MicroRNA-199a-5p Regulates the Proliferation of Pulmonary Microvascular Endothelial Cells in Hepatopulmonary Syndrome
MicroRNA-199a-5p Regulates the Proliferation of Pulmonary Microvascular Endothelial Cells in Hepatopulmonary Syndrome
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MicroRNA-199a-5p 调节肝肺综合征肺微血管内皮细胞的增殖
DOI：
10.1159/000430252
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发表时间：
2015-01-01
影响因子：
--
通讯作者：
Yi, Bin
中科院分区：
文献类型：
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作者：
Zeng, Jing;Chen, Lin;Yi, Bin
背景/目标：肺微血管内皮细胞（PMVEC）增殖和血管生成参与了肝病综合征（HPS）的发生发展。microRNA-199 a-5 p（miR-199 a-5 p）是一种有效的血管生成调节因子，在肝病患者血清中的表达水平显著降低。然而，miR-199 a-5 p是否能调控PMVEC增殖尚未见报道。在此，我们描述了HPS中控制PMVEC增殖的miR-199 a-5 p。方法：用胆总管结扎（CBDL）大鼠血清或假手术大鼠血清处理PMVEC。使用miR-199 a-5 p模拟物或抑制剂来改变miR-199 a-5 p表达。使用SiRNA进行小窝蛋白-1（Cav-1）的敲除。NSC-23766用于抑制Rac 1活性。通过qRT-PCR和蛋白质印迹定量基因和蛋白质表达。3 H-TdR掺入法和CCK-8法检测细胞增殖。免疫荧光法检测应力纤维。结果：CBDL大鼠血清可诱导miR-199 a-5 p表达下调。miR-199 a-5 p的递送抑制CBDL大鼠血清诱导的PMVEC增殖，而miR-199 a-5 p的敲低促进PMVEC增殖。这分别伴随着Cav-1表达的减少和增加。Cav-1 siRNA可抑制miR-199 a-5 p对PMVEC增殖的促进作用。尽管在Cav-1缺陷细胞中应力纤维被破坏，但NSC-23766增加了应力纤维并有助于细胞增殖。结论：CBDL大鼠血清可诱导PMVECs中miR-199 a-5 p表达下调，导致Cav-1基因表达增加。Cav-1表达增加，通过抑制Rac 1活性，导致应力纤维的形成，这有助于PMVEC增殖，从而导致HPS的发病机制。
Background/Aims: Pulmonary microvascular endothelial cell (PMVEC) proliferation and angiogenesis contribute to the development of hepatopulmonary syndrome (HPS). MicroRNA-199a-5p (miR-199a-5p) has emerged as a potent regulator of angiogenesis, and its expression levels significantly decrease in the serum of patients with hepatopathy. However, it has not been reported about whether miR-199a-5p might control PMVEC proliferation. Here, we described the miR-199a-5p governing PMVEC proliferation in HPS. Methods: PMVECs were treated with rat serum from common bile duct ligation (CBDL) or sham. MiR-199a-5p mimic or inhibitor was used to change the miR-199a-5p expression. Knockdown of caveolin-1 (Cav-1) was performed using siRNA. NSC-23766 was used to inhibit Rac1 activity. Gene and protein expressions were quantified by qRT-PCR and western blot. Cell proliferation was analyzed by 3H-TdR incorporation and CCK-8 assays. Stress fibers were detected by immunofluorescence. Results: CBDL rat serum induced the down-regulation of miR-199a-5p. Delivery of miR-199a-5p suppressed the CBDL rat serum-induced PMVEC proliferation whereas knockdown of miR-199a-5p promoted PMVEC proliferation. This was accompanied by a decrease and an increase in Cav-1 expression, respectively. Cav-1 siRNA abolished the enhancement of PMVEC proliferation induced by the miR-199a-5p inhibition. Although stress fibers were disrupted in Cav-1 deficient cells, NSC-23766 increased stress fibers and contributed to cell proliferation. Conclusions: CBDL rat serum induced down-regulation of miR-199a-5p in PMVECs, which led to an increase of Cav-1 gene expression. Increased Cav-1 expression, by inhibiting Rac1 activity, led to the formation of stress fibers, which contribute to PMVEC proliferation and thus the pathogenesis of HPS.