目的 观察川芎嗪（tetramethylpyrazine, TMP）对肿瘤坏死因子-α（tumor necrosis factor-α, TNF-α）诱导的人冠状动脉内皮细胞（human coronary artery endothelial cell, HCAEC）炎症损伤的作用及机制。 方法 将HCAEC随机分为4组：对照组不给予任何处理，模型组给予TNF-α（50 ng/mL）处理24 h，TMP组给予TMP（80 μg/mL）预处理12 h后使用TNF-α（50 ng/mL）处理24 h，沉默信息调节因子1（silent information regulaton 1, SIRT1）抑制剂组给予TMP（80 μg/mL）及SIRT1特异性抑制剂EX527预处理12 h后使用TNF-α（50 ng/mL）处理24 h。CCK-8法检测各组细胞活力，乳酸脱氢酶（lactate dehydrogenase, LDH）试剂盒检测各组细胞培养基中LDH活性，DCFH-DA染色法观察细胞活性氧（reactive oxygen species, ROS）含量变化，Western blot法检测焦亡相关蛋白表达，免疫荧光染色检测各组细胞SIRT1表达。 结果 与对照组相比，模型组细胞活力降低，LDH活性、ROS水平、焦亡相关蛋白表达升高，SIRT1表达降低（P<0.05）；与模型组相比，TMP组细胞活力升高，LDH活性、ROS水平、焦亡相关蛋白表达降低，SIRT1表达升高（P<0.05）；与TMP组比较，SIRT1抑制剂组细胞活力下降，LDH活性、ROS水平、焦亡相关蛋白表达升高，SIRT1表达降低（P<0.05）。 结论 TMP可减轻TNF-α诱导的HCAEC炎症损伤，与抑制细胞焦亡、激活SIRT1信号有关。
Abstract
Objective To study the effects and mechanisms of tetramethylpyrazine (TMP) on tumor necrosis factor-α (TNF-α)-induced inflammatory injury in human coronary artery endothelial cells (HCAEC). Methods HCAEC were randomly divided into four groups: the control group (no treatment), the model group (treated with TNF-α, 50 ng/mL for 24 hours), the TMP group (pre-treated with TMP, 80 μg/mL for 12 hours followed by TNF-α treatment for 24 hours), and the SIRT1 inhibitor group (pre-treated with TMP and the specific SIRT1 inhibitor EX527 for 12 hours followed by TNF-α treatment for 24 hours). Cell viability was assessed using the CCK-8 method, lactate dehydrogenase (LDH) activity was measured using an LDH assay kit, reactive oxygen species (ROS) levels were observed using DCFH-DA staining, expression of pyroptosis-related proteins was detected by Western blot, and SIRT1 expression was analyzed using immunofluorescence staining. Results Compared to the control group, the model group showed decreased cell viability, increased LDH activity, ROS level and expression of pyroptosis-related proteins, and decreased SIRT1 expression (P<0.05). Compared to the model group, the TMP group exhibited increased cell viability, decreased LDH activity, ROS level and expression of pyroptosis-related proteins, and increased SIRT1 expression (P<0.05). In comparison to the TMP group, the SIRT1 inhibitor group showed decreased cell viability, increased LDH activity, ROS level and expression of pyroptosis-related proteins, and decreased SIRT1 expression (P<0.05). Conclusions TMP may attenuate TNF-α-induced inflammatory injury in HCAEC, which is associated with the inhibition of pyroptosis and activation of the SIRT1 signaling pathway.
关键词
炎症损伤 /
川芎嗪 /
SIRT1 /
焦亡 /
人冠状动脉内皮细胞
Key words
Inflammatory injury /
Tetramethylpyrazine /
SIRT1 /
Pyroptosis /
Human coronary artery endothelial cell
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参考文献
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基金
国家自然科学基金青年科学基金项目（82205188）；陕西省重点研发计划项目（2023-YBSF-638）。
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