基于节律调控线粒体信号通路探讨酸枣仁的不同炮制品及不同提取部位对失眠大鼠的作用机制  

Mechanism of different processed products and different extraction parts of Ziziphi Spinosae Semen on insomnia rats based on the rhythm regulation of mitochondrial signaling pathway

作　　者：董珂旭 程世赞 朱月健 王君婷 尹磊 刘国卫 史辑[1,2] 贾天柱 DONG Kexu;CHENG Shizan;ZHU Yuejian;WANG Junting;YIN Lei;LIU Guowei;SHI Ji;JIA Tianzhu(College of Pharmacy,Liaoning University of Traditional Chinese Medicine,Dalian 116600,China;Liaoning TCM Processing Engineering Technology Research Center,Dalian 116600,China;Anhui Jiren Pharmaceutical Co.,Ltd.,Bozhou 236800,China;Anhui Province Traditional Chinese Medicine Extraction Technology Innovation Center,Bozhou 236800,China;Department of Pharmacy,Dalian Public Health Clinical Center,Dalian 116031,China)

机构地区：[1]辽宁中医药大学药学院,辽宁大连116600 [2]辽宁省中药炮制技术产业创新中心,辽宁大连116600 [3]安徽济人药业股份有限公司,安徽亳州236800 [4]安徽省中药提取技术创新中心,安徽亳州236800 [5]大连市公共卫生临床中心药剂科,辽宁大连116031

出　　处：《药学研究》2025年第3期209-216,共8页Journal of Pharmaceutical Research

基　　金：国家自然科学基金项目(No.81874345);沈阳市中青年科技创新人才支持计划项目(No.RC200174);辽宁省自然科学基金资助面上项目(No.2022-MS-223);国家中医药管理局重点实验室研究领域的中医临床疗效提升项目(No.2100222179)。

摘　　要：目的研究酸枣仁不同炮制品及不同提取部位对失眠(DIMS)大鼠的调节作用及作用机制。方法将SD雄性大鼠随机分为空白组,模型组,生酸枣仁总黄酮组、炒酸枣仁总黄酮组、枣叶拌蒸酸枣仁总黄酮组(200 mg·kg^(-1)),生酸枣仁总皂苷组、炒酸枣仁总皂苷组、枣叶拌蒸酸枣仁总皂苷组(200 mg·kg^(-1)),阳性药地西泮组(2.0 mg·kg^(-1)),每组6只。腹腔注射对氯苯丙氨酸(PCPA)建立DIMS大鼠模型。ELISA检测大鼠血清多巴胺(DA)、乙酰胆碱(ACH)、L-谷氨酸(L-Glu)、5羟色胺(5-HT)、γ-氨基丁酸(GABA)水平;RT-PCR检测下丘脑周期蛋白1(Per1)、周期蛋白2(Per2)与脑和肌肉芳烃受体核转录因子样蛋白-1(Bmal1)mRNA表达,Western blotting检测下丘脑线粒体融合蛋白1(Mfn1)、线粒体融合蛋白2(Mfn2)、视神经萎缩相关蛋白1(Opa1)、动力相关蛋白1(Drp1)和线粒体裂变蛋白1(Fis1)蛋白的表达。结果与模型组相比,各给药组均能降低大鼠血清中DA、ACH以及L-Glu含量(P<0.01);均能提升大鼠血清中5-HT含量(P<0.01)及GABA含量,以炒酸枣仁总黄酮组和蒸酸枣仁总皂苷组GABA含量提升作用明显(P<0.01);大鼠下丘脑组织中的Opa1蛋白除炒酸枣仁总皂苷组外各给药组均降低,其中枣叶拌蒸酸枣仁总皂苷组明显降低(P<0.01),Mfn1蛋白除阳性药组和生酸枣仁黄酮组外各给药组均降低,其中炒酸枣仁总黄酮组、枣叶拌蒸酸枣仁总黄酮组、生酸枣仁总皂苷组、枣叶拌蒸酸枣仁总皂苷组明显降低(P<0.01),Mfn2蛋白除炒酸枣仁总皂苷组外各给药组均降低,其中生酸枣仁总黄酮组明显降低(P<0.01),Fis1蛋白各给药组均降低,其中生酸枣仁总黄酮组、炒酸枣仁总黄酮组、枣叶拌蒸酸枣仁总黄酮组、炒酸枣仁总皂苷组明显降低(P<0.01);各给药组大鼠下丘脑组织中的Drp1蛋白相对表达量均升高,以炒酸枣仁总皂苷组升高明显(P<0.01)。结论酸枣仁不同炮制品的总黄酮及总皂苷提取部�Objective To study the regulatory effect and mechanism of different extraction sites of different processed products of Ziziphi Spinosae Semen on disorder of initiating and maintaining sleep(DIMS)rats.Methods SD male rats were randomly divided into blank group,model group,total flavonoids of raw Ziziphi Spinosae Semen group,total flavonoids of fried Ziziphi Spinosae Semen group,total flavonoids of Ziziphus jujuba leaf co-steaming Ziziphi Spinosae Semen group(200 mg·kg^(-1)),total saponins of raw Ziziphi Spinosae Semen group,total saponins of fried Ziziphi Spinosae Semen group,total saponins of Ziziphus jujuba leaf co-steaming Ziziphi Spinosae Semen group(200 mg·kg^(-1)),and positive drug diazepam group(2.0 mg·kg^(-1)),with 6 rats in each group.DIMS rat model was established by intraperitoneal injection of p-chlorophenylalanine.The levels of dopamine(DA),acetylcholine(ACH),L-glutamic acid(L-Glu),5-hydroxytryptamine(5-HT)and γ-aminobutyric acid(GABA)in serum of rats were detected by ELISA.The mRNA expressions of hypothalamic cyclin 1(Per1),cyclin 2(Per2)and brain and muscle aromatic receptor nuclear transcription factor-like protein 1(Bmal1)were detected by RT-PCR.The protein expressions of whole brain mitochondrial fusion protein 1(Mfn1),mitochondrial fusion protein 2(Mfn2),optic atrophy-related protein 1(Opa1),dynamin-related protein 1(Drp1)and mitochondrial fission protein 1(Fis1)were detected by Western blotting.Results Compared with the model group,each administration group could significantly reduce the contents of DA,ACH and L-Glu in serum of rats(P<0.01).The content of 5-HT in serum of rats was significantly increased(P<0.01).Both of them could increase the content of GABA in serum of rats,and the effect of total flavonoids group and total saponins group was obvious(P<0.01).The Opa1 protein in the hypothalamus of rats was decreased in all the administration groups except the total saponins group of fried Ziziphi Spinosae Semen,and the total saponins group of jujube leaves mixed with steamed Ziziphi Spin

关 键 词：酸枣仁 成分 炮制品 失眠 线粒体节律 线粒体信号通路 

分 类 号：R285.5[医药卫生—中药学]