去甲汉黄芩素通过激活Nrf2/HO-1信号通路缓解急性低压低氧诱导肺组织损伤的作用

景临林; 邹蓓蕾; 辛宇; 马慧萍

doi:10.13748/j.cnki.issn1007-7693.20250136

去甲汉黄芩素通过激活Nrf2/HO-1信号通路缓解急性低压低氧诱导肺组织损伤的作用

Effect of Norwogonin in Alleviating Acute Hypobaric Hypoxia Induced Lung Tissue Injury by Activating Nrf2/HO-1 Signaling Pathway

摘要

摘要:
目的研究去甲汉黄芩素改善急性低压低氧(acute hypobaric hypoxia，AHH)诱导小鼠肺组织损伤的作用与潜在机制。
方法 78只雄性BALB/c小鼠随机分为正常对照组、AHH组、芦丁组和去甲汉黄芩素低、中、高剂量组6组。正常对照组、AHH组小鼠腹腔注射生理盐水，4组给药组分别腹腔注射芦丁(200 mg·kg⁻¹)和低、中、高剂量(50、100、200 mg·kg⁻¹)去甲汉黄芩素。将小鼠置于模拟海拔8 000 m的动物实验舱暴露24 h，构建AHH肺损伤模型。HE染色观察肺组织病理变化；试剂盒检测肺组织中过氧化氢(H₂O₂)、丙二醛(malondialdehyde，MDA)、超氧化物歧化酶(superoxide dismutase，SOD)和谷胱甘肽(glutathione，GSH)的水平以及肿瘤坏死因子α(tumor necrosis factor-α，TNF-α)、白细胞介素1β(interleukin-1β，IL-1β)和白细胞介素6(interleukin-6，IL-6)的含量；Western blotting检测肺组织中缺氧相关蛋白缺氧诱导因子-1α(hypoxia-inducible factor-1α，HIF-1α)和血管内皮生长因子(vascular endothelial growth factor，VEGF)、抗氧化应激相关蛋白核因子-E2相关因子2(nuclear factor erythroid 2-related factor 2，Nrf2)和血红素加氧酶-1(heme oxygenase-1，HO-1)、炎性相关蛋白核因子κB(nuclear factor-κB，NF-κB)和TNF-α以及凋亡相关蛋白B淋巴细胞瘤-2(B-cell lymphoma-2，Bcl-2)、Bcl-2关联X蛋白(Bcl-2-associated X protein，Bax)和裂解的半胱氨酸-天冬氨酸蛋白酶-3(cleaved cysteine-aspartic acid caspase-3，Cleaved caspase-3)的表达水平；AutoDock软件进行分子对接。
结果与AHH组相比，去甲汉黄芩素处理能显著减少肺组织含水量和病理学变化，降低肺组织中H₂O₂、MDA、IL-1β、TNF-α和IL-6的含量，升高SOD活力和GSH水平，下调HIF-1α、VEGF、NF-κB、TNF-α、Bax和Cleaved caspase-3蛋白表达，上调Nrf2、HO-1、Bcl-2蛋白表达，减少Bax/Bcl-2的比值。分子对接结果显示，去甲汉黄芩素与Nrf2、HO-1表现出良好的结合能力。
结论去甲汉黄芩素通过激活Nrf2/HO-1信号通路缓解氧化应激，减少炎性反应，并抑制细胞凋亡，从而改善AHH诱导的小鼠肺组织损伤。

Abstract:
OBJECTIVE To investigate the protective effects and potential mechanisms of norwogonin in ameliorating acute hypobaric hypoxia(AHH) induced lung tissue injury in mice.
METHODS Seventy-eight male BALB/c mice were randomly divided into 6 groups: the normal control group, the AHH group, the rutin group, and the low-, medium-, and high-dose groups of norwogonin. Mice in the normal control group and AHH group were intraperitoneally injected with normal saline, and the administration groups were intraperitoneally injected with rutin(200 mg·kg⁻¹) and low-, medium-, and high-dose of norwogonin(50, 100, 200 mg·kg⁻¹). The mouse model of AHH lung injury was established by exposure to an altitude of 8000 m animal experimentation chamber for 24 h. Pathological changes in mice lungs were evaluated using HE staining. The levels of H₂O₂, malondialdehyde(MDA), superoxide dismutase(SOD), and glutathione(GSH) and the contents of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), and interleukin-6(IL-6) in lung tissue were detected using commercial kits. The levels of hypoxia-related proteinshypoxia-inducible factor-1α(HIF-1α) and vascular endothelial growth factor(VEGF), antioxidative stress-related proteinsnuclear factor erythroid 2-related factor 2(Nrf2) and heme oxygenase-1(HO-1), inflammation-related proteinsnuclear factor-κB(NF-κB) and TNF-α, and apoptosis-related proteinsB-cell lymphoma-2(Bcl-2), Bcl-2-associated X protein(Bax), and cleaved cysteine-aspartic acid caspase-3(Cleaved caspase-3) in lung tissues were detected by Western blotting. AutoDock software was used to perform molecular docking.
RESULTS Compared to the AHH group, norwogonin administration remarkably decreased the water content and pathological changes in lung tissues, reduced the levels of H₂O₂, MDA, IL-1β, TNF-α and IL-6, and elevated SOD activity and GSH level, downregulated the expression levels of HIF-1α, VEGF, NF-κB, TNF-α, Bax and Cleaved caspase-3 proteins, upregulated the Nrf2, HO-1, Bcl-2 protein expression, and reduced the Bax/Bcl-2 ratio. Molecular docking results indicated that norwogonin exhibited strong binding abilities with Nrf2 and HO-1.
CONCLUSION Norwogonin alleviates oxidative stress, reduces inflammatory responses, and inhibits apoptosis by activating Nrf2/HO-1 signaling pathway, thereby improving AHH induced lung tissue injury in mice.

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