辅酶Q10改善LPS诱导小鼠急性肺损伤的效应分析

doi:10.11843/j.issn.0366-6964.2023.04.036

摘要/Abstract

摘要： 本研究旨在探究辅酶Q10（coenzyme Q10，CoQ10）对脂多糖（lipopolysaccharide，LPS）致小鼠急性肺损伤的保护作用及机制。选取8周龄的健康雄性C57BL/6小鼠72只，随机均分为6组：空白对照组（CON组，100 μL玉米油）、CoQ10干预组（CHQ组，50 mg·kg^-1 CoQ10）、模型组（LPS组，100 μL玉米油+LPS）、低剂量CoQ10处理组（LDQ组，2 mg·kg^-1 CoQ10+LPS）、中剂量CoQ10处理组（LMQ组，10 mg·kg^-1 CoQ10+LPS）和高剂量CoQ10处理组（LHQ组，50 mg·kg^-1 CoQ10+LPS）。通过灌胃的方式连续给予小鼠不同剂量的CoQ10或玉米油14 d，然后鼻内滴入50 μL 1 mg·mL^-1 LPS溶液建立小鼠急性肺损伤模型。24 h后，处死小鼠，摘取左肺称重并烘干，计算肺组织湿干质量比（W/D）；检测肺泡灌洗液（BALF）蛋白浓度、肺组织丙二醛（MDA）含量、活性氧（ROS）浓度、总超氧化物歧化酶（SOD）活性和还原型谷胱甘肽（GSH）水平；苏木素伊红（HE）染色观察肺组织病理变化；TUNEL染色法检测肺组织细胞凋亡率；RT-PCR技术检测线粒体细胞凋亡途径相关基因表达；Western blot法检测肺组织MKP-1/Nrf2通路相关蛋白表达。结果表明，与CON组相比，LPS组小鼠肺组织W/D和BALF蛋白浓度极显著升高（P<0.01）；肺组织出血、肺泡壁增厚、炎性细胞浸润；MDA和ROS含量极显著升高（P<0.01），SOD活性和GSH含量极显著降低（P<0.01）；TUNEL凋亡率和线粒体凋亡途径相关基因表达均极显著升高（P<0.01）；MKP-1和Nrf2蛋白表达极显著降低（P<0.01）。低剂量、中剂量和高剂量CoQ10均显著或极显著地逆转了LPS诱导的上述指标的变化（P<0.05，P<0.01），且高剂量CoQ10效果更显著（P<0.01）。此外，CHQ组与CON组相比，所有指标均无显著性差异（P>0.05）。结果提示，CoQ10可通过激活MKP-1/Nrf2信号通路，增强小鼠抗氧化应激能力，减少ROS产生，抑制线粒体细胞凋亡，从而改善LPS诱导的小鼠急性肺损伤。

关键词: 辅酶Q10, 急性肺损伤, 氧化应激, 线粒体细胞凋亡, MKP-1/Nrf2信号通路

Abstract: The purpose of this study was to investigate the protective effect and mechanism of coenzyme Q10 (CoQ10) on lipopolysaccharide (LPS)-induced acute lung injury in mice. Seventy-two healthy male C57BL/6 mice aged 8 weeks were selected and randomly divided into 6 groups:control group (CON group, 100 μL corn oil), CoQ10 intervention group (CHQ group, 50 mg·kg^-1 CoQ10), model group (LPS group, 100 μL corn oil + LPS), low-dose CoQ10 treatment group (LDQ group, 2 mg·kg^-1 CoQ10 + LPS), medium-dose CoQ10 treatment group (LMQ group, 10 mg·kg^-1 CoQ10 + LPS) and high-dose CoQ10 treatment group (LHQ group, 50 mg·kg^-1 CoQ10+LPS). The mice were continuously given different doses of CoQ10 or corn oil by gavage for 14 days, and intranasally instilled 50 μL 1 mg·mL^-1 LPS to establish an acute lung injury model. After 24 h, the mice were sacrificed, and the left lung was harvested, weighed and dried to calculate the wet-dry weight ratio (W/D) of lung tissue. The protein concentration of alveolar lavage fluid (BALF), malondialdehyde (MDA) content, reactive oxygen species (ROS) concentration, total superoxide dismutase (SOD) activity and reduced glutathione (GSH) level in lung tissue were detected. Hematoxylin and eosin (HE) staining was used to observe the pathological changes of lung tissues. TUNEL staining was used to detect the apoptosis rate of lung tissue. RT-PCR technology was used to detect the expression of mitochondrial apoptosis pathway-related genes. Western blot was used to detect the expression of MKP-1/Nrf2 pathway-related proteins in lung tissue. The results showed that compared with CON group, the W/D value and BALF protein concentration in the LPS group were significantly increased (P<0.01); lung tissue hemorrhage, alveolar wall thickening, inflammatory cell infiltration; MDA and ROS contents were significantly increased (P<0.01), SOD activity and GSH content were significantly decreased (P<0.01); TUNEL apoptosis rate and mitochondrial apoptosis pathway related gene expression were significantly increased (P<0.01); MKP-1 and Nrf2 protein expression were significantly decreased (P<0.01). Low-dose, medium-dose and high-dose CoQ10 significantly or extremely significantly reversed the changes of the above indexes induced by LPS (P<0.05, P<0.01), and high-dose CoQ10 was more effective (P<0.01). In addition, there was no significant difference in all indicators between the CHQ group and the CON group (P>0.05). The results suggest that CoQ10 can improve LPS-induced acute lung injury in mice by activating MKP-1/Nrf2 signaling pathway, enhancing anti-oxidative stress ability, reducing ROS production and inhibiting mitochondrial apoptosis.

Key words: coenzyme Q10, acute lung injury, oxidative stress, mitochondrial apoptosis, MKP-1/Nrf2 signaling pathway

中图分类号:

S857.11

陈永平, 寇玉红, 焦文静, 侯晓昱, 范宏刚. 辅酶Q10改善LPS诱导小鼠急性肺损伤的效应分析[J]. 畜牧兽医学报, 2023, 54(4): 1730-1741.

CHEN Yongping, KOU Yuhong, JIAO Wenjing, HOU Xiaoyu, FAN Honggang. Effect of Coenzyme Q10 on LPS-induced Acute Lung Injury in Mice[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1730-1741.

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