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| 利多卡因对糖尿病大鼠脊髓神经元的损伤作用 |
| Injury effect of lidocaine on spinal cord neurons of diabetic rats |
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| DOI：10.12089/jca.2023.03.013 |
| 中文关键词: 利多卡因 糖尿病 脊髓神经元 损伤 |
| 英文关键词: Lidocaine Diabetes mellitus Spinal cord neurons Injury |
| 基金项目:江西省自然科学基金资助项目（20192BAB205041）;江西省教育厅科学技术研究项目（180135） |
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| 中文摘要: |
目的 探讨利多卡因对糖尿病大鼠脊髓神经元的损伤作用。
方法 选择健康SPF级雄性SD大鼠30只、GK大鼠30只，12周龄，体重180～200 g。取大鼠脊髓神经元进行体外分离培养。采用随机数字表法将大鼠脊髓神经元分为四组：SD大鼠脊髓神经元组（C组）、SD大鼠脊髓神经元+利多卡因组（CL组）、GK大鼠脊髓神经元组（GK组）和GK大鼠脊髓神经元+利多卡因组（GKL组）。C组和GK组置于DMEM高糖培养基中，在37℃，5%CO2O培养箱内培养24 h，CL组和GKL组在相同条件下加入浓度为2 mmol/L的利多卡因培养24 h。采用Western blot法检测脊髓神经元中自噬相关蛋白PINK1、mTORC1、AMPK、p-AMPK、LC3Ⅱ和LC3Ⅰ含量，DCFH-DA法检测脊髓神经元中活性氧（ROS）含量，免疫荧光法检测脊髓神经元中PINK1和LC3含量。
结果 与C组比较，CL组、GK组和GKL组脊髓神经元PINK1、LC3含量和p-AMPK/AMPK、LC3Ⅱ/Ⅰ比值均明显升高（P<0.05），mTORC1含量明显降低（P<0.05），ROS含量明显升高（P<0.05）。与GK组比较，GKL组脊髓神经元PINK1、LC3含量和p-AMPK/AMPK、LC3Ⅱ/Ⅰ比值明显升高（P<0.05），mTORC1含量明显降低（P<0.05），ROS含量明显升高（P<0.05）。
结论 利多卡因诱导GK大鼠脊髓神经元的自噬相关蛋白的表达及ROS的释放，加重对GK大鼠脊髓神经元的损伤。 |
| 英文摘要: |
Ojective To explore the injury effect of lidocaine in spinal cord neurons of diabetic rats.
Methods Thirty SPF healthy SD rats and thirty GK rats, aged 12 weeks, weighing 180-200 g were selected. The spinal cord neurons of rats were isolated and cultured in vitro, and randomly divided into four groups: spinal cord neurons of SD rats group (group C), spinal cord neurons of SD rats + lidocaine group(group CL), spinal cord neurons of GK rats group(group GK), and spinal cord neurons of GK rats + lidocaine group (group GKL). Spinal cord neurons in groups C and GK were placed in DMEM high glucose medium and cultured in an incubator with 5% CO2O at 37 ℃ for 24 hours. Lidocaine at a concentration of 2 mmol/L was added to groups CL and GKL, and then neurons in groups CL and GKL were cultured for 24 hours under the same conditions as groups C and GK. The content of autophagy-related proteins PINK1, mTORC1, AMPK, p-AMPK, LC3Ⅱ, and LC3Ⅰ in spinal cord neurons was measured by Western blot. The content of reactive oxygen species (ROS) in spinal cord neurons was detected by DCFH-DA. The content of PINK1 and LC3 in spinal cord neurons was measured by immunofluorescence.
Results Compared with group C, the content of PINK1, LC3, and p-AMPK/AMPK, LC3Ⅱ/Ⅰ level in groups CL, GK and GKL were significantly increased (P < 0.05), the content of mTORC1 was significantly decreased (P < 0.05), and the content of ROS was significantly increased (P < 0.05). Compared with group GK, the content of PINK1, LC3, and p-AMPK/AMPK,LC3Ⅱ/Ⅰ level in the group GKL were significantly increased (P < 0.05), the content of mTORC1 was significantly decreased (P < 0.05), and the content of ROS was significantly increased (P < 0.05).
Conclusion Lidocaine aggravates the damage to spinal cord neurons of GK rats by promoting the expression of autophagy-related proteins and the release of ROS. |
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