中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (20): 3212-3219.doi: 10.12307/2022.623

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

铁皮石斛黄酮干预力竭运动模型大鼠肝脏氧化应激及自噬水平的变化

王思涵1，陈俊吉1，莫伟彬2，3   

  1. 1广西师范大学漓江学院，广西壮族自治区桂林市  541006；2广西师范大学体育与健康学院，广西壮族自治区桂林市  541004；3省部共建药用资源化学与药物分子工程国家重点实验室，广西壮族自治区桂林市  541004
  • 收稿日期:2021-08-23 接受日期:2021-10-11 出版日期:2022-07-18 发布日期:2022-01-19
  • 通讯作者: 莫伟彬，博士，副教授，广西师范大学体育与健康学院，广西壮族自治区桂林市 541004；省部共建药用资源化学与药物分子工程国家重点实验室，广西壮族自治区桂林市 541004
  • 作者简介:王思涵，女，1978年生，汉族，广西壮族自治区桂林市人，2003年上海体育学院毕业，讲师，主要从事运动生理学与训练研究。
  • 基金资助:
    广西教育厅科研项目(2021KY1596)，项目负责人：陈俊吉；广西师范大学教育发展基金会“教师成长基金”项目(EDF2016005)，项目负责人：莫伟彬

Effects of Dendrobium officinale flavonoid on oxidative stress and autophagy in the liver of an exhaustive exercise rat model

Wang Sihan1, Chen Junji1, Mo Weibin2, 3   

  1. 1Lijiang College, Guangxi Normal University, Guilin 541006, Guangxi Zhuang Autonomous Region, China; 2College of Physical Education and Health, Guangxi Normal University, Guilin 541004, Guangxi Zhuang Autonomous Region, China; 3State Key Laboratory of Medicinal Resources Chemistry and Molecular Engineering, Guilin 541004, Guangxi Zhuang Autonomous Region, China
  • Received:2021-08-23 Accepted:2021-10-11 Online:2022-07-18 Published:2022-01-19
  • Contact: Mo Weibin, MD, Associate professor, College of Physical Education and Health, Guangxi Normal University, Guilin 541004, Guangxi Zhuang Autonomous Region, China; State Key Laboratory of Medicinal Resources Chemistry and Molecular Engineering, Guilin 541004, Guangxi Zhuang Autonomous Region, China
  • About author:Wang Sihan, Lecturer, Lijiang College, Guangxi Normal University, Guilin 541006, Guangxi Zhuang Autonomous Region, China
  • Supported by:
    the Research Project of Education Department in Guangxi Zhuang Autonomous Region, No. 2021KY1596 (to CJJ); Teacher Development Fund of Education Development Foundation of Guangxi Normal University, No. EDF2016005 (to MWB)

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摘要：

文题释义：
氧化应激：是指体内氧化与抗氧化作用失衡的一种状态，倾向于氧化，导致中性粒细胞炎性浸润，蛋白酶分泌增加，产生大量氧化中间产物。氧化应激是由自由基在体内产生的一种负面作用，并被认为是导致衰老和疾病的一个重要因素。
自噬：是指机体细胞受到内外环境不利因素刺激时，机体为维持自身内环境稳态的消化代谢过程，将胞质中的细胞器、蛋白和大分子等物质重建、分解或更新后再利用；从细胞成分运输至溶酶体角度划分，可将自噬分为大自噬(巨自噬)、伴侣蛋白介导的自噬和微自噬。
自噬体：在自噬形成的过程中，随着自噬泡的进一步改变，细胞器进入到囊泡膜内，形成新的囊泡核，称为自噬体。

背景：适当的运动能够提高机体的抗氧化能力，抑制和延缓氧化应激的发生，激活机体内细胞自噬。但长时间的耐力训练后至力竭运动大鼠肝脏组织氧化应激及自噬机制十分复杂，铁皮石斛黄酮具有抗氧化、调节免疫和抗炎镇痛等作用，运动前补充铁皮石斛黄酮是否能达到保护肝脏组织的目的目前尚不明确。
目的：探究铁皮石斛黄酮对力竭性运动大鼠肝脏组织氧化应激及自噬水平的影响。
方法：将60只10周龄雄性SD大鼠随机分为安静对照组、运动对照组、铁皮石斛黄酮低、中、高剂量组。除安静对照组外，其余大鼠进行为期5 d的适应性跑台训练，之后进行为期6周的正式运动训练，于6周末即最后一次训练达到力竭。铁皮石斛黄酮低、中、高剂量组大鼠分别在运动前半小时灌胃1.5，3.0，4.5 g/(kg•d)的铁皮石斛黄酮，灌胃体积为5 mL/kg，1次/d，直到实验结束。苏木精-伊红染色观察大鼠肝脏组织病理学变化，检测大鼠血清丙氨酸氨基转移酶、天冬氨酸氨基转移酶；丙二醛、过氧化氢酶、超氧化物歧化酶、谷胱甘肽过氧化物酶表达水平和总抗氧化能力，RT-qPCR和Western blotting测试肝脏组织自噬相关指标的变化。 
结果与结论：① 苏木精-伊红染色后运动对照组大鼠肝细胞索排列紊乱，肝窦肿胀、汇管区有炎性细胞浸润，胞质内有大量的脂滴；铁皮石斛黄酮低剂量组大鼠肝组织脂滴和炎症细胞浸润明显减少；中、高剂量组大鼠肝组织与安静对照组比较无明显变化；②与安静对照组比较，运动对照组大鼠血清丙氨酸氨基转移酶、天冬氨酸氨基转移酶、丙二醛表达水平及总抗氧化能力均明显升高(P < 0.01)，血清过氧化氢酶、超氧化物歧化酶及谷胱甘肽过氧化物酶活性均明显降低(P < 0.01)；与运动对照组比较，铁皮石斛黄酮低、中、高剂量组丙氨酸氨基转移酶及天冬氨酸氨基转移酶活性均出现降低(P < 0.01)，血清过氧化氢酶、超氧化物歧化酶及谷胱甘肽过氧化物酶活性均明显升高(P < 0.01)；铁皮石斛黄酮中、高剂量组大鼠血清丙二醛水平及总抗氧化能力均出现降低(P < 0.05)；③与安静对照组比较，运动对照组大鼠肝组织Beclin1、Atg7、LC3-Ⅱ、p62及 Rab7 mRNA及蛋白表达水平均明显升高(P < 0.01)；与运动对照组比较，铁皮石斛黄酮中、高剂量组大鼠肝组织Beclin1、Atg7、LC3-Ⅱ、p62及 Rab7 mRNA及蛋白表达水平均出现下降趋势(P < 0.05或P < 0.01)；④提示长期的耐力运动至力竭运动会引起大鼠肝组织氧化应激、肝组织受损，铁皮石斛黄酮通过调节肝脏氧化代谢和调控大鼠肝脏组织自噬水平维持肝脏功能的稳定，从而达到保护肝脏的目的。
缩略语：丙氨酸氨基转移酶：alanine aminotransferase，ALT；天冬氨酸氨基转移酶：aspartate aminotransferase，AST

https://orcid.org/0000-0003-1530-5803 (王思涵) 

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 铁皮石斛黄酮, 力竭运动, 肝脏, 氧化应激, 自噬, 大鼠

Abstract: BACKGROUND: Proper exercise can improve antioxidant capacity, inhibit and delay oxidative stress, and activate autophagy in the body. However, the mechanism of oxidative stress and autophagy in the liver tissue of rats after exhaustive exercise due to a long-term endurance training is still very complicated. Dendrobium officinale flavonoids have antioxidant capacity, immune regulation, anti-inflammatory and analgesic effects. It is still unclear whether Dendrobium officinale flavonoids can protect liver tissue before exercise. 
OBJECTIVE: To investigate the effect of Dendrobium officinale flavonoids on oxidative stress and autophagy in the liver tissue of exhaustive exercise rats.
METHODS: Sixty male Sprague-Dawley rats aged 10 weeks old were randomly divided into a quiet control group, an exercise control group, and low-, medium-, and high-dose Dendrobium officinale flavonoid groups. Except for the quiet control group, the rest rats underwent 5-day adaptive treadmill training, followed by 6-week formal exercise training. Exhaustion was achieved in the last training at the end of 6 weeks. Rats in the low-, medium- and high-dose Dendrobium officinale flavonoids groups were intragastrically administrated with 1.5, 3.0, and 4.5 g/(kg•d) Dendrobium officinale flavonoids half an hour before exercise, respectively. The intragastric volume was 5 mL/kg, once a day, until the end of the experiment. Hematoxylin-eosin staining was used to observe the pathological changes of rat liver tissue, and to detect rat serum alanine aminotransferase and aspartate aminotransferase, malondialdehyde, catalase, superoxide dismutase, and glutathione peroxidase levels, and total antioxidant capacity. Changes in autophagy-related indexes in rat liver tissue were detected by RT-qPCR and western blot assay. 
RESULTS AND CONCLUSION: Hematoxylin-eosin staining results revealed that in the exercise control group, hepatocytes were arranged in a disorderly manner, the hepatic sinusoids were swollen, the portal area was infiltrated with inflammatory cells, and there were a large number of lipid droplets in the cytoplasm. The lipid droplets and inflammatory cell infiltration in the liver tissue of rats were significantly reduced in the low-dose Dendrobium officinale flavonoid group, while there were no significant changes in the liver tissue of the rats in the medium- and high-dose Dendrobium officinale flavonoid groups compared with the quiet group. Compared with the quiet control group, the expression levels of serum alanine aminotransferase, aspartate aminotransferase, malondialdehyde and total antioxidant capacity were significantly increased (P < 0.01), while the serum activities of catalase, superoxide dismutase and glutathione peroxidase were significantly decreased in the exercise control group (P < 0.01). Compared with the exercise control group, the activities of alanine aminotransferase and aspartate aminotransferase were significantly decreased (P < 0.01), and the activities of serum catalase, superoxide dismutase and glutathione peroxidase were significantly increased in the low-, medium- and high-dose Dendrobium officinale flavonoid groups (P < 0.01). Moreover, the serum level of malonaldehyde and total antioxidant capacity were decreased in the medium- and high-dose Dendrobium officinale flavonoid groups (P < 0.05). Compared with the quiet control group, the mRNA and protein expression levels of Beclin1, Atg7, LC3-II, p62 and Rab7 in the liver tissue of rats were significantly increased in the exercise control group (P < 0.01). Compared with the exercise control group, the mRNA and protein expression levels of Beclin1, Atg7, LC3 - II, p62 and Rab7 in the liver tissues of rats showed a downward trend in the medium- and high-dose Dendrobium officinale flavonoid groups (P < 0.05 or P < 0.01). To conclude, long-term endurance exercise until exhaustive exercise can induce oxidative stress and damage to liver tissue in rats. Dendrobium officinale flavonoids can maintain the stability of liver function by regulating liver oxidative metabolism and autophagy in rat liver tissue. Thereby, liver protection is achieved.

Key words: Dendrobium officinale flavonoid, exhaustive exercise, liver, oxidative stress, autophagy, rat

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