张兆强, 解瑞宁, 韩贵芝, 邵波, 林立, 张春芝. 染尘大鼠肺组织中SDHA mRNA水平及线粒体ATP代谢的变化[J]. 环境与职业医学, 2019, 36(10): 955-959. DOI: 10.13213/j.cnki.jeom.2019.19146
引用本文: 张兆强, 解瑞宁, 韩贵芝, 邵波, 林立, 张春芝. 染尘大鼠肺组织中SDHA mRNA水平及线粒体ATP代谢的变化[J]. 环境与职业医学, 2019, 36(10): 955-959. DOI: 10.13213/j.cnki.jeom.2019.19146
ZHANG Zhao-qiang, XIE Rui-ning, HAN Gui-zhi, SHAO Bo, LIN Li, ZHANG Chun-zhi. Changes of SDHA mRNA and mitochondrial ATP metabolism in lung of dust-exposed rats[J]. Journal of Environmental and Occupational Medicine, 2019, 36(10): 955-959. DOI: 10.13213/j.cnki.jeom.2019.19146
Citation: ZHANG Zhao-qiang, XIE Rui-ning, HAN Gui-zhi, SHAO Bo, LIN Li, ZHANG Chun-zhi. Changes of SDHA mRNA and mitochondrial ATP metabolism in lung of dust-exposed rats[J]. Journal of Environmental and Occupational Medicine, 2019, 36(10): 955-959. DOI: 10.13213/j.cnki.jeom.2019.19146

染尘大鼠肺组织中SDHA mRNA水平及线粒体ATP代谢的变化

Changes of SDHA mRNA and mitochondrial ATP metabolism in lung of dust-exposed rats

  • 摘要: 背景 职业性矽肺是严重威胁相关从业者生命和健康的呼吸系统疾病,具有渐进性和不可逆性的特点,目前其发生机制及发病进程尚未彻底阐明。

    目的 探讨染尘Wistar大鼠肺组织中琥珀酸脱氢酶A亚基(SDHA)mRNA水平及线粒体三磷酸腺苷(ATP)代谢的变化。

    方法 将40只Wistar大鼠随机分为对照组和3个实验组,每组10只。实验组大鼠一次性经气管向肺内注入1 mL SiO2(100 mg/mL)悬浊液,分别于矽尘染尘后30、60、120 d处死。对照组注入等量生理盐水,120 d后处死。取肺组织,行苏木精-伊红染色法(HE)染色观察其大体病理变化,电镜观察其显微病理变化;实时荧光定量PCR检测SDHA mRNA的相对含量;提取肺组织线粒体,检测其ATP含量及Ca2+-Mg2+-ATP酶活性。

    结果 HE染色结果显示,染尘时间越长,肺纤维化越明显。电镜显示,与对照组比,实验组大鼠肺组织的线粒体出现肿胀及破坏。对照组及3个实验组肺组织SDHA mRNA相对含量分别为1.08±0.34、4.17±0.38、1.96±0.43、0.44±0.40,与对照组相比,30 d组表达量升高,120 d组表达量降低(P < 0.05)。肺组织线粒体ATP含量分别为(402.20±85.18)、(700.35±98.60)、(350.10±92.57)、(200.51±88.49)μmol/g(以每g蛋白计)。线粒体中Ca2+-Mg2+-ATP酶活性分别为(3.96±2.51)×10-2、(9.58±3.83)×10-2、(4.89±4.72)×10-2、(2.51±3.97)×10-2 U/mg(以每g蛋白计)。ATP及Ca2+-Mg2+-ATP酶活性规律一致,均为30 d组高于对照组,120组低于对照组(P < 0.05)。

    结论 矽尘吸入可导致大鼠肺组织SDHA mRNA水平于早期升高至顶峰,随后逐渐降低至正常水平之下,肺组织ATP含量和Ca2+-Mg2+-ATP酶活性发生相应改变。

     

    Abstract: Background Occupational silicosis is a respiratory disease that seriously threatens the life and health of related occupational practitioners. It is progressive and irreversible, and its mechanism and progression have not been fully elucidated.

    Objective This experiment investigates the expression of succinate dehydrogenase (SDHA) mRNA in the lung of dust-exposed Wistar rats and changes of mitochondrial adenosine triphosphate (ATP) metabolism.

    Methods Forty Wistar rats were randomly divided into a control group and three experimental groups, with 10 rats in each group. The rats in the experimental groups were exposed to 1 mL SiO2 (100 mg/mL) suspension by intratracheal instillation for only once, and were neutralized after 30, 60, and 120 days, respectively. The rats in the control group were treated with normal saline, and were neutralized 120 days later. Lung tissues were harvested to observe pathological changes using hematoxylin and eosin (HE) staining and electron microscopy. The relative expression of SDHA mRNA was measured by quantitative real-time PCR. The ATP content and Ca2+-Mg2+-ATPase activity of lung mitochondria were detected.

    Results The pathological images of lung tissues stained with HE showed that pulmonary fibrosis was more obvious over time in the rats of the experimental groups. The pathological images of electron microscopy showed that the mitochondria in lung tissues of the experimental groups were swollen and destroyed compared with the control group. The relative expression levels of SDHA mRNA in the control group and the 30d, 60d, and 120d experimental groups were 1.08±0.34, 4.17±0.38, 1.96±0.43, and 0.44±0.40, respectively. Compared with the control group, the expression of SDHA mRNA in the 30d group was increased and that in the 120d group was reduced (P < 0.05). The contents of mitochondrial ATP in lung were:control group (402.20±85.18)μmol/g (normalized by protein in per g, thereafter), 30d group (700.35±98.60)μmol/g, 60d group (350.10±92.57)μmol/g, and 120d group (200.51±88.49)μmol/g, respectively. The activities of Ca2+-Mg2+-ATPase of mitochondria were:control group (3.96±2.51)×10-2 U/mg (normalized by protein in per g, thereafter), 30 d group (9.58±3.83)×10-2 U/mg, 60 d group (4.89±4.72)×10-2 U/mg, and 120 d group (2.51±3.97)×10-2 U/mg, respectively. The content of ATP and the activity of Ca2+-Mg2+-ATPase changed in the same way, which were higher in the 30 d group and lower in the 120 d group than those in the control group (P < 0.05).

    Conclusion Dust inhalation could cause the expression of SDHA mRNA in rat lung tissues to peak first in an early stage, then gradually fall below normal level, and corresponding changes in ATP content and Ca2+-Mg2+-ATPase activity.

     

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