miR-214对异丙酚麻醉大鼠术后神经元损伤的保护作用
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乐山市人民医院麻醉科,四川 乐山 614000

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R-33


Protective effect of miR-214 on neuronal injury in propofol-anesthetized thoracotomy rats
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Department of Anesthesiology, Leshan People’s Hospital, Leshan 614000, China

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    摘要:

    目的 探讨 miR-214 对异丙酚麻醉诱导的神经元损伤的保护作用及生物学机制。 方法 7 日龄 SD雄性大鼠随机分为 4 组(每组 15 只):生理盐水组(NS)、异丙酚麻醉开腹探查组(模型)、miR-NC 组和 miR-214 组。除 NS 组外,其余组别建立异丙酚麻醉开腹探查模型。miR-NC 组和 miR-214 组分别在麻醉前将 miR-NC-agomir 或miR-214-agomir 注射到海马内。采用免疫荧光法检测海马 mTORC1 的表达,TUNEL 染色检测海马组织细胞凋亡,RT-qPCR 分析海马组织中 miR-214 表达。 分别将 miR-214 抑制剂和 mTORC1 抑制剂转染入 HT22 海马神经元细胞,然后暴露于异丙酚。 采用流式细胞术分析细胞的存活情况和 Western blot 分析 TEFB、C-caspase3 蛋白表达。 结果 与 NS 组相比,模型组大鼠海马中 miR-214 明显下调(P<0. 05),并且海马神经细胞凋亡显著增加(P<0. 05)。与模型组相比,miR-214 组海马神经元凋亡减弱(P<0. 05)。 生物信息学预测证明 mTORC1 和 miR-214 之间存在特异性结合位点。 免疫荧光检测显示,与 NS 组比较,模型组诱导海马神经组织中 mTORC1 表达增加(P<0. 05),miR-214 治疗显著降低了 mTORC1 表达(P<0. 05)。 此外,与 NC 对照组相比,si-mTORC1 转染导致暴露于异丙酚的HT22 海马神经元细胞的凋亡率显著降低,并且 TFEB 表达显著增加(P<0. 01),以及 C-caspase3 降低(P<0. 05)。而 miR-214 抑制剂转染显著逆转了 si-mTORC1 的保护作用以及诱导的 TFEB、C-caspase3 蛋白表达的变化( P<0. 05)。 结论 miR-214 可能通过 mTORC1-TFEB 通路减轻异丙酚神经毒性,提高神经元的存活率。

    Abstract:

    Objective To investigate the protective effect and biological mechanism of miR-214 on neuronal injury induced by propofol anesthesia, and elucidate the underlying mechanisms of this process. Methods Seven-day-old male Sprague-Dawley rats were randomly divided into four groups (15 rats per group): normal saline (NS), propofol anesthesia thoracotomy exploration ( model), miR-NC and miR-214. With the exception of the NS group, all groups underwent establishment of the propofol anesthesia thoracotomy exploration model. In miR-NC and miR-214 groups, miR-NC-agomir or miR-214-agomir, respectively, were injected into hippocampus before anesthesia. Expression of mTORC1 in the hippocampus was detected by immunofluorescence, apoptosis was detected by TUNEL staining, and expression of miR-214 in hippocampus was analyzed by RT-qPCR. HT22 hippocampal neurons were transfected with an miR-214 inhibitor and mTORC1 inhibitor, and then exposed to propofol. Flow cytometry was used to analyze cell survival, whereas Western blot was used to analyze protein expression of TEFB and C-caspase3. Results Compared with the NS group, miR-214 in the hippocampus of the model group was significantly downregulated (P< 0.05) and apoptosis of hippocampal neurons was significantly increased (P< 0.05). Compared with the model group, apoptosis of hippocampal neurons in the miR-214 group was decreased ( P< 0.05). Bioinformatics prediction indicated the presence of a specific binding site between mTORC1 and miR-214. Compared with the NS group, expression of mTORC1 was increased in the model group ( P<0. 05), and miR-214 treatment significantly reduced expression of mTORC1 (P<0.05). In addition, compared with the NC group, si-mTORC1 transfection significantly reduced the apoptosis rate of HT22 hippocampal neurons exposed to propofol (P< 0.05), increased TFEB expression (P< 0.01), and decreased cleaved caspase 3 ( P< 0. 05). miR-214 inhibitor transfection significantly reversed the protective effect of si-mTORC1 and changes of TFEB and C-caspase3 protein expression induced by si-mTORC1 ( P< 0.05). Conclusions miR-214 attenuates propofol neurotoxicity through the mTORC1-TFEB pathway and improves the survival rate of neurons.

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杨继梅,卢文江,周娇洁. miR-214对异丙酚麻醉大鼠术后神经元损伤的保护作用[J].中国比较医学杂志,2022,32(9):39~46,81.

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  • 收稿日期:2021-11-14
  • 在线发布日期: 2023-01-16
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