulation; (3) EMV increased miR-146a-5p level in HBVSMC by about 2-folds; (4) RNase-treated EMV were less effective than EMV on HBVSMC activities and miR-146a-5p expression. Conclusion: EMV generated under inflammation challenge can modulate HBVSMC function and fate via their carried RNA. This is associated with activation of theMek1/2/Erk1/2 pathway and caspase-3/Bcl-2 regulation, during which miR-146a-5p may play an important role. The data suggest that EMV derived from inflammation-challenged endothelial cells are detrimental to HBVSMC homeostatic functions, highlighting potential novel therapeutic targets for vascular diseases.