Objective To explore the mechanism of Yanggan Anhun Decoction in the treatment of insomnia with liver failing in storing soul type based on network pharmacological methods;To perform animal model validation.Methods The drug components and targets of Yanggan Anhun Decoction were retrieved from TCMSP and BATMAN-TCM databases,and the targets of insomnia with liver failure store soul type were retrieved from GeneCards,NCBI and DisGeNET databases.By constructing a protein-protein interaction(PPI)network and analyzing GO and KEGG pathway enrichment,the signaling pathway of Yanggan Anxin Decoction in treating insomnia with liver failing in storing soul type was determined,and molecular docking was performed between the main active components and core targets.24 SD rats were randomly divided into normal group,model group,TCM group and Western medicine group,with 6 rats in each group.The insomnia model rats with liver failing in storing soul type were constructed by compound multi factor stimulation method,and were given drugs for 14 days.The cognitive memory ability of rats were detected by Morris water maze test;Nissl staining was used to observe the morphology and number of Nissl bodies in the hypothalamus of rats;serum IL-6 and TNF-α contents were detected by ELISA;IL-6 and TNF-α in hypothalamus and Bcl-2,Bax protein expression levels of rats were detected by immunohistochemistry.The relative expressions of p38 MAPK,p-p38 MAPK,Bcl-2,Bax proteins in the hypothalamus of rats were detected by Western blot.Results A total of 301 active components,321 potential targets and 92 key targets were obtained.Key active components such as quercetin,kaempferol,luteolin,wogonin,sebiferic acid and β-sitosterol,as well as core targets such as MAPK,IL6 and TNF were obtained after screening.The key targets mainly focused on various signaling pathways such as TNF signaling pathway,MAPK signaling pathway,PI3K-Akt signaling pathway,dopaminergic synapse,and neuroactive ligand-receptor interaction,etc.Kaempferol,quercetin,digitonin and other flavonoids had good binding activity and stability with MAPK,TNF,IL6 and showed good affinity.The results of animal experiments showed that,compared with the normal group,the cognitive memory ability of the model group decreased(P<0.05),Nissl bodies were stained shallowly,the density was lower,the cell body was shrunk and deformed,the cell nucleus was broken and dissolved,and the serum and hypothalamic IL-6,TNF-α increased(P<0.05),the expressions of p-p38 MAPK/p38 MAPK and Bax in hypothalamus increased(P<0.05),while the expressions of Bcl-2 and Bcl-2/Bax decreased(P<0.05);compared with the model group,the cognitive memory ability of the TCM group and the Western medicine group were improved,the number of Nissl bodies significantly increased,the nucleus was clear,the cell body shrinkage and deformation were improved(P<0.05),the levels of IL-6 and TNF-α in serum and hypothalamus decreased(P<0.05),while the expressions of p-p38 MAPK/p38 MAPK and Bax decreased,and the expression of Bcl-2 and Bcl-2/Bax increased(P<0.05).Conclusion Yanggan Anhun Decoction may play its effects in treating insomnia with liver failing in storing soul type by regulating MAPK,TNF,IL6,Bcl-2,Bax and other core targets,and interfering with p38MAPK signaling pathway.

ObjectiveTo investigate the effects of Guizhi Jia Longgu Mulitang on the expression difference of interleukin-10 （IL-10） and tumor necrosis factor -α （TNF-α） in related organs of insomnia rats with sensory dysfunction dominated by lung and study the mechanism of Guizhi Jia Longgu Mulitang in improving insomnia. MethodSD rats were randomly divided into the blank group， model group， western medicine group， and traditional Chinese medicine （TCM） group， with 10 rats in each group. The rats were deprived of sleep by shallow water environment method in a long platform， and the modeling lasted for 42 d. The blank group and model group were given 0.05 mL·kg^-1 normal saline by gavage， and the western medicine group and TCM group were given drugs during modeling. To be specific， the western medicine group was given 0.105 mg·kg^-1 dexzopiclone tablet by gavage， while the TCM group was given 7 600 mg·kg^-1 Guizhi Jia Longgu Mulitang by gavage， both lasting for 28 days. After successful modeling， the Morris water maze experiment was performed on the 42nd day to detect the motion and spatial memory ability of rats. The levels of IL-10 and TNF-α in serum were detected by enzyme-linked immunosorbent assay （ELISA）. The protein expression of IL-10 and TNF-α in the lung and brain tissue of rats was detected by Western blot. The levels of IL-10 and TNF-α in the lung and brain tissue of rats were detected by immunohistochemistry. ResultCompared with the blank group， the sleep stages non-rapid eye movement ( NREM ) and rapid eye movement ( REM ) of the model group were significantly shortened （P<0.5， P<0.01）， and the wake stage was significantly increased （P<0.01）. The total time and distance of platform exploration were significantly increased （P<0.01）. In the target quadrant （the third quadrant）， the percentage of exploration time and the times of crossing the platform were significantly decreased （P<0.01）. ELISA results showed that the serum IL-10 level was significantly decreased （P<0.01）， and TNF-α level was significantly increased （P<0.01）. The results of Western blot showed that the protein expression of IL-10 in brain and lung tissue of rats was significantly decreased （P<0.01）， and the protein expression of TNF-α was significantly increased （P<0.01）. The results of immunohistochemistry showed that the expression of IL-10 in the brain and lung tissue of rats was significantly decreased （P<0.01）， and that of TNF-α was significantly increased （P<0.01）. Compared with the model group， the NREM stage and REM stage of the western medicine group and the TCM group were significantly increased （P<0.5， P<0.01）， and the wake stage was shortened （P<0.5）. The total time and distance of platform exploration were significantly decreased （P<0.01）. In the target quadrant （the third quadrant）， the percentage of exploration time and the times of crossing the platform were significantly increased （P<0.05， P<0.01）. The serum IL-10 level was significantly increased （P<0.01）， and the serum TNF-α level was significantly decreased according to the ELISA results （P<0.01）. The results of Western blot showed that the protein expression of IL-10 in brain tissue and lung tissue was significantly increased （P<0.01）， and the protein expression of TNF-α was significantly decreased （P<0.01）. The results of immunohistochemistry showed that the expression of IL-10 in brain tissue and lung tissue was significantly increased （P<0.05， P<0.01）， and the expression of TNF-α was significantly decreased （P<0.05， P<0.01）. ConclusionGuizhi Jia Longgu Mulitang can improve the expression of IL-10 and TNF-α in brain and lung tissue of insomnia rats with sensory dysfunction dominated by lung， prolong sleep time， and then improve insomnia. The mechanism may be related to improving the expression level of inflammatory factors.