犬尿氨酸/犬尿酸通路在原发性胆汁性胆管炎伴疲劳小鼠肌肉中的作用及机制研究

The role and mechanism of the kynurenine/kynuric acid pathway in the muscles of mice with primary biliary cholangitis and fatigue

目的探究犬尿氨酸(kynurenine,KYN)/犬尿酸(kynuric acid,KYNA)通路在原发性胆汁性胆管炎伴疲劳小鼠肌肉中的作用及机制。方法随机将12只小鼠分成对照组、DDC组、DDC+抑郁组、DDC+抑郁+氯米帕明组,每组3只,对照组常规饲料喂养,DDC+抑郁组和DDC+抑郁+氯米帕明组采用慢性不可预见性温和应激方法复制小鼠抑郁模型;2周后DDC组、DDC+抑郁组、DDC+抑郁+氯米帕明组同时开始连续喂养含有0.1%的DDC饲料建立胆汁淤积模型,同时DDC+抑郁+氯米帕明组腹腔注射盐酸氯米帕明(7.5 mg·kg^(-1)·d^(-1))连续14 d,其余组小鼠经腹腔注射生理盐水。观察并记录小鼠体质量变化。进行负重游泳实验,记录各组小鼠的负重游泳时间。采用强迫游泳实验和悬尾实验评价小鼠抑郁效果。应用ELISA检测各组小鼠血清和肌肉中KYN、KYNA含量的变化;取肝脏和肌肉进行HE染色,观察肝脏和肌肉组织病理学变化;采用ROS染色分析肌肉氧化损伤的程度。结果与对照组相比,DDC组、DDC+抑郁组和DDC+抑郁+氯米帕明组小鼠表现出明显的疲劳和抑郁样行为(P<0.05);对比DDC组,DDC+抑郁组的疲劳和抑郁样行为加重(P<0.05);DDC+抑郁+氯米帕明组的疲劳和抑郁较DDC+抑郁组减轻(P<0.05)。小鼠肝组织和肌组织HE染色结果显示,相比对照组,DDC组、DDC+抑郁组和DDC+抑郁+氯米帕明组表现出不同程度病理损伤;DDC+抑郁+氯米帕明组小鼠的肝和肌组织病理损伤较DDC+抑郁组减轻。与对照组相比,DDC组、DDC+抑郁组和DDC+抑郁+氯米帕明组小鼠肌肉中ROS含量表现出不同程度升高;与DDC+抑郁组相比,DDC+抑郁+氯米帕明组小鼠肌肉中ROS含量显著下降。相比对照组,DDC组、DDC+抑郁组和DDC+抑郁+氯米帕明组小鼠血清和肌肉中KYN水平显著升高(P<0.05),KYNA水平显著下降(P<0.05);DDC+抑郁组小鼠血清和肌肉中KYN浓度比DDC组明显升高(P<0.05);DDC+抑郁+氯米帕明组小鼠血清和肌肉中KYN浓度比DDC+抑郁组明显降低(P<0.05)。结论KYN/KYNA通路在原发性胆汁性胆管炎伴疲劳小鼠中对外周疲劳有调控作用,抗抑郁治疗能改善肌肉疲劳。

Objective To investigate the role and mechanism of the kynurenine/kynuric acid(KYN/KYNA)pathway in the muscles of mice with primary biliary cholangitis and fatigue.[WTHZ]Methods Twelve mice were randomly divided into control group,DDC group,DDC+depression group,DDC+depression+Clomipramine group,with 3 mice in each group.The control group was fed with conventional feed,while the DDC+depression group and DDC+depression+Clomipramine group were subjected to chronic unpredictable mild stress to replicate mice depression models.Two weeks later,the DDC group,DDC+depression group and DDC+depression+Clomipramine group began to continuously feed with feed containing 0.1%of DDC to establish a bile stasis model.At the same time,the DDC+depression+Clomipramine group was intraperitoneally injected with Clomipramine hydrochloride(7.5 mg·kg^(-1)·d^(-1))for 14 consecutive days,while the other groups of mice were intraperitoneally injected with physiological saline.Observe and record changes in mice body mass.Conduct a weight-bearing swimming test and record the weight-bearing swimming time of each group of mice.Evaluate the depression effect in mice using forced swimming and tail suspension test.ELISA was used to detect the changes in serum and muscle levels of KYN and KYNA in each group of mice.Take the liver and muscles for HE staining,and observe the pathological changes in the liver and muscle tissues.Use ROS staining to analyze the degree of muscle oxidative damage.[WTHZ]Results Compared with the control group,the DDC group,DDC+depression group and DDC+depression+Clomipramine group mice showed significant fatigue and depressive like behavior(P<0.05).Compared with the DDC group,the DDC+depression group showed increased fatigue and depressive like behavior(P<0.05).The fatigue and depression in the DDC+depression+Clomipramine group were reduced compared with the DDC+depression group(P<0.05).The HE staining results of mice liver and muscle tissues showed that compared to the control group,the DDC group,DDC+depression group and DDC+depression+Clomipramine group exhibited varying degrees of pathological damage.The pathological damage of liver and muscle tissues in the DDC+depression+Clomipramine group mice was reduced compared to the DDC+depression group.Compared with the control group,the ROS content in the muscles of mice in the DDC group,DDC+depression group and DDC+depression+Clomipramine group showed varying degrees of increase.Compared with the DDC+depression group,the ROS content in the muscles of mice in the DDC+depression+Clomipramine group was significantly reduced.Compared with the control group,the serum and muscle KYN levels of mice in the DDC group,DDC+depression group and DDC+depression+Clomipramine group were significantly increased(P<0.05),while KYNA levels were significantly decreased(P<0.05).The concentration of KYN in serum and muscle of mice in the DDC+depression group was significantly higher than that in the DDC group(P<0.05).The concentration of KYN in the serum and muscle of mice in the DDC+depression+Clomipramine group was significantly lower than that in the DDC+depression group(P<0.05).[WTHZ]Conclusion The KYN/KYNA pathway has a regulatory effect on peripheral fatigue in mice with primary biliary cholangitis accompanied by fatigue,and antidepressant therapy can improve muscle fatigue.