羌活内酯 I 通过抗氧化、抗炎和抗细胞凋亡途径减轻小鼠肝缺血/再灌注损伤。
Senkyunolide I attenuates hepatic ischemia/reperfusion injury in mice via anti-oxidative, anti-inflammatory and anti-apoptotic pathways.
机构信息
Jiangsu Province Key Laboratory of Anesthesiology, Xuzhou Medical University, Xuzhou, China; Faculty of Anesthesiology, Changhai Hospital, Naval Medical University, Shanghai, China.
Faculty of Anesthesiology, Changhai Hospital, Naval Medical University, Shanghai, China.
出版信息
Int Immunopharmacol. 2021 Aug;97:107717. doi: 10.1016/j.intimp.2021.107717. Epub 2021 Apr 29.
BACKGROUND
Senkyunolide I (SEI)exerts considerable protective effects in various disease models, but its effect on hepatic ischemia-reperfusion (I/R) injury remains unknown. This research aimed to investigate the effect of SEI in a murine model of hepatic I/R injury.
METHODS
With modified liver I/R murine model, low, medium and high doses of SEI were injected intraperitoneally after operation. After 6 h of reperfusion, the blood and liver were collected. Serum ALT and AST were detected by automatic analyzer, while liver injury was evaluated by HE staining. High-dose SEI was selected to further explore its impacts on oxidative stress, inflammatory responses and apoptosis induced by hepatic I/R. The pharmacological effect of SEI was also compared with a positive control, glutathione (GSH). We used ELISA to detect serum TNF-α, IL-1 β and IL-6, special kit to explore activities of SOD and GSH-Px, and the content of MDA, and western blotting to detect HO-1, Bax and Bcl-2 levels, and to perceive expressions and phosphorylations of NF- κB p65 and p38/ERK/JNK in liver tissues. Apoptosis in liver tissue was evaluated by TUNEL. The antioxidative effect of SEI was further investigated using the HuCCT1 cells stimulated with HO and the role of SEI on regulation of Nrf-2/HO-1 was determined.
RESULTS
200 mg/kg of SEI was optimal dose for treating liver I/R injury. Elevated ALT, AST and histopathological injury in I/R liver was attenuated by SEI administration, similarly to GSH. Serum TNF-α, IL-1β, and IL-6 were reduced in liver I/R mice treated with SEI, and in liver tissues, phosphorylation of p65 NF-κB and MAPK kinases (p38, ERK, JNK), were inhibited. SEI reduced the MDA content, but increased HO-1 level and enhanced SOD and GSH-Px activities. Apoptosis of liver tissues was decreased, while SEI inhibited Bax and elevated Bcl-2 expression. In in vitro experiments, HO reduced the survival rate of HuCCT1 cells, which was protected by SEI administration. SEI reduced the ROS and MDA content. The transportation of Nrf-2 into the nucleus was enhanced and HO-1 expression was upregulated.
CONCLUSIONS
SEI attenuates hepatic I/R injury in mice via anti-oxidative, anti-inflammatory and anti-apoptotic pathways.
背景
千里光叶内酯 I（SEI）在多种疾病模型中表现出相当大的保护作用，但它对肝缺血再灌注（I/R）损伤的影响尚不清楚。本研究旨在探讨 SEI 在肝 I/R 损伤小鼠模型中的作用。
方法
采用改良的肝 I/R 小鼠模型，在手术后经腹腔注射低、中、高剂量 SEI。再灌注 6 小时后，采集血液和肝脏。采用自动分析仪检测血清 ALT 和 AST，通过 HE 染色评估肝损伤。选择高剂量 SEI 进一步探讨其对肝 I/R 诱导的氧化应激、炎症反应和细胞凋亡的影响。并将 SEI 的药理作用与阳性对照物谷胱甘肽（GSH）进行比较。采用 ELISA 法检测血清 TNF-α、IL-1β和 IL-6，特殊试剂盒检测 SOD 和 GSH-Px 的活性以及 MDA 的含量，Western blot 检测肝组织中 HO-1、Bax 和 Bcl-2 的表达和磷酸化水平，以及 NF-κB p65 和 p38/ERK/JNK 的表达和磷酸化水平，并通过 TUNEL 评估肝组织中的细胞凋亡。进一步采用 HO 刺激 HuCCT1 细胞探讨 SEI 的抗氧化作用，并确定 SEI 对 Nrf-2/HO-1 调节的作用。
结果
200mg/kg 的 SEI 是治疗肝 I/R 损伤的最佳剂量。SEI 给药可减轻 I/R 肝中 ALT、AST 的升高和组织病理学损伤，与 GSH 作用相似。SEI 治疗可降低肝 I/R 小鼠的血清 TNF-α、IL-1β和 IL-6，并抑制肝组织中 p65 NF-κB 和 MAPK 激酶（p38、ERK、JNK）的磷酸化。SEI 降低 MDA 含量，增加 HO-1 水平，增强 SOD 和 GSH-Px 的活性。降低肝组织细胞凋亡，抑制 Bax 表达，上调 Bcl-2 表达。在体外实验中，HO 降低 HuCCT1 细胞的存活率，而 SEI 给药可保护细胞。SEI 降低 ROS 和 MDA 含量，增强 Nrf-2 向核内转运，上调 HO-1 表达。
结论
SEI 通过抗氧化、抗炎和抗细胞凋亡途径减轻小鼠肝 I/R 损伤。
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