| 杨 帆,吕 杰,王月秋,李 洋,王丽宏.LncRNA Kcnq1ot1/miR-214-3p/caspase-1通路调控高糖处理的心脏成纤维细胞焦亡[J].,2021,(10):1811-1817 |
| LncRNA Kcnq1ot1/miR-214-3p/caspase-1通路调控高糖处理的心脏成纤维细胞焦亡 |
| LncRNA Kcnq1ot1/miR-214-3p/caspase-1 Pathway Regulates Pyroptosis of High Glucose-Treated Cardiac Fibroblasts |
| 投稿时间：2021-01-06 修订日期：2021-01-28 |
| DOI：10.13241/j.cnki.pmb.2021.10.003 |
| 中文关键词: 长链非编码RNA Kcnq1ot1 高糖 焦亡 成纤维细胞 |
| 英文关键词: Long non-coding RNA Kcnq1ot1 High glucose Pyroptosis Cardiac fibroblasts |
| 基金项目:国家自然科学基金项目（81770809） |
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| 中文摘要: |
| 摘要 目的：探讨长链非编码RNA Kcnq1ot1在高糖处理的心脏成纤维细胞中调控焦亡的作用及具体机制。方法：培养C57BL/6乳鼠原代心脏成纤维细胞，分别用5.5 mM和30 mM葡萄糖培养，用免疫荧光、qRT-PCR和western blot方法检测NLRP3、caspase-1和IL-1β的表达。高糖处理的成纤维细胞抑制Kcnq1ot1，检测caspase-1的表达。生物信息学和荧光素酶报告基因检测验证与Kcnq1ot1和caspase-1存在共同互补结合位点的microRNA。应用qRT-PCR和western blot方法检测高糖诱导的心脏成纤维细胞干扰Kcnq1ot1后miR-214-3p的表达，以及过表达或干扰miR-214-3p后caspase-1的表达水平。高糖诱导的细胞单独干扰Kcnq1ot1或同时抑制Kcnq1ot1和miR-214-3p，检测caspase-1、NLRP3和IL-1?茁的表达水平。结果：高糖诱导的成纤维细胞中焦亡激活，Kcnq1ot1表达明显升高；抑制Kcnq1ot1后caspase-1表达显著下调。生物信息学和荧光素酶报告基因检测发现miR-214-3p与Kcnq1ot1和caspase-1存在共同互补结合位点。高糖诱导的心脏成纤维细胞干扰Kcnq1ot1后miR-214-3p表达升高；过表达 miR-214-3p 后caspase-1表达降低，抑制miR-214-3p后caspase-1表达升高。同时抑制Kcnq1ot1和miR-214-3p可逆转干扰Kcnq1ot1对caspase-1的降低作用。结论：干扰Kcnq1ot1能够通过抑制miR-214-3p/caspase-1信号转导通路，抑制高糖诱导的心脏成纤维细胞焦亡。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the role and mechanism of long non-coding RNA Kcnq1ot1 in regulating pyroptosis in high glucose-treated cardiac fibroblasts. Methods: Primary cardiac fibroblasts of C57BL/6 mice were divided into the control group (5.5 mM glucose) and the high glucose group (HG group, 30 mM glucose). The expression levels of NLRP3, caspase-1 and IL-1β were detected by immunofluorescence, qRT-PCR and western blot. Si-Kcnq1ot1 was transfected into high glucose-treated cardiac fibroblasts and the expression levels of caspase-1 were detected. Bioinformatics and luciferase assay were used to forecast the microRNAs that have the common complementary binding sites with both Kcnq1ot 1 and caspase-1. The expression of miR-214-3p after silencing Kcnq1ot1 and the expression of caspase-1 after transfection of miR-214-3p mimimics or AMO-214-3p in high glucose-treated cardiac fibroblasts were detected by qRT-PCR and western blot. The HG-treated fibroblasts were respectively transfected with si-NC+AMO-NC, si-Kcnq1ot1+AMO-NC and si-Kcnq1ot1+AMO-214-3p. The expression levels of caspase-1, NLRP3, IL-1β were detected. Results: Pyroptosis was activated and the expression of Kcnq1ot1 was elevated in high glucose-treated primary cardiac fibroblasts. Caspase-1 was downregulated after silencing Kcnq1ot1. Bioinformatics and luciferase assay reveal that miR-214-3p has common complementary binding sites with both Kcnq1ot1 and caspase-1. MiR-214-3p was increased after inhibiting Kcnq1ot1 in HG-treated cardiac fibroblasts, while caspase-1 was decreased after overexpression of miR-214-3p and was increased after inhibiting miR-214-3p. Reverse experiments showed that Kcnq1ot1 regulated caspase-1 through miR-214-3p. Conclusion: Silencing Kcnq1ot1 can alleviate high glucose-induced cardiac fibroblast pytoptosis by inhibiting miR-214-3p/caspase-1 signal pathway. |
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