Chen Lu-Ping, Yang Yi-Tong, Zhao Miao-Miao, Li Han-Wen, Sun Wen-Ting, Shi Zhao-Ling
Second Affiliated Hospital of Shaanxi University of Chinese Medicine, Xianyang, Shaanxi 712000, China.
Zhongguo Dang Dai Er Ke Za Zhi. 2024;26(9):967-973. doi: 10.7499/j.issn.1008-8830.2405084.
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).
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.
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 (<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 (<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 (<0.05).
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.
研究川芎嗪（TMP）对肿瘤坏死因子-α（TNF-α）诱导的人冠状动脉内皮细胞（HCAEC）炎症损伤的影响及机制。
将HCAEC随机分为四组：对照组（未处理）、模型组（用50 ng/mL TNF-α处理24小时）、TMP组（先用80 μg/mL TMP预处理12小时，然后用TNF-α处理24小时）和SIRT1抑制剂组（先用TMP和特异性SIRT1抑制剂EX527预处理12小时，然后用TNF-α处理24小时）。采用CCK-8法评估细胞活力，用乳酸脱氢酶（LDH）检测试剂盒检测LDH活性，用DCFH-DA染色观察活性氧（ROS）水平，用蛋白质免疫印迹法检测焦亡相关蛋白的表达，用免疫荧光染色分析SIRT1表达。
与对照组相比，模型组细胞活力降低，LDH活性、ROS水平和焦亡相关蛋白表达增加，SIRT1表达降低（<0.05）。与模型组相比，TMP组细胞活力增加，LDH活性、ROS水平和焦亡相关蛋白表达降低，SIRT1表达增加（<0.05）。与TMP组相比，SIRT1抑制剂组细胞活力降低，LDH活性、ROS水平和焦亡相关蛋白表达增加，SIRT1表达降低（<0.05）。
TMP可能减轻TNF-α诱导的HCAEC炎症损伤，这与抑制焦亡和激活SIRT1信号通路有关。
