miR-125b通过TLR4/NF-κB信号通路对心脏成纤维细胞增殖和迁移的影响

doi:10.13481/j.1671-587x.20200213

摘要/Abstract

摘要： 目的：探讨miR-125b通过Toll样受体4/核因子κB（TLR4/NF-κB）信号通路对心脏成纤维细胞增殖和迁移的影响，阐明miR-125b在心肌纤维化中的作用及机制。方法：将对数生长期HEH2细胞随机分为空白对照组、阴性对照组和miR-125b inhibitors组。miR-125b inhibitors组HEH2细胞转染miR-125b inhibitors，阴性对照组HEH2细胞转染阴性对照inhibitors，空白对照组HEH2细胞不进行转染。采用逆转录-聚合酶链反应（RT-PCR）法测定各组细胞中miR-125b水平，采用MTT法测定HEH2细胞增殖活性，Transwell小室测定HEH2细胞迁移能力，采用Western blotting法测定HEH2细胞中Ⅰ型胶原蛋白（Col Ⅰ）、Ⅲ型胶原蛋白（Col Ⅲ）、α平滑肌肌动蛋白（α-SMA）、TLR4和NF-κB蛋白表达水平。结果：与空白对照组和阴性对照组比较，miR-125b inhibitors组HEH2细胞中miR-125b水平降低（P<0.01），细胞增殖活性和迁移细胞数降低（P<0.05），Coll Ⅰ、Coll Ⅲ、α-SMA、TLR4和NF-κB蛋白表达水平降低（P<0.05）。空白对照组和阴性对照组HEH2细胞中miR-125b水平，细胞增殖活性，迁移细胞数，细胞中Coll Ⅰ、Coll Ⅲ、α-SMA、TLR4和NF-κB蛋白表达水平比较差异均无统计学意义（P>0.05）。结论：下调miR-125b水平可抑制心脏成纤维细胞增殖和迁移，其机制可能与TLR4/NF-κB信号通路有关。

关键词: miR-125b, Toll样受体4, 核因子κB, 成纤维细胞, 心肌纤维化

Abstract: Objective: To explore the effects of miR-125b on the proliferation and migration of cardiac fibroblasts via Toll-like receptor-4/nuclear factor-κB (TLR4/NF-κB) signaling pathway, and to clarify the role and mechanism of miR-125b in myocardial fibrosis. Methods: The HEH2 cells in logarithmic growth phase were randomly divided into blank control group, negative control group and miR-125b inhibitors group.The HEH2 cells in miR-125b inhibitors group were transfected with miR-125b inhibitors,and the HEH2 cells in negative control group were transfected with negative control inhibitors,and the HEH2 cells in blank control group were not transfected.The levels of miR-125b in the cells in various groups were determined by reverse transcription-polymerase chain reaction (RT-PCR) method.The proliferation activities of HEH2 cells were determined by MTTmethod.The Transwell chamber was used to measure the migration abilities of HEH2 cells;the expression levels of type Ⅰ collagen (Col Ⅰ), type Ⅲ collagen (Col Ⅲ), α-smooth muscle actin (α-SMA), TLR4 and NF-κB proteins in the HEH2 cells were determined by Western blotting method. Results: Compared with blank control group and negative control group, the level of miR-125b in the HEH2 cells in miR-125b inhibitors group was decreased (P<0.05), the proliferation activity and the number of migration cells were decreased (P<0.05), and the expression levels of Col Ⅰ, Col Ⅲ, α-SMA, TLR4, and NF-κB proteins were decreased (P<0.05).There were no significant differences in the miR-125b levels,the proliferation activities,the number of migration cells,the expression levels of Coll Ⅰ, Coll Ⅲ, α-SMA, TLR4, and NF-κB proteins in the HEH2 cells between blank control group and negative control group(P>0.05). Conclusion: Down-regulation of miR-125b level can inhibit the proliferation and migration of the cardiac fibroblasts, and its mechanism may be related to the TLR4/NF-κB signaling pathway.

Key words: miR-125b, Toll-like receptor 4, nuclear factor-κB, fibroblasts, myocardial fibrosis

中图分类号:

R329.2

万琦, 余宝刚. miR-125b通过TLR4/NF-κB信号通路对心脏成纤维细胞增殖和迁移的影响[J]. 吉林大学学报(医学版), 2020, 46(02): 286-291.

WAN Qi, YU Baogang. Effects of miR-125b on proliferation and migration of cardiac fibroblasts by TLR4/NF-κB signaling pathway[J]. Journal of Jilin University(Medicine Edition), 2020, 46(02): 286-291.

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