miR-30b 在高磷诱导的血管平滑肌细胞凋亡中的作用
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河北医科大学第四医院 肾内科,石家庄 050011

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R-33

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High phosphorus promotes apoptosis of vascular smooth muscle cells by inhibiting miR-30b
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Department of Nephrology, the Forth Hospital of Hebei Medical University, Shijiazhuang 050011, China

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    摘要:

    目的 观察微小 RNA30b(microRNA 30b,miR-30b)在高磷诱导的血管平滑肌细胞( vascular smooth muscle cells,VSMCs)凋亡中的作用及机制。 方法 体外培养大鼠胸主动脉 VSMCs,分为正常组和高磷组(给予 10 mmol / L β-甘油磷酸盐刺激)。 采用实时荧光定量 PCR 检测各组 VSMCs miR-30b 表达、Western blot 法测定促凋亡基因 BAX、抑凋亡基因 Bcl-2 的表达情况,采用 MTT 法检测各组细胞的增殖情况、采用流式细胞法检测各组细胞的凋亡情况。 为进一步验证 miR-30b 对 VSMCs 凋亡的影响,给与转染 miR-30b 的抑制物 inhibitor-30b 和类似物 mimic-30b,观察 VSMCs 增殖和凋亡情况以及 BAX、Bcl-2 表达情况。 结果 (1)高磷对 VSMCs 凋亡的影响:流式结果显示,高磷组 VSMCs 凋亡细胞显著增多(P<0.05);MTT结果显示,高磷组细胞增殖减少(P<0. 05);高磷组 BAX 表达升高、Bcl-2 表达降低(P<0.05);高磷组 miR-30b 的表达显著下降(P<0. 05)。 (2)miR-30b 对 VSMCs 凋亡的影响:转染 inhibitor-30b 后 VSMCs 凋亡细胞显著增多(P<0. 05);转染 inhibitor-30b 后 MTT 显示细胞增殖减少(P<0.05);转染 inhibitor-30b 后,BAX 表达升高、Bcl-2 表达降低(P<0.05);转染 mimic-30b 后 VSMCs 凋亡细胞显著减少 (P<0.05);转染 mimic-30b 后,BAX 表达降低、Bcl-2 表达升高(P<0.05)。 结论 高磷可以诱导细胞凋亡,其可能机制之一是高磷通过抑制 miRNA-30b 表达,进而促进凋亡基因 BAX 表达,降低抑凋亡基因 Bcl-2 表达,导致 VSMCs 发生凋亡。

    Abstract:

    Objective To observe the role and mechanism of miR-30b in apoptosis of vascular smooth muscle cells induced by high phosphorus. Methods VSMCs from the rat thoracic aorta were cultured in vitro and divided into a normal group and high phosphorus group stimulated by 10 mmol / L β-glycerophosphate. Expression of miR-30b in VSMCs was quantified by Real-time PCR, and expression of pro-apoptotic protein BAX and anti-apoptotic protein Bcl-2 was determined by Western blot. Cell proliferation was assessed by MTT assays, and apoptosis was detected by flow cytometry. To verify the effect of miR-30b on VSMCs apoptosis, inhibitor-30b and mimic-30b were applied to observe changes in proliferation and apoptosis of VSMCs and expression of BAX and Bcl-2. Results (1)The effect of high phosphorus on apoptosis of VSMCs: Flow cytometry showed that apoptosis cells of VSMCs in high phosphorus group increased significantly (P<0.05). The results of MTT showed that the proliferation of cells in high phosphorus group decreased ( P< 0.05). The expression of BAX increased and Bcl-2 decreased in high phosphorus group (P<0.05). The expression of miR-30b in high phosphorus group decreased significantly (P<0.05). (2) Effect of miR-30b on apoptosis of VSMCs: After transfection of inhibitor-30b, the number of apoptotic cells in VSMCs increased significantly (P<0.05). After transfection of inhibitor-30b, MTT assay showed that cell proliferation decreased (P<0.05). The expression of BAX increased and the expression of Bcl-2 decreased after transfection of inhibitor-30b (P<0.05). After mimic-30b transfection, the apoptotic cells of VSMCs decreased significantly (P<0.05). After transfection of mimic-30b, the expression of BAX decreased and the expression of Bcl-2 increased (P<0.05). Conclusions High phosphorus induces apoptosis. A possible mechanism is that high phosphorus inhibits miR-30b expression and then promotes expression of apoptosis gene BAX and decreases expression of anti-apoptosis gene Bcl-2, which leads to VSMCs apoptosis.

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刘 兰,张东雪,张胜雷,白亚玲. miR-30b 在高磷诱导的血管平滑肌细胞凋亡中的作用[J].中国比较医学杂志,2022,32(7):58~65.

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  • 收稿日期:2021-09-27
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  • 在线发布日期: 2022-11-09
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