Proliferation of Vascular Smooth Muscle Cells under ox-LDL Is Regulated by Alismatis rhizoma Decoction via InhibitingERK1/2 and miR-17∼92a Cluster Activation
Proliferation of Vascular Smooth Muscle Cells under ox-LDL Is Regulated by Alismatis rhizoma Decoction via InhibitingERK1/2 and miR-17∼92a Cluster Activation
复制标题
泽泻汤通过抑制ERK1/2和miR-17-92a簇激活调控ox-LDL作用下血管平滑肌细胞的增殖
DOI：
10.1155/2020/7275246
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发表时间：
2020-08-24
影响因子：
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通讯作者：
Xue, Xiehua
中科院分区：
文献类型：
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作者：
Shen, Julian;Wei, Wei;Xue, Xiehua
背景：泽泻根茎汤（AD）具有抗动脉粥样硬化活性。AD对血管平滑肌细胞（VSMC）增殖的抑制作用尚不清楚。目标。探讨AD对氧化低密度脂蛋白（ox-LDL）诱导的VSMC增殖的影响及其机制。材料和方法。雄性SD大鼠分别口服AD （2.56 g/mL）或0.9% NaCl灌胃4 mL，每日2次，连续7 d。然后采集含ad血清（adc）。采用MTS法测定VSMC活力。5-溴脱氧尿苷（BrdU）免疫细胞化学检测VSMCs的增殖。进行microRNA （miRNA）分析，并从TargetScan 7.2数据库中搜索miRNA的靶基因。采用western blotting或定量反转录PCR检测基质金属蛋白酶-2/9 （MMP-2/9）、细胞周期蛋白D1/E、细胞周期蛋白依赖性激酶抑制剂1B （p27）、细胞外调节蛋白激酶1/2 （ERK1/2）和ERK1/2磷酸化的表达。结果。ox- ldl诱导的miR-17-92a表达促进VSMC增殖。AD和ERK1/2抑制剂U0126 （10 μmol/L）抑制VSMC增殖，降低miR-17 ~ 92a的过表达。AD抑制了VSMCs中ERK1/2的磷酸化，降低了MMP-2/9的表达。AD联合ERK1/2抑制剂抑制细胞周期蛋白D1/E的表达，p27升高。根据TargetScan 7.2数据库，miR-17 ~ 92a的靶基因作用于参与动脉粥样硬化过程的金属蛋白酶(TIMPs)-MMPs、p27/21细胞周期蛋白和过氧化物酶体增殖因子激活受体α (PPARα) atp结合盒转运体（ABC） A1/G1的组织抑制剂。结论。AD通过抑制ERK1/2和miR-17 ~ 92a活化抑制ox- ldl诱导的VSMC增殖。这些结果为AD治疗动脉粥样硬化提供了多靶点机制。对心脑血管疾病的治疗有一定的帮助。
Context: Alismatis rhizome decoction (AD) exhibits antiatherosclerotic activities. The activity of AD against vascular smooth muscle cell (VSMC) proliferation remains unclear. Objective. The mechanisms and effects of AD on oxidized low-density lipoprotein (ox-LDL)-induced VSMC proliferation were explored. Materials and methods. The male SD rats were fed with AD (2.56 g/mL) or 0.9% NaCl by oral gavage 4 mL twice daily for 7 d. Then, AD-containing serum (ADcs) was collected. MTS assay was applied to measure the VSMC viability. The proliferation of VSMCs was detected by 5-bromodeoxyuridine (BrdU) immunocytochemistry. The microRNA (miRNA) profiling was performed, and the target genes of miRNAs were searched from the TargetScan 7.2 database. The expressions of matrix metalloproteinases-2/9 (MMP-2/9), cyclin D1/E, cyclin-dependent kinase inhibitor 1B (p27), extracellular regulated protein kinases 1/2 (ERK1/2), and ERK1/2 phosphorylation were examined by western blotting or quantitative reverse transcription PCR. Results. The ox-LDL-induced miR-17-92a expression promoted VSMC proliferation. AD and the ERK1/2 inhibitor U0126 (10 μmol/L) inhibited VSMC proliferation and reduced the overexpression of miR-17∼92a. AD was found to inhibit phosphorylation of ERK1/2 and reduced the expression of MMP-2/9 in VSMCs. The expression of cyclin D1/E was suppressed, and p27 was elevated following treatment with AD as well as ERK1/2 inhibitor. According to the TargetScan 7.2 database, the target genes of miR-17∼92a act on tissue inhibitors of metalloproteinases (TIMPs)-MMPs, p27/21 cyclins, and peroxisome-proliferator-activated receptor α (PPARα) ATP-binding cassette transporter (ABC) A1/G1, which are involved in the process of atherosclerosis. Conclusions. AD inhibits ox-LDL-induced VSMC proliferation via inhibiting ERK1/2 and miR-17∼92a activation. The results provide the multitarget mechanisms for application of AD in the treatment of atherosclerosis. It would be helpful to the treatment of cardiovascular and cerebral diseases.