Exploration of the Active Component and Mechanisms of Shengyu Decoction Against Myelosuppression Using Network Pharmacology and in vitro Experimental Validation.

Background: Chemotherapy-induced myelosuppression (CIM) is a common adverse reaction with a high incidence rate that seriously affects human health. Shengyu Decoction (SYD) is often used to treat CIM. However, its pharmacodynamic basis and therapeutic mechanisms remain unclear. Purpose: This study aimed to clarify the active components and mechanisms of SYD in CIM. Methods: LC-QTOF/MS was used to identify the absorbable components of SYD. A series of network pharmacology methods have been applied to explore hub targets and potential mechanisms. Molecular docking was used to identify the binding ability of potential active ingredients and hub targets. Finally, in vitro experiments were performed to validate these findings. Results: In this study, 33 absorbable prototype components were identified using LC-QTOF/MS. A total of 62 possible targets of SYD in myelosuppression were identified. KEGG pathway enrichment analyses showed that some signaling pathways such as PI3K-Akt and HIF-1 may be the mechanisms by which it functions. Among them, we verified the PI3K-Akt pathway. 6 Hub proteins were screened by Protein-protein interaction (PPI) network analysis. Molecular docking results showed that four absorbable components in SYD showed good binding with six Hub targets. The effectiveness of the four predicted compounds and the mechanism were verified in vitro. It has also been shown that the active component could promote the proliferation of bone marrow stromal cells (BMSCs) and block apoptosis of BMSCs, which may be related to the PI3K-Akt pathway. This result is consistent with the network pharmacology approach and molecular docking predictions. Conclusion: Our results provided not only the candidate active component of SYD, but also a new insights into mechanism of SYD in the treatment of CIM.

Combination of ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry and network pharmacology to reveal the mechanism of Shengyu Decoction for treating anemia.

Anemia is a common clinical hematological disease with a high incidence, which seriously affects human health. Shengyu Decoction is often used in the treatment of anemia. However, the pharmacodynamic substance basis and therapeutic mechanism are still unclear, which hinders the comprehensive development and utilization of Shengyu Decoction. In this study, 143 compounds were identified in Shengyu Decoction using high-throughput ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry, 24 of which were absorbed into the blood. Taking these blood-entering ingredients as the research object, we found through network pharmacology research that ferulic acid, calycosin, and astragaloside A can act on AKT1, MAPK1, and MAPK14, and play a role in treating anemia through PI3K-Akt signaling pathway and Pathways in anemia. Finally, it was demonstrated that the active compound could bind to the core target with good affinity by molecular docking. The research shows that Shengyu Decoction has multi-component, multi-target, and multi-channel effects in the treatment of anemia, which provides a basis for the development and clinical application of Shengyu Decoction.

Shengyu Decoction treating vascular cognitive impairment by promoting AKT/HIF-1α/VEGF related cerebrovascular generation and ameliorating MAPK/NF-κB mediated neuroinflammation.

ETHNOPHARMACOLOGICAL RELEVANCE: Shengyu Decoction (SYD), a classical Chinese medicine formula, is good at nourishing blood, promoting blood circulation, and soothe the nerves. SYD can improve cognitive ability. This decoction is suitable for treating vascular cognitive impairment (VCI). however, its active ingredients and possible mechanism have not been investigated. AIM OF THE STUDY: This study was conducted to observe the effects of SYD on improving the cognitive abilities of rats with VCI, to explore its active ingredients and mechanism. MATERIALS AND METHODS: The rats with VCI model were established by bilateral common carotid artery occlusion (BCCAO), and the effects of SYD (5, 2.5 g/kg) on the cognitive abilities of VCI rats were evaluated using the Morris water maze (MWM) and neurological assessment. The pathological changes of hippocampal CA1 were observed by H &E and Nissl staining. The effect of SYD on cerebral blood flow (CBF) was evaluated by Laser Speckle Contrast Imager. The expression of CD31 in the cerebral cortex was measured by immunofluorescence (IF) to evaluate the number of cerebral micro vessels. The levels of IL-6, IL-1ß, and TNF-α in the hippocampus were determined using an ELISA kit, and the active components in the plasma and brain tissues of rats after SYD administration were analyzed using UPLC-Q-TOF-MS/MS. The interaction network of the compound-target pathway was established using the SWISS Target, GO, and DAVID databases. The expression of AKT/HIF-1α/VEGF and p38 MAPK signaling pathway in the brain tissues was determined using western blotting (WB). RESULTS: SYD (2.5, 5 g/kg) significantly improved the cognitive abilities of VCI rats in the MWM and neurological assessment. H&E and Nissl staining showed that SYD significantly ameliorated the pathological hippocampal CA1 area and increased the number of Nissl bodies. The Laser Speckle Contrast Imager showed that the cortical CBF of VCI rats in the SYD group was significantly increased, and the IF results showed that CD31 expression was significantly increased in the SYD group. The ELISA results showed that the contents of IL-6, IL-1ß, and TNF-α in SYD were significantly reduced. A total of 29 compounds were found in the plasma and brain tissues of the rats treated with SYD. Network pharmacology revealed 99 targets for the treatment of VCI. Pathway enrichment analysis showed that the HIF-1 and MAPK signaling pathways might be important for SYD to ameliorate VCI. WB showed that the expressions of AKT, HIF-1α, and VEGF in the brain tissues of rats were significantly increased; in addition, NF-κB and p38 MAPK were significantly reduced in the SYD group. CONCLUSION: SYD can improve the cognitive abilities of VCI rats. The mechanism of action of its active ingredients improves cognitive impairment by affecting the AKT/HIF-1α/VEGF and p38 MAPK/NF-κB signaling pathways, promoting cerebrovascular generation, and ameliorating neuroinflammation.

Content Determination of 4 Indicator Components in Shengyu Decoction Lyophilized Powder by HPLC / 中国药房

OBJECTIVE：To establish the content determin ation method of ferulic acid ，verbascoside，ligustilide and astragaloside in Shengyu decoction lyophilized powder. METHODS ：HPLC method was adopted to determine 4 components in 3 batches of lyophilized powder. The determination of ferulic acid ，verbascoside and ligustilide was performed on Inertsil ODS-SP C 18 column with mobile phase consisted of methanol- 0.1％ phosphoric acid （gradient elution ）at the flow rate of 1.0 mL/min；detector was diode array detector ；detection wavelength was set at 330 nm；column temperature was 30 ℃，the sample size was 10 μL. The determination of astragaloside was performed on Kromasil C 18 column with mobile phase consisted of acetonitrile-water （32∶68，V/ V）；detector was evaporative light scattering detector ；the drift tube temperature wa s 100 ℃，the carrier gas （air）flow rate was 2.5 L/min at the flow rate of 1.0 mL/min；column temperature was 30 ℃，the sample size was 10 μL. RESULTS：The linear ranges of ferulic acid ，verbascoside，ligustilide and astragaloside were 0.050 15-10.03 μg（r＝0.999 8），0.067 80-13.56 μg（r＝ 0.999 9），0.057 30-11.46 μg（r＝0.999 5），1.128-11.28 μg（r＝0.999 3），respectively. The detection limits were 2.12×10－4，1.30× 10－3，8.02×10－4，1.09×10－3 μg，respectively. The limit of quantification were 7.43×10－4，3.87×10－3，2.34×10－3，3.36×10－3 μg， respectively. RSDs of precision ，stability（12 h）and reproducibility tests were all lower than 2％（n＝6）. Average recovery rates were 99.6％（RSD＝0.83％，n＝6），100.9％（RSD＝1.07％，n＝6），98.8％（RSD＝0.84％，n＝6）and 101.3％（RSD＝0.99％， n＝6），respectively. The contents of ferulic acid ，verbascoside，ligustilide and astragaloside in 3 batches of samples were 1.225-1.248， 0.413-0.424， 0.325-0.332， 0.394-0.404 mg/g， respectively （RSDs among batches were lower than 1.5％ ）. CONCLUSIONS：Established method is stable ，reproducible，rapid and accurate for the content determination of ferulic acid ， verbascoside， ligustilide and astragaloside in Shengyu

Common and unique mechanisms of Chinese herbal remedies on ischemic stroke mice revealed by transcriptome analyses.

ETHNOPHARMACOLOGICAL RELEVANCE: Four traditional Chinese herbal remedies (CHR) including Buyang Huanwu decoction (BHD), Xuefu Zhuyu decoction (XZD), Tianma Gouteng decoction (TGD) and Shengyu decoction (SYD) are popular used in treating brain-related dysfunction clinically with different syndrome/pattern based on traditional Chinese medicine (TCM) principles, yet their neuroprotective mechanisms are still unclear. MATERIALS AND METHODS: Mice were subjected to an acute ischemic stroke to examine the efficacy and molecular mechanisms of action underlying these CHR. RESULTS: CHR treatment significantly enhanced the survival rate of stroke mice, with BHD being the most effective CHR. All CHR were superior to recombinant tissue-type plasminogen activator (rt-PA) treatment in successfully ameliorating brain function, infarction, and neurological deficits in stroke mice that also paralleled to improvements in blood-brain barrier damage, inflammation, apoptosis, and neurogenesis. Transcriptome analyses reveals that a total of 774 ischemia-induced probe sets were significantly modulated by four CHR, including 52 commonly upregulated genes and 54 commonly downregulated ones. Among them, activation of neurogenesis-associated signaling pathways and down-regulating inflammation and apoptosis pathways are key common mechanisms in ischemic stroke protection by all CHR. Besides, levels of plasma CX3CL1 and S100a9 in patients could be used as biomarkers for therapeutic evaluation before functional recovery could be observed. CONCLUSION: Our results suggest that using CHR, a combinatory cocktail therapy, is a better way than rt-PA for treating cerebral ischemic-associated diseases through modulating a common as well as a specific group of genes/pathways that may partially explain the syndrome differentiation and treatment principle in TCM.

Improvement in the neural stem cell proliferation in rats treated with modified "Shengyu" decoction may contribute to the neurorestoration.

ETHNOPHARMACOLOGICAL RELEVANCE: "Shengyu" decoction, a traditional Chinese medicine, has been used to treat diseases with deficit in "qi" and "blood". The modified "Shengyu" decoction (MSD) used in the present study was designed to treat traumatic brain injury (TBI) on the basis of the "Shengyu" decoction, in which additional four herbs were added. Many ingredients in these herbs have been demonstrated to be effective for the treatment of brain injury. The present study was performed to evaluate the neurorestorative effect and the underlying mechanisms of MSD on the rat brain after a TBI. MATERIALS AND METHODS: TBI was induced in the right cerebral cortex of adult rats using Feeney's weight-drop method. Intragastrical administration of MSD (1.0 ml/200 g) was begun 6h after TBI. The neurological functions and neuronal loss in the cortex and hippocampus were determined. The levels of nerve growth-related factors GDNF, NGF, NCAM, TN-C, and Nogo-A and the number of GFAP(+)/GDNF(+), BrdU(+)/nestin(+), BrdU(+)/NeuN(+) immunoreactive cells in the brain ipsilateral to TBI were also measured. Moreover, the influences of MSD on these variables were observed at the same time. RESULTS: We found that treatment with MSD in TBI rats ameliorated the neurological functions and alleviated neuronal loss. MSD treatment elevated the expression of GDNF, NGF, NCAM, and TN-C, and inhibited the expression of Nogo-A. Moreover, MSD treatment increased the number of GFAP(+)/GDNF(+), BrdU(+)/nestin(+), and BrdU(+)/NeuN(+) immunoreactive cells in the cortex and hippocampus. CONCLUSION: The present results suggest that MSD treatment in TBI rats could improve the proliferation of neural stem/progenitor cells and differentiation into neurons, which may facilitate neural regeneration and tissue repair and thus contribute to the recovery of neurological functions. These effects of modified "Shengyu" decoction may provide a foundation for the use of MSD as a prescription of medicinal herbs in the traditional medicine to treat brain injuries in order to improve the neurorestoration.

The neuroprotective effect of modified "Shengyu" decoction is mediated through an anti-inflammatory mechanism in the rat after traumatic brain injury.

ETHNOPHARMACOLOGICAL RELEVANCE: "Shengyu" decoction, a traditional Chinese medicine, has been used to treat diseases with deficit in "qi" and "blood" induced frequently by profound loss of blood or by long sores with heavy pus, in which a potential anti-inflammatory effect is implied. The modified "Shengyu" decoction (MSD) used in the present study was designed on the basis of the "Shengyu" decoction, additional four herbs were added in. Many ingredients in these herbs have been demonstrated to be anti-inflammatory and thus MSD may be used for the treatment of traumatic brain injury (TBI). To evaluate the neuroprotective effect and the underlying mechanisms of MSD on the rat brain after TBI. MATERIALS AND METHODS: TBI was induced in the right cerebral cortex of male adult rats using Feeney's weight-drop method. The rats were administered a gavage of MSD (0.5, 1.0 or 2.0 ml/200 g) 6h after TBI. The neurological functions, brain water content, contusion volume, and neuron loss were determined. The levels of TNF-α, IL-1ß, IL-6, and IL-10 and the number of GFAP- and Iba1-positive cells in the brain ipsilateral to TBI were also measured. Moreover, the influence of MSD on these variables was observed at the same time. RESULTS: The neurological deficits, brain water content, and neuron loss were significantly reduced after 1.0 or 2.0 ml/200 g of MSD treatment but not after 0.5 ml/200 g. In addition, treatment with MSD (1.0 ml/200 g) significantly increased the level of IL-10 and reduced the level of TNF-α and IL-1ß and the number of GFAP- and Iba1-positive cells after TBI. However, the contusion volume of brain tissue and the expression of IL-6 were not significantly changed. CONCLUSION: MSD may be a potential therapeutic for the treatment of TBI because MSD alleviated secondary brain injury induced by TBI. In addition, MSD inhibited the inflammatory response through reducing the expression of inflammatory cytokines and the activation of microglial cells and astrocytes in the brain tissue of rats after TBI. Therefore, a potential anti-inflammatory mechanism of the "Shengyu" decoction was confirmed, which may be one of the main reasons of "Shengyu" decoction used to treat diseases with obvious inflammatory responses.

Experimental Study on Effect of Shengyu Decoction and Its Decomposition on Rats Red Cell EPO / 浙江中医药大学学报

[Objective]To explore the functional mechanism and compatibility of Shengyu Decoction notifying blood.[Method] Take60Coy ray radiation to induce blood deficiency mice model of radioactive injury,observe and compare the effect of the decoction and its decomposition on recovery of hemopoiesis injury and regulating EPO level,embodying the main function of drugs of reinforcing essence and notifying blood EPO level of mice in blood deficiency is the effective link of improving anemia and promoting recovery of hemopoiesis;its combination of notifying Qi,essence and producing blood embodies the whole efficacy of mutual coordination and control.