川芎嗪通过上调人脐静脉内皮细胞中 miR-135b 促进缺氧处理后的细胞生长。
Ligustrazine promoted hypoxia-treated cell growth by upregulation of miR-135b in human umbilical vein endothelial cells.
机构信息
Operating Room, Jining No.1 People's Hospital, Jining 272011, China; Affiliated Jining No.1 People's Hospital of Jining Medical University, Jining Medical University, Jining 272067, China.
Operating Room, Jining No.1 People's Hospital, Jining 272011, China; Affiliated Jining No.1 People's Hospital of Jining Medical University, Jining Medical University, Jining 272067, China.
出版信息
Exp Mol Pathol. 2019 Feb;106:102-108. doi: 10.1016/j.yexmp.2018.12.005. Epub 2018 Dec 18.
BACKGROUND
Pressure ulcers are a kind of troublesome disease which caused by long-term pressure and subsequently lead to tissue festering necrosis because of sustained ischemia, hypoxia and malnutrition. In our study, we used hypoxia to stimulate human umbilical vein endothelial cells (HUVECs) to mimic pressure ulcers and investigated the effects of Ligustrazine (Lig) with multi-activities on HUVECs.
METHODS
HUVECs were treated by hypoxia to induce cell injury. HUVECs were administrated with Lig and/or transfected with miR-135b inhibitor or negative control. Cell viability and cell apoptosis were detected by Cell Counting kit-8 assay and flow cytometry, respectively. The protein expression of Cyclin D1 and p53, the apoptosis-related proteins (Bcl-2, Bax, pro-/Cleaved-Caspas-3), and the JNK/SAPK and PI3K/AKT/mTOR pathways related proteins was examined by western blot.
RESULTS
Hypoxia-induced injury presented by decreasing cell viability and increasing cell apoptosis. Then Lig administration enhanced cell viability and inhibited cell apoptosis. Importantly, miR-135b was upregulated by the treatment of Lig. Further studies revealed that transfection with miR-135b inhibitor led to the opposite result with decreasing cell viability and increasing cell apoptosis. In addition, Lig increased the phosphorylation of JNK, SAPK, PI3K, AKT and mTOR.
CONCLUSION
Lig promoted hypoxia-treated HUVECs cell growth as evidenced by increasing cell viability and reducing cell apoptosis. This process might be modulated by upregulation of miR-135b and subsequent activation of JNK/SAPK and PI3K/AKT/mTOR pathways.
背景
压疮是一种由长期压力引起的麻烦疾病，由于持续缺血、缺氧和营养不良，导致组织溃烂坏死。在我们的研究中，我们使用缺氧来刺激人脐静脉内皮细胞（HUVEC）模拟压疮，并研究了具有多种活性的川芎嗪（Lig）对 HUVEC 的影响。
方法
用缺氧处理 HUVEC 以诱导细胞损伤。用 Lig 和/或 miR-135b 抑制剂或阴性对照转染 HUVEC。通过细胞计数试剂盒-8 测定和流式细胞术分别检测细胞活力和细胞凋亡。通过 Western blot 检测细胞周期蛋白 D1 和 p53 的蛋白表达、凋亡相关蛋白（Bcl-2、Bax、pro-/Cleaved-Caspase-3）以及 JNK/SAPK 和 PI3K/AKT/mTOR 通路相关蛋白的表达。
结果
缺氧诱导的损伤表现为细胞活力降低和细胞凋亡增加。然后 Lig 给药增强了细胞活力并抑制了细胞凋亡。重要的是，Lig 处理后 miR-135b 上调。进一步的研究表明，转染 miR-135b 抑制剂导致相反的结果，即细胞活力降低和细胞凋亡增加。此外，Lig 增加了 JNK、SAPK、PI3K、AKT 和 mTOR 的磷酸化。
结论
Lig 通过增加细胞活力和减少细胞凋亡来促进缺氧处理的 HUVEC 细胞生长。该过程可能通过上调 miR-135b 并随后激活 JNK/SAPK 和 PI3K/AKT/mTOR 通路来调节。
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