PLD2 通过 Nrf2/NF-κB 通路调节胰腺细胞凋亡和细胞水肿的分子机制。
The molecular mechanism of PLD2-mediated regulation of apoptosis and cell edema in pancreatic cells via the Nrf2/NF-κB pathway.
机构信息
Department of General Surgery, Wuhan Fourth Hospital, No. 435 Guli Road, Dongxihu District, Wuhan, 430000, Hubei, China.
出版信息
Sci Rep. 2024 Oct 26;14(1):25563. doi: 10.1038/s41598-024-76274-4.
This study aimed to elucidate the molecular mechanisms by which PLD2 controls apoptosis and edema in pancreatic cells via the Nrf2/NF-κB pathway. AR42J rat pancreatic cells were treated with 10 nM mitomycin to create an in vitro pancreatitis model (model group), with a control group receiving phosphate-buffered saline. Cells were transfected with a PLD2 overexpression plasmid using Lipofectamine 3000, forming the PLD2 overexpression group. PLD2 protein expression was assessed by Western blotting, and TNF-α, IL-6, and IL-10 levels were measured by RT-qPCR. Nrf2/NF-κB protein expressions were also analyzed. Apoptosis and necrosis were evaluated using Annexin V-FITC/PI staining and the LDH release test. Cell edema was assessed by cell volume, ion content, and membrane damage. Western blotting was used to analyze pan-apoptosis-related proteins. PLD2 expression was lower in the model group compared to controls (P < 0.05) but higher in the PLD2 overexpression group (P < 0.05). TNF-α, IL-6, and IL-10 levels were elevated in the model group (P < 0.05) and reduced in the PLD2 overexpression group (P < 0.05). Nrf2 expression decreased in the model group but increased with PLD2 overexpression (P < 0.05). NF-κB expression increased in the model group but decreased with PLD2 overexpression (P < 0.05). Apoptosis and necrosis rates were higher in the model group (P < 0.05) but lower in the PLD2 overexpression group (P < 0.05). Cell volume, Na + content, and LDH release increased in the model group (P < 0.05) but decreased with PLD2 overexpression (P < 0.05). RIPK1 expression decreased in the model group (P < 0.05) but increased with PLD2 overexpression (P < 0.05). CASP8, FADD, and ZBP1 levels were higher in the model group (P < 0.05) and reduced with PLD2 overexpression (P < 0.05). PLD2 exerts a protective effect in acute pancreatitis by activating Nrf2 and inhibiting NF-κB, reducing apoptosis, cell swelling, and membrane damage. This highlights potential therapeutic targets for pancreatic inflammation.
本研究旨在阐明 PLD2 通过 Nrf2/NF-κB 通路控制胰腺细胞凋亡和水肿的分子机制。使用 10 nM 丝裂霉素处理 AR42J 大鼠胰腺细胞以建立体外胰腺炎模型（模型组），对照组给予磷酸盐缓冲液。使用 Lipofectamine 3000 将 PLD2 过表达质粒转染细胞，形成 PLD2 过表达组。通过 Western blot 评估 PLD2 蛋白表达，通过 RT-qPCR 测量 TNF-α、IL-6 和 IL-10 水平。还分析了 Nrf2/NF-κB 蛋白表达。通过 Annexin V-FITC/PI 染色和 LDH 释放试验评估细胞凋亡和坏死。通过细胞体积、离子含量和膜损伤评估细胞水肿。通过 Western blot 分析全凋亡相关蛋白。与对照组相比，模型组中 PLD2 表达降低（P<0.05），但在 PLD2 过表达组中表达升高（P<0.05）。模型组中 TNF-α、IL-6 和 IL-10 水平升高（P<0.05），PLD2 过表达组中降低（P<0.05）。模型组中 Nrf2 表达降低，但 PLD2 过表达后增加（P<0.05）。模型组中 NF-κB 表达增加，但 PLD2 过表达后降低（P<0.05）。模型组中细胞凋亡和坏死率较高（P<0.05），但 PLD2 过表达组中较低（P<0.05）。模型组中细胞体积、Na+含量和 LDH 释放增加（P<0.05），但 PLD2 过表达后降低（P<0.05）。模型组中 RIPK1 表达降低（P<0.05），但 PLD2 过表达后增加（P<0.05）。模型组中 CASP8、FADD 和 ZBP1 水平升高（P<0.05），PLD2 过表达后降低（P<0.05）。PLD2 通过激活 Nrf2 和抑制 NF-κB，减少细胞凋亡、肿胀和膜损伤，对急性胰腺炎发挥保护作用。这突出了胰腺炎症的潜在治疗靶点。
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