小檗碱对缺血再灌注小鼠骨骼肌、肾脏中UCP2及线粒体动力学相关蛋白的调节作用  

Study on berberine regulating relationship between UCP2 and mitochondrial dynamics in skeletal muscle and kidney of mice with ischemia-reperfusion

作　　者：张永春 孙林 徐凯 陈绪龙 李昊 张永创 杨清滔 ZHANG Yongchun;SUN Lin;XU Kai;CHEN Xulong;LI Hao;ZHANG Yongchuang;YANG Qingtao(Second Department of Surgery,Hospital of Armed Police Guizhou Corps,Guiyang,Guizhou 550005,China;Department of Administration,Hospital of Armed Police Guizhou Corps,Guiyang,Guizhou 550005,China;Department of Urological Surgery,Affiliated Hospital of Guizhou Medical University,Guiyang,Guizhou 550004,China)

机构地区：[1]武警贵州省总队医院外二科,贵阳550005 [2]武警贵州省总队医院卫勤处,贵阳550005 [3]贵州医科大学附属医院泌尿外科,贵阳550004

出　　处：《重庆医学》2024年第15期2254-2260,共7页Chongqing Medical Journal

基　　金：贵州省卫生健康委员会科学技术基金项目(gzwkj2021-199)。

摘　　要：目的研究小鼠下肢缺血再灌注损伤(IRI)时小檗碱对骨骼肌及肾脏中UCP2表达及线粒体动力学的影响。方法将30只雄性昆明小鼠随机分为阴性对照组、阳性对照组及低、中、高剂量小檗碱干预组(n=6)。各实验组小鼠使用止血带构建双下肢缺血再灌注损伤模型,并腹腔注射不同剂量的小檗碱溶液,阳性对照组使用生理盐水替代。使用苏木素-伊红(HE)染色检测骨骼肌、肾脏病理情况,PCR及Western blot检测线粒体解偶联蛋白2(UCP2)、线粒体分裂蛋白1(FIS1)、线粒体动力蛋白相关蛋白1(DRP1)、线粒体融合蛋白1(Mfn1)、线粒体融合蛋白2(Mfn2)的基因和蛋白表达水平,并检测超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)的变化。结果当小鼠下肢缺血再灌注损伤后,骨骼肌、肾脏均出现炎性细胞浸润,骨骼肌的细胞结构出现损伤性变化;同时,骨骼肌中UCP2、FIS1、DRP1的基因和蛋白表达水平及GSH、SOD水平明显升高(P<0.05),Mfn1、Mfn2的基因和蛋白表达水平及MDA水平明显降低(P<0.05),肾脏中UCP2、DRP1的基因与蛋白表达上升存在差异性(P<0.05)。小檗碱可上调UCP2在骨骼肌的基因表达,以及在肾脏中的蛋白表达(P<0.05),同时,DRP1的基因和蛋白水平在肾组织中受到明显抑制(P<0.05),而在骨骼肌中无明显变化。结论小鼠下肢缺血再灌注损伤导致受损部位出现剧烈的氧化应激损伤,线粒体动力学失衡,并引起肾脏的炎性损害。小檗碱对骨骼肌、肾脏IRI的治疗作用可能是通过抑制氧化应激损伤,其中对肾脏的保护作用还可能与上调UCP2后抑制DRP1表达从而限制线粒体分裂、减缓损伤发展有关。Objective To investigate the effects of berberine on UCP2 expression and the mitochondrial dynamics in skeletal muscle and kidney in mice with ischemia reperfusion injury(IRI)of lower limb.Methods Thirty male Kunming mice were randomly divided into the negative control group,the positive control group,and low,medium and high doses berberine intervention groups.The mice in all experimental groups constructed the ischemia reperfusion injury model of lower limbs by tourniquet,different doses of berberine solution were injected intraperitoneally for intervention,while the ischemia repeating supply group used normal saline for replacement.The HE staining was used to detect the pathological conditions of skeletal muscle and kidney,PCR and Western blot were used to detect the gene and protein expression levels of UCP2,FIS1,DRP1,Mfn1 and Mfn2,and the changes of SOD,MDA and GSH were detected by kit.Results After IRI intervention in the both lower extremities,the inflammatory cell infiltration occurred in both skeletal muscle and kidney,and the cell structure of skeletal muscle showed the damage changes.Meanwhile,the gene and protein expressions levels of UCP2,FIS1 and DRP1 and the levels of GSH and SOD in skeletal muscle were significantly increased(P<0.05),while the gene and protein expression levels of Mfn1 and Mfn2 and the levels of MDA were significantly decreased(P<0.05).The increase of UCP2 and DRP1 gene and protein expression levels in kidney was different(P<0.05).Berberine could up-regulate the UCP2 gene expression in skeletal muscle and the protein expression in skeletal muscle(P<0.05).At the same time,DRP1 gene and protein were significantly inhibited in the kidney tissue(P<0.05),but which in skeletal muscle had no significant change.Conclusion IRI of skeletal muscle of lower extremity in mice leads to severe oxidative stress injury,mitochondrial dynamic imbalance and inflammatory damage of kidney in the injured parts.The therapeutic effect of berberine on skeletal muscle and kidney IRI may be achieved by inhibi

关 键 词：小檗碱 缺血再灌注 骨骼肌 肾脏 UCP2 线粒体动力 

分 类 号：R285.5[医药卫生—中药学]