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

DONG Kexu; CHENG Shizan; ZHU Yuejian; WANG Junting; YIN Lei; LIU Guowei; SHI Ji; JIA Tianzhu

doi:10.13506/j.cnki.jpr.2025.03.001

Abstract：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-chlorophe-nylalanine.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 fu-sion protein 2(Mfn2),optic atrophy-related protein 1(Opa1),dynamin-related protein 1(Drp1)and mitochondrial fis-sion 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 Spinosae Semen was significantly decreased(P＜0.01).The Mfn1 protein was decreased in all the administration groups except the positive drug group and the flavonoids group of raw Ziziphi Spinosae Semen,and the total flavonoids group of fried Ziziphi Spinosae Semen,the total saponins group of raw Ziziphi Spinosae Semen and the total saponins group of jujube leaves mixed with steamed Ziziphi Spinosae Semen were significantly decreased(P＜0.01).The Mfn2 protein was decreased in all the administration groups except the total saponins group of fried Ziziphi Spinosae Semen,and the total flavonoids group of raw Ziziphi Spinosae Semen was significantly decreased(P＜0.01).Fis1 protein was decreased in each administration group,among which the total flavonoids group of raw jujube kernel,the total flavonoids group of fried jujube kernel,the total flavonoids group of jujube leaf mixed with steamed jujube kernel and the total saponins group of fried jujube kernel were significantly decreased(P＜0.01);The relative expression of Drp1 protein in hypothalamus tissue of rats in each administration group increased,and the total saponins group increased signifi-cantly(P＜0.01).Conclusion The total flavonoids and total saponins extracted from different processed products of Ziziphi Spinosae Semen play an anti-insomnia role by regulating the level of neurotransmitters and regulating the mitochondrial rhythm through the Mfn1,Mfn2,Opa1,Drp1 and Fis1 signaling pathways.