中医药学报

红大戟醇提物对缺氧诱导的小鼠肺上皮细胞损伤的保护作用及机制

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中医药学报 | 实验研究 2026,54(3): 24-33

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中医药学报 |实验研究 2026 , 54 (3) : 24 -33

红大戟醇提物对缺氧诱导的小鼠肺上皮细胞损伤的保护作用及机制

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杨秀玲¹, 李昊¹, 龙云¹, 杨真琴¹, 申利¹, 邱斌¹, 李学芳¹, 张范¹

作者信息

通讯作者:

* 张范（1982-），男，博士，副研究员，硕士生导师，主要从事分子药理学研究。

作者简介:

杨秀玲（2000-），女，硕士研究生，专业：药物分析学。

Protective Effect and Mechanism of Knoxia Roxburghii Ethanol Extract on Hypoxia - Induced Lung Epithelial Cell Injury in Mice

YANG Xiuling¹, LI Hao¹, LONG Yun¹, YANG Zhenqin¹, SHEN Li¹, QIU Bin¹, LI Xuefang¹, ZHANG Fan¹

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doi: 10.19664/j.cnki.1002-2392.260050

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摘要

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目的： 探讨红大戟醇提物对缺氧诱导的小鼠肺上皮细胞TC - 1损伤的保护作用及机制。方法： 利用液相色谱 -质谱联用仪（LC - MS）分析红大戟醇提物中的化合物。通过三气培养箱（37 ℃、5%CO₂、94% N₂、1% O₂）建立细胞缺氧模型。将TC - 1细胞分为常氧组（Normal组）、缺氧组（Hypoxia组）、缺氧+红大戟醇提物低剂量组（Hypoxia + KRL组，90 μg/mL）、缺氧+红大戟醇提物高剂量组（Hypoxia +KRH组，180 μg/mL）。CCK - 8法测定细胞活力；倒置显微镜观察细胞形态变化并统计细胞个数；检测细胞培养上清液中乳酸脱氢酶（LDH）的泄露量及细胞中丙二醛（MDA）、超氧化物歧化酶（SOD）、肿瘤坏死因子 - α（TNF - α）、白细胞介素 - 6（IL - 6）及ATP含量；荧光显微镜观察细胞中活性氧（ROS）荧光强度；RT - qPCR法检测缺氧诱导因子- 1α（HIF - 1α）、内皮一氧化氮合酶（eNOS）、诱导型一氧化氮合酶（iNOS）的mRNA表达量；Western blot法检测细胞中HIF - 1α、eNOS、iNOS的蛋白表达水平。结果： 在红大戟醇提物中找到22个化合物。TC - 1细胞经过24 h的缺氧处理后，细胞肿大变圆、死细胞增多，ROS（P < 0.01）、LDH（P < 0.01）、MDA（P < 0.01）、TNF - α（P < 0.01）及IL - 6（P < 0.01）含量显著升高，SOD（P < 0.01）和ATP（P < 0.01）含量则显著降低，HIF - 1α（P < 0.01）与iNOS（P < 0.01）的mRNA及蛋白表达水平显著升高，eNOS（P < 0.01）的表达水平则降低。在红大戟醇提物处理后细胞数量显著增加，细胞形态趋于Normal组，ROS（P < 0.01）、MDA（P < 0.05）、TNF - α（P < 0.01）及IL - 6（P < 0.01）含量降低；HIF - 1α（P < 0.05）、iNOS（P < 0.05）的mRNA及蛋白表达水平降低；SOD（P <0.01）、ATP（P < 0.01）、eNOS（P < 0.05）则显著升高。结论： 红大戟醇提物在TC - 1细胞受到缺氧刺激时能够下调HIF - 1α的表达，改善细胞能量代谢紊乱、氧化应激反应和炎症反应来发挥抗缺氧作用。

关键词

红大戟 / 缺氧损伤 / 氧化应激 / HIF - 1α/eNOS

Abstract

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Objective: To investigate the protective effect and mechanism of KR ethanol extract on hypoxia - induced TC - 1 injury in mouse lung epithelial cells.Methods: LC - MS was used to analyze the compounds in the alcohol extract of KR. The cell hypoxia model was established by three - gas incubator (37 ℃, 5% CO₂, 94% N₂, 1% O₂). TC - 1 cells were divided into normal group (Normal), hypoxia group (Hypoxia), hypoxia + KR alcohol extract low dose group (Hypoxia + KRL, 90 μg/mL), hypoxia + KR alcohol extract high dose group (Hypoxia + KRH, 180 μg/mL). Cell viability was measured by CCK - 8 assay. The morphological changes of cells were observed by inverted microscope and the number of cells was counted. The leakage of lactate dehydrogenase (LDH) in the supernatant of cell culture medium and the contents of malondialdehyde (MDA), superoxide dismutase (SOD), tumor necrosis factor - α (TNF - α), interleukin -6 (IL - 6) and ATP in cells were detected. The fluorescence intensity of reactive oxygen species (ROS) in cells was observed by fluorescence microscope. The mRNA expression levels of hypoxia - inducible factor - 1α (HIF - 1α), endothelial nitric oxide synthase (eNOS) and inducible nitric oxide synthase (iNOS) were detected by RT - qPCR. The protein expression levels of HIF - 1α, eNOS and iNOS in cells were detected by Western blot.Results: Twenty - two compounds were found in the KR ethanol extract. After 24 h of hypoxia treatment, the cells of TC - 1 cells became swollen and round, and the contents of ROS, LDH, MDA, TNF - α and IL - 6 were significantly increased, while the contents of SOD and ATP were significantly decreased. The mRNA and protein expression levels of HIF - 1α and iNOS were significantly increased, while the expression level of eNOS was decreased. After treatment with KR ethanol extract, the number of cells increased significantly, the cell morphology tended to be Normal group, and the contents of ROS, LDH, MDA, TNF - α and IL - 6 decreased. The mRNA and protein expression levels of HIF - 1α and iNOS were decreased. SOD, ATP and eNOS were significantly increased.Conclusion: KR ethanol extract can down - regulate the expression of HIF - 1α when TC - 1 cells are stimulated by hypoxia, and improve cell energy metabolism disorder, oxidative stress response and inflammatory response to exert anti - hypoxia effect.

Key words

Knoxia roxburghii / Hypoxia injury / Oxidative stress / HIF - 1α/eNOS

引用本文

杨秀玲, 李昊, 龙云, 杨真琴, 申利, 邱斌, 李学芳, 张范. 红大戟醇提物对缺氧诱导的小鼠肺上皮细胞损伤的保护作用及机制. 中医药学报, 2026 , 54 (3) : 24 -33 . DOI: 10.19664/j.cnki.1002-2392.260050

YANG Xiuling, LI Hao, LONG Yun, YANG Zhenqin, SHEN Li, QIU Bin, LI Xuefang, ZHANG Fan. Protective Effect and Mechanism of Knoxia Roxburghii Ethanol Extract on Hypoxia - Induced Lung Epithelial Cell Injury in Mice[J]. Acta Chinese Medicine and Pharmacology, 2026 , 54 (3) : 24 -33 . DOI: 10.19664/j.cnki.1002-2392.260050

基金

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云南省基础研究计划项目 (202101AZ070001-006); 云南省生物医药重大专项 (202302AA310014)

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2026年第54卷第3期

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doi: 10.19664/j.cnki.1002-2392.260050

接收时间：2025-01-22
首发时间：2026-09-07

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收稿日期：2025-01-22

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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