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湖北中医药大学,武汉 430065
刘鑫,在读硕士,从事治疗失眠相关的中药药理学研究,E-mail:741248163@qq.com
* 张舜波,讲师,从事中药学研究和新药开发,E-mail:693757317@qq.com
收稿日期:2021-02-28,
网络出版日期:2021-06-11,
纸质出版日期:2021-08-20
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刘鑫,游秋云,王平等.酸枣仁汤对老年慢性快动眼睡眠剥夺模型大鼠肝脏线粒体能量代谢及其机制的影响[J].中国实验方剂学杂志,2021,27(16):53-58.
LIU Xin,YOU Qiu-yun,WANG Ping,et al.Effect of Suanzaoren Tang on Energy Metabolism of Liver Mitochondria in Aged Rats with Chronic Rapid Eye Movement Sleep Deprivation and Its Mechanism[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(16):53-58.
刘鑫,游秋云,王平等.酸枣仁汤对老年慢性快动眼睡眠剥夺模型大鼠肝脏线粒体能量代谢及其机制的影响[J].中国实验方剂学杂志,2021,27(16):53-58. DOI: 10.13422/j.cnki.syfjx.20211501.
LIU Xin,YOU Qiu-yun,WANG Ping,et al.Effect of Suanzaoren Tang on Energy Metabolism of Liver Mitochondria in Aged Rats with Chronic Rapid Eye Movement Sleep Deprivation and Its Mechanism[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(16):53-58. DOI: 10.13422/j.cnki.syfjx.20211501.
目的
2
研究酸枣仁汤对老年慢性睡眠剥夺模型大鼠肝脏线粒体能量代谢的影响及作用机制。
方法
2
将50只雄性Wistar大鼠随机分为正常组,模型组、舒乐安定组(0.18 mg·kg
-1
·d
-1
),酸枣仁汤低、高剂量组(6.48,12.96 g·kg
-1
·d
-1
),除正常组外,其余各组均皮下注射
D
-半乳糖,于末次给药后进行多平台水环境睡眠剥夺,造模结束后各组开始灌胃给药,连续给药7 d。采用透射电镜观察肝脏线粒体形态,采用比色法检测大鼠肝脏三磷酸腺苷(ATP)含量的变化,采用比色法检测大鼠肝脏线粒体呼吸链还原型辅酶Ⅰ(复合物Ⅰ),琥珀酸氧化还原酶(复合物Ⅱ),细胞色素C氧化还原酶(复合物Ⅲ),细胞色素C氧化酶(复合物Ⅳ)活性,采用蛋白免疫印迹法(Western blot)检测大鼠肝脏柠檬酸合酶(CS),异柠檬酸脱氢酶(IDH),负责编码ATP合成酶F0亚基的一段核基因(ATP5F1)蛋白的表达水平。
结果
2
与正常组比较,模型组大鼠肝脏内线粒体损伤较严重,线粒体发生肿胀、变形,嵴断裂并且数量有一定的减少,与模型组比较,舒乐安定组、酸枣仁汤高剂量组线粒体损伤减轻,肿胀、变形情况显著减少,嵴断裂减少,其中酸枣仁汤高剂量改善较为明显。与正常组比较,模型组大鼠肝脏ATP含量显著降低(
P
<
0.01),线粒体呼吸链复合物Ⅰ,Ⅱ,Ⅲ,Ⅳ活性均显著降低(
P
<
0.01),IDH,CS,ATP5F1蛋白表达均显著降低(
P
<
0.01);与模型组比较,酸枣仁汤高剂量组肝脏ATP含量显著升高(
P
<
0.01),线粒体呼吸链复合物Ⅰ,Ⅱ,Ⅲ,Ⅳ活性均明显升高(
P
<
0.05,
P
<
0.01),IDH,CS,ATP5F1蛋白表达均显著升高(
P
<
0.01),舒乐安定组肝脏ATP含量明显升高(
P
<
0.05),线粒体呼吸链复合物Ⅰ,Ⅲ活性显著升高(
P
<
0.01),CS,ATP5F1蛋白表达均显著升高(
P
<
0.01),酸枣仁汤低剂量组肝脏线粒体呼吸链复合物Ⅰ,Ⅲ活性明显升高(
P
<
0.05,
P
<
0.01),ATP5F1蛋白表达明显升高(
P
<
0.05)。
结论
2
酸枣仁汤可以改善老年慢性睡眠剥夺引起的肝脏能量代谢异常,其机制可能与上调线粒体电子链酶活性,CS,IDH及ATP5F1蛋白表达有关。
Objective
2
To study the effect of Suanzaoren Tang on energy metabolism of liver mitochondria in aged rats with chronic rapid eye movement (REM) sleep deprivation.
Method
2
Fifty male Wistar rats were randomly divided into the control group, model group, estazolam group (0.18 mg·kg
-1
·d
-1
), and low- (6.48 g·kg
-1
·d
-1
) and high-dose (12.96 g·kg
-1
·d
-1
) Suanzaoren Tang groups. Rats in all groups except for the control group received subcutaneous injection of
D
-galactose and then were deprived of sleep using the multiple platform method after the last administration. Following successful modeling, the rats in each group were treated with intragastric administration of the corresponding drugs for seven consecutive days. The morphology of liver mitochondria was observed under the transmission electron microscope. The content of adenosine triphosphate (ATP) in rat liver was detected by colorimetry, and the activities of ubiquinone oxidoreductase (complex Ⅰ), succinate-ubiquinone oxidoreductase (complex Ⅱ), ubiquinol-cytochrome c oxidoreductase (complex Ⅲ), and cytochrome c oxidase (complex Ⅳ) in mitochondrial respiratory chain of rat liver were measured by colorimetry. The protein expression levels of citrate synthase (CS), isocitrate dehydrogenase (IDH), and ATP synthase, H
+
transporting, mitochondrial F0 complex, subunit b, isoform 1 (ATP5F1) in rat liver were assayed by Western blot.
Result
2
The mitochondrial damage in rat liver of the model group was more serious than that in the control group, manifested as mitochondrial swelling and deformation as well as cristae rupture and reduction. The comparison with the model group revealed that both the positive control and Suanzaoren Tang at the high dose obviously alleviated the mitochondrial swelling and deformation and reduced cristae rupture, with better improvements observed in the high-dose Suanzaoren Tang group. Compared with the control group, the content of ATP, the activities of mitochondrial respiratory chain complexes Ⅰ, Ⅱ, Ⅲ, and Ⅳ, and the protein expression levels of IDH, CS, and ATP5F1 in rat liver of the model group were all significantly decreased (
P<
0.01). Compared with the model group, the content of ATP, the activities of mitochondrial respiratory chain complexes Ⅰ, Ⅱ, Ⅲ, and Ⅳ, and the protein expression levels of IDH, CS, and ATP5F1 in rat liver of the high-dose Suanzaoren Tang group were all significantly increased (
P<
0.05,
P<
0.01). In the positive control group, the content of ATP, the activities of mitochondrial respiratory chain complexes I and Ⅲ, and the protein expression levels of CS and ATP5F1 in rat liver were significantly increased (
P<
0.05,
P<
0.01). The activities of mitochondrial respiratory chain complexes Ⅰ and Ⅲ and the ATP5F1 protein expression in the low-dose Suanzaoren Tang group were significantly elevated (
P<
0.05,
P<
0.01).
Conclusion
2
Suanzaoren Tang alleviates the abnormal liver energy metabolism induced by chronic REM sleep deprivation in the elderly rats, which may be related to its enhancement of mitochondrial electron transport chain enzyme activities and the up-regulation of protein expression levels of CS, IDH and ATP5F1.
马文静 , 冯蓓蕾 , 延梦圆 . 睡眠剥夺与认知障碍研究进展 [J]. 中国神经免疫学和神经病学杂志 , 2020 , 27 ( 5 ): 403 - 406 .
李鹏翔 , 胡爽 , 张榕 , 等 . 睡眠剥夺小鼠肝脏代谢稳态变化的代谢组学分析 [J]. 现代预防医学 , 2020 , 47 ( 4 ): 702 - 706 .
陈娇凤 , 黄剑浩 , 叶颖颖 , 等 . 推针治疗老年慢性失眠26例 [J]. 中国针灸 , 2021 , 41 ( 2 ): 153 - 154 .
黄晓宇 , 谢光璟 , 李浩 , 等 . 天王补心丹加减通过orexin A/OX1R对慢性睡眠剥夺小鼠糖脂代谢的干预作用 [J]. 中国实验方剂学杂志 , 2021 , 27 ( 1 ): 121 - 127 .
高文康 , 舒艳芸 , 叶进 , 等 . 生物节律与肝脏能量代谢 [J]. 世界华人消化杂志 , 2020 , 28 ( 20 ): 1025 - 1035 .
龙清华 , 赵宾宾 , 丁莉 , 等 . 基于PI3K/AKT/GSK-3 β 信号通路探讨酸枣仁汤对APP/PS1双转基因痴呆小鼠海马神经元突触损伤的改善作用 [J]. 中华中医药杂志 , 2020 , 35 ( 5 ): 2546 - 2551 .
刘莹 , 管彤 , 梁昊都 , 等 . 酸枣仁改善睡眠药理作用及其机制研究 [J]. 中医药信息 , 2021 , 38 ( 3 ): 82 - 86 .
张如意 . 酸枣仁汤对基于5HTR调控老年失眠SCN及VLPO系统的作用及机制研究 [D]. 武汉 : 湖北中医药大学 , 2017 .
张如意 , 游秋云 , 王平 , 等 . 酸枣仁汤对慢性睡眠剥夺老年失眠大鼠心肌细胞凋亡及相关蛋白Bcl2、Bax表达的影响 [J]. 中华中医药杂志 , 2017 , 32 ( 4 ): 1691 - 1693 .
赵凡凡 , 周玉枝 , 高丽 , 等 . D -半乳糖致衰老大鼠模型的研究进展 [J]. 药学学报 , 2017 , 52 ( 3 ): 347 - 354 .
徐智 , 吴国明 , 钱桂生 , 等 . 大鼠衰老模型的初步建立 [J]. 第三军医大学学报 , 2003 , 25 ( 4 ): 312 - 315 .
李洁 . 急性运动对大鼠肝脏线粒体呼吸链酶复合物的影响 [J]. 中国运动医学杂志 , 2008 , 27 ( 3 ): 363 - 365 .
翟艺宗 , 黄昌林 , 常祺 , 等 . 脉冲电流经皮刺激肝区对运动性疲劳大鼠肝脏线粒体呼吸链酶复合物活性的影响 [J]. 现代生物医学进展 , 2015 , 15 ( 17 ): 3236 - 3239 .
陈宁 , 宋囡 , 贾连群 , 等 . 化瘀祛痰方对AS家兔肝脏线粒体呼吸链酶复合物的影响 [J]. 中国实验方剂学杂志 , 2018 , 24 ( 7 ): 171 - 176 .
程岩岩 , 刘路 , 贾连群 , 等 . 健脾化痰祛瘀方对AS巴马小型猪心肌细胞线粒体呼吸链复合物和ATP活性的影响 [J]. 辽宁中医杂志 , 2018 , 45 ( 11 ): 2438 - 2441 .
何清湖 , 宾东华 , 周青 , 等 . 知柏地黄汤对解脲脲原体感染模型大鼠精子线粒体呼吸链复合物的影响 [J]. 中华男科学杂志 , 2016 , 22 ( 11 ): 1005 - 1010 .
MUSTER B , KOHL W , WITTIG I , et al . Respiratory chain complexes in dynamic mitochondria display a patchy distribution in life cells [J]. PLoS One , 2010 , 5 ( 7 ): 11910 - 11919 .
杨旭 , 王琦越 , 刘渊 , 等 . 从“脾主化”角度探讨湿阻中焦证大鼠线粒体能量代谢路径的影响及平胃散的干预作用 [J]. 时珍国医国药 , 2019 , 30 ( 6 ): 1305 - 1308 .
姚凤云 , 彭淑红 , 左铮云 , 等 . 桂枝甘草汤对低温环境大鼠心肌PGC1α、CS、α-KGDHC含量的影响 [J]. 时珍国医国药 , 2015 , 26 ( 2 ): 263 - 264 .
屈小虎 , 黄玲 , 陈慧 , 等 . 脾气虚证和脾不统血证模型大鼠脾主肌肉实质的探讨 [J]. 中国中西医结合杂志 , 2019 , 39 ( 2 ): 217 - 222 .
王志丹 , 宋囡 , 吕美君 , 等 . 益气健脾化痰祛瘀方对脾虚痰浊动脉粥样硬化巴马小型猪肝脏线粒体能量代谢的影响 [J]. 中华中医药杂志 , 2017 , 32 ( 9 ): 4107 - 4110 .
RODGERS A J , WILKENS S , AGGELER R , et al . The subunitδsub-unit b domain of the Escherichia coli F1 FoATPase.The b subunits interact with F1as a dimer and through theδsubunit [J]. J Biol Chem , 1997 , doi: 10.1074/jbc.272.49.31058 http://dx.doi.org/10.1074/jbc.272.49.31058 .
张吉斌 , 刘月平 , 方美英 . 线粒体ATP合酶基因组成及生化机制研究进展 [J]. 中国畜牧杂志 , 2010 , 46 ( 7 ): 64 - 68 .
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