基础医学与临床 ›› 2023, Vol. 43 ›› Issue (1): 137-143.doi: 10.16352/j.issn.1001-6325.2023.01.0137

• 研究论文 • 上一篇    下一篇

丙泊酚减轻低氧诱导的大鼠肾上腺嗜铬细胞瘤细胞系PC12炎性反应和细胞凋亡

顾炜, 祝剑平, 吴品雯*   

  1. 复旦大学附属闵行医院 麻醉科,上海 201199
  • 收稿日期:2022-01-07 修回日期:2022-05-31 发布日期:2022-12-27
  • 通讯作者: *wpw1976@qq.com
  • 基金资助:
    上海市闵行区中心医院院级课题(2020MHLC01)

Propofol attenuates hypoxia-induced inflammation and apoptosis in rat pheochromocytoma cell line PC12

GU Wei, ZHU Jianping, WU Pinwen*   

  1. Department of Anesthesiology, Minhang Hospital Affiliated to Fudan University, Shanghai 201199, China
  • Received:2022-01-07 Revised:2022-05-31 Published:2022-12-27
  • Contact: *wpw1976@qq.com

摘要: 目的 研究丙泊酚是否通过抑制miR-141-3p的表达,减轻低氧诱导的肾上腺嗜铬细胞瘤细胞系PC12炎性反应和细胞凋亡的分子机制。方法 将PC12细胞分为对照组、低氧组、(5、10、20)μmol/L丙泊酚+低氧组、anti-miR-con+低氧组、anti-miR-141-3p+低氧组、miR-con+20 μmol/L丙泊酚+低氧组、miR-141-3p+20 μmol/L丙泊酚+低氧组。流式细胞仪检测PC12细胞的凋亡;Western blot检测活化的含半胱氨酸天冬氨酸蛋白水解酶-3(cleaved caspase-3)蛋白表达;相关试剂盒检测丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性;活性氧荧光探针DCFH-DA法测定活性氧(ROS)含量;ELISA检测肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-6含量;RT-qPCR检测miR-141-3p表达。结果 与对照组比较,低氧组PC12细胞的凋亡率、cleaved caspase-3蛋白表达水平、MDA、ROS、TNF-α、IL-1β、IL-6含量和miR-141-3p表达量均升高,SOD活性减弱(P<0.05)。与低氧组比较,5、10、20 μmol/L丙泊酚+低氧组PC12细胞的凋亡率、cleaved caspase-3蛋白表达水平、MDA含量、ROS、TNF-α、IL-1β、IL-6含量和miR-141-3p表达量均随丙泊酚浓度的增加呈降低趋势,SOD活性随丙泊酚浓度的增加呈升高趋势(P<0.05)。Anti-miR-141-3p+低氧组PC12细胞的miR-141-3p表达量、cleaved caspase-3蛋白表达水平、MDA、ROS、TNF-α、IL-1β、IL-6含量和细胞凋亡率均比anti-miR-con+低氧组降低,SOD活性比anti-miR-con+低氧组增加(P<0.05)。miR-141-3p+20 μmol/L丙泊酚+低氧组PC12细胞的miR-141-3p表达量、cleaved caspase-3蛋白表达水平、MDA含量、ROS、TNF-α、IL-1β、IL-6含量和细胞凋亡率均高于miR-con+20 μmol/L 丙泊酚+低氧组,SOD活性低于miR-con+20 μmol/L 丙泊酚+低氧组(P<0.05)。结论 丙泊酚可能通过抑制miR-141-3p的表达,减轻低氧诱导的PC12细胞炎性反应、氧化应激和细胞凋亡。

关键词: PC12细胞, 炎性反应, 氧化应激, 凋亡, miR-141-3p

Abstract: Objective To investigate whether propofol inhibits the expression of miR-141-3p and reduces the molecular mechanism of hypoxia-induced inflammation and apoptosis of pheochromocytoma cell line PC12. Methods PC12 cells were divided into control group, hypoxia group, (5, 10, 20)μmol/L propofol+hypoxia group, anti-miR-con+hypoxia group, anti-miR-141-3p+hypoxia group, miR-con+20 μmol/L propofol+hypoxia group, miR-141-3p+20 μmol/L propofol+hypoxia group. Flow cytometry was used to detect the apoptosis of PC12 cells; Western blot was employed to determine the expression of activated cleaved caspase-3 (cleaved caspase-3) protein, and the kits were implemented to monitor malondialdehyde (MDA) content and superoxide dismutase (SOD) activity; Reactive oxygen species fluorescent probe DCFH-DA method to determine reactive oxygen species (ROS) content; ELISA kits to assay tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 content, RT-qPCR to detect the expression of miR-141-3p. Results Compared with the control group, the apoptosis rate, cleaved caspase-3 protein expression level, MDA, ROS, TNF-α, IL-1β, IL-6 content and miR-141-3p expression of PC12 cells in hypoxia group were all increased, while SOD activity weakened (P<0.05). Compared with the hypoxia group, the apoptosis rate, cleaved caspase-3 protein expression level, MDA content, ROS, TNF-α, IL-1β, IL-6 content and miR-141-3p expression of PC12 cells in 5, 10, 20 μmol/L propofol+hypoxia group decreased with the increase of propofol concentration, and SOD activity increased with the increase of propofol concentration (P<0.05). The expression of miR-141-3p, cleaved caspase-3 protein expression, MDA, ROS, TNF-α, IL-1β, IL-6 content and apoptosis rates of PC12 cells in the anti-miR-141-3p+ hypoxia group were lower than those in the anti-miR-con+hypoxia group, and the SOD activity was higher than that in the anti-miR-con+hypoxia group (P<0.05). The miR-141-3p expression, cleaved caspase-3 protein expression level, MDA content, ROS, TNF-α, IL-1β, IL-6 content and cell apoptosis rate of PC12 cells in miR-141-3p+20 μmol/L propofol+hypoxia group were higher than miR-con+20 μmol/L propofol+hypoxia group, while SOD activity was lower than those from miR-con+20 μmol/L propofol+hypoxia group (P<0.05). Conclusions Propofol can alleviate the inflammatory response, oxidative stress and apoptosis of PC12 cells induced by hypoxia by inhibiting the expression of miR-141-3p.

Key words: PC12 cells, inflammatory response, oxidative stress, apoptosis, miR-141-3p

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