Levodopa-induced dyskinesia (LID) is a side effect of Parkinson's disease (PD) treated with L-3,4-dihydroxyphenylalanine (L-DOPA). Nardosinone is the major active component of the dried root and rhizome of Nardostachys jatamansi DC, which is effective for treating PD. The purpose of this study was to explore the effects of nardosinone on LID and the underlying mechanism involving the microbiota-gut-brain axis. A PD rat model was established with rotenone. Behavioral and immunohistochemical staining experiments were used to investigate the synergistic effect of nardosinone on L-DOPA in PD rats. Then, the LID model was established by using high-dose L-DOPA and benserazide, and the effect of nardosinone on LID was evaluated by abnormal involuntary movement scale (AIMS) and western blotting (WB). Hematoxylin‒eosin staining (on intestinal tissues), 16 S rRNA (on gut microbiota), and ELISA (on serum and brain tissues) were used to investigate the mechanism of action of nardosinone in the treatment of LID. Behavioral tests revealed that nardosinone increased the ability of L-DOPA to treat forelimb dysfunction and autonomic activity disorders in PD, and immunohistochemical staining experiments revealed that nardosinone exerted a protective effect on dopaminergic neurons as PD-induced neuronal damage was reversed, which complements the therapeutic effect of L-DOPA. AIMS and WB analyses indicated that high-dose L-DOPA induced LID, but nardosinone exerted an effect in reducing LID by not only improving rats' abnormal involuntary movements but also downregulating ∆FosB expression. Analysis of 16 S rRNA revealed that nardosinone can regulate the intestinal flora in rats with LID. In addition, nardosinone can protect the colonic structure and reduce intestinal permeability. Moreover, nardosinone can reduce the expression of inflammatory factors in the colon and striatum of rats with LID and improve intestinal inflammation and neuroinflammation. Nardosinone can improve the effect of L-DOPA on ameliorating motor function in PD rats while alleviating LID, and its mechanism for improving LID may be related to regulating intestinal flora balance, repairing intestinal barrier integrity, and inhibiting inflammatory responses via the microbiota-gut-brain axis.

Background Hepatocellular carcinoma (HCC) accounts for the majority of primary liver cancers and is associated with high incidence rate and high mortality rate. Cremastrae pseudobulbus pleiones pseudobulbus (CPPP) is commonly used in the clinical treatment of HCC, and our preliminary studies have identified four phenanthrenes and one bibenzyl as its primary active components. Purpose This study aimed to establish an extraction and purification process for the active components of CPPP, obtain its purified extract (EPC), and further elucidate the anti-tumor effects of EPC against HCC and its underlying mechanisms. Methods Orthogonal design experiment and macroporous resin chromatography were employed to establish the optimal extraction and purification conditions for EPC. The anti-HCC effects of EPC were evaluated through a comprehensive series of assays including CCK-8 assay, EdU incorporation, and wound healing in vitro, alongside a mouse xenograft model in vivo. The mode of cell death was determined using specific inhibitors, flow cytometry, and Western blot analysis. Mitochondrial morphology and function were assessed using transmission electron microscopy, ROS, ATP and Ca2+ assay kits. The molecular mechanisms were elucidated through thermal proteome profiling, and western blot analysis, with subsequent verification using specific target and pathway activators. Results The content of 5 active components in the purified EPC was 4.74 times higher than that before purification, and its inhibitory effect on HCC cells viability was significantly enhanced. Pharmacological experiments confirmed that EPC could inhibit the proliferation and migration of HCC cells as well as the growth of HCC tumors. Furthermore, EPC induced apoptosis by impairing mitochondrial morphology and function through increasing the protein expression levels of Bax/Bcl-2, cleaved Caspase-3, and cleaved Caspase-9 proteins. Mechanistic analysis revealed that this anti-HCC effect was largely dependent on suppression of the KRAS/PI3K/AKT/mTOR pathway by EPC. Activation of either KRAS protein or the PI3K/AKT/mTOR pathway attenuated the therapeutic efficacy of EPC. Conclusions EPC effectively alleviates HCC progression by predominantly triggering intrinsic, mitochondria-related apoptosis, which is mediated through the suppression of the KRAS/PI3K/AKT/mTOR signaling pathway. Our findings provide evidence for the pharmacological basis of CPPP and identify EPC as a promising purified lead extract warranting further pharmacokinetic and safety evaluation for HCC.

Levodopa (L-DOPA) is the mainstay treatment for Parkinson’s disease (PD), but its long-term use can cause adverse effects. Nardosinone enhances L-DOPA efficacy, yet the underlying mechanism is unclear. This study investigated whether the gut microbiota is associated with this synergism in a rotenone-induced PD rat model. Coadministration of nardosinone and L-DOPA increased L-DOPA and dopamine levels in the feces, plasma, and striatum of normal rats but not in antibiotic-treated rats with perturbed gut microbiota, suggesting a microbiota-correlated effect. Fecal microbiota transplantation from donors receiving the combination therapy alleviated motor deficits and dopaminergic neuron loss in recipient PD rats. This combination was associated with upregulated expression of tyrosine hydroxylase, DOPA decarboxylase, and their cofactors tetrahydrobiopterin and vitamin B6, as well as increased PINK1 and Parkin levels. It also reshaped the gut microbiota composition, enhanced intestinal and blood–brain barrier tight junction protein expression, and reduced c-Jun N-terminal kinase protein expression and neuroinflammation. In conclusion, these findings indicate that nardosinone coadministration is associated with alterations in the gut microbiota, improved barrier integrity markers, attenuated inflammation, and enhanced therapeutic effects of L-DOPA in this PD rat model. Definitive mechanistic pathways underlying the observed associations remain to be established.

Based on an early-stage Parkinson's disease(PD) rat model, this study employed 16S rRNA sequencing and untargeted metabolomics to investigate the action mechanism of Tianma Gouteng Yin(TGY) in treating early-stage PD. The early-stage PD rat models were established by subcutaneous injection of rotenone for seven days and were randomly divided into a control group, a model group, a positive drug group(levodopa, 50 mg·kg~(-1)), and TGY groups with low, medium, and high doses(9.82, 19.64, and 39.28 g·kg~(-1)), with intragastric administration for seven days during modeling. Behavioral indicators of rats(open field, inclined plane, and pole tests) were determined. The pathological morphology of brain tissue and the expressions of tyrosine hydroxylase(TH) and α-synuclein(α-syn) were detected. The levels of neurotransmitters including dopamine(DA), 5-hydroxytryptamine(5-HT), 3,4-dihydroxyphenylacetic acid(DOPAC), levodopa, and homovanillic acid(HVA) were measured. The inflammatory factors including tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), and interleukin-1β(IL-1β) were detected. 16S rRNA sequencing and untargeted metabolomics were conducted on colonic contents to explore the pharmacodynamic effects of TGY and its regulatory mechanisms on gut microbiota and metabolism. RESULTS:: show that TGY can significantly improve motor dysfunction in early-stage PD rats, increase the number of TH-positive cells, inhibit the abnormal aggregation of α-synuclein(α-syn), up-regulate the levels of neurotransmitters, reduce the levels of inflammatory factors in the colon and striatum, thereby exerting a neuroprotective effect. Gut microbiota analysis reveals that TGY can reverse the trends of reduced α-diversity of gut microbiota, increased Firmicutes/Bacteroidetes(F/B) ratio, and increased abundance of pro-inflammatory genera(e.g., Ruminococcus), restore the abundance of beneficial genera(e.g., Bacteroides), and reshape the gut microbiota structure. Metabolomic analysis demonstrates that TGY intervention significantly reverses the differential metabolites in the model group, mainly involving energy and amino acid metabolic pathways such as pyruvate metabolism, β-alanine metabolism, and aminoacyl-tRNA biosynthesis. In conclusion, TGY may exert therapeutic effects on early-stage PD by regulating the axis of microbiota, inflammation, and neuronal injury, with energy and amino acid metabolism as the hub, through these metabolic pathways.

Parkinson's disease (PD) is a progressive disease that requires effective staging management. The role of intestinal microbiota in PD has been studied, but its changes at different stages are not clear. In this study, metaanalysis, bioinformatics analysis and in vivo simulation were used to explore the intestinal microbiota distribution of PD patients and models at different stages. Two PD models at different stages were established in rotenone-treated rats and MPTP-induced mice. The differences in the intestinal microbiota among the different stages of PD patients or models were compared and analyzed. There were significant differences between PD patients and controls, including analysis, we revealed significant differences between PD patients at different stages and controls, including etc. Through meta-analysis, we found that Actinobacteriota and Erysipelotrichaceae had significantly increased in the chronic MPTP model, while Prevotellaceae had significantly decreased. PD rats and mice presented significant damage to motor function, coordination, autonomous activity ability and gastrointestinal function, and the damage in the late group was greater than that in the early group. There were significant differences in intestinal microbiota between PD patients or models at different stages and the control groups. In the early stage, the dominant microbiota are Akkermansia, Alistipes, Anaerotruncus, Bilophila, Rikenellaceae, Verrucomicrobia and Verrucomicrobiae, whereas in the late stage, the dominant microbiota are Actinobacteriota and Erysipelotrichaceae. These differences can lay a foundation for subsequent research on the treatment and mechanism of PD at different stages.

ABSTRACTPanax notoginseng (P. notoginseng) is one of the most famous natural medicines and widely used to promote blood circulation in health care. However, the active component group of P. notoginseng for activating blood is not clear. We aim to screen and validate the pharmacodynamic component group (PCG), which could exert the same blood‐activating effect as P. notoginseng. To clarify the active components, the chemical components were determined by liquid chromatography‐tandem mass spectrometry, and the fingerprint of P. notoginseng was established. Twenty candidate active monomers were selected through the spectrum–effect relationship analysis. Eleven active monomers, including Ginsenoside Rg1, Rb1, Rd, F1, Rh1, Rg2, Rb2, Rg3, and Rk1 and Notoginsenoside R1 and R2, were screened out as the PCG through validation by platelet aggregation test. Among them, the antiplatelet aggregation activity of Ginsenoside Rh1 was directly confirmed for the first time. The active component group could exert similar efficacy to the P. notoginseng extract in vitro and in vivo through the validation of in vitro platelet aggregation test and the rats with cerebral ischemia. This study laid the foundation for the quality evaluation of P. notoginseng and provided a reference for the research on the material basis of the pharmacodynamics of other Chinese herbs.

Natural products (NPs) make a major contribution to drug development, offering a huge molecule pool for drug leads. Nevertheless, the pharmaceutical industry and academy have declined their enthusiasm to NPs research since the great challenges in elucidating the complex component and intricate mechanism of NPs. Here, we introduce an efficient fragment-based target research (FBTR) approach for pharmacology study and optimization of NPs. Focusing on the core fragment within the molecules of NPs, we screen the outstanding activity that be triggered, and corresponding target. Finally, drug optimization was carried out around the molecules that obtaining the activity-related core fragment and verified both in vitro and in vivo. With this approach, we obtained an optimized NPs named Erigeron breviscapus polyphenols (EBP) with definite target. After optimization, EBP plus (EBPP) not only trigger immunogenic cell death (ICD) of glioblastoma (GBM) cells effectively by targeting to Cys105 amino acid site of Fas-associating protein with a novel death domain (FADD) protein, but also prolong the survival of GBM mice by an average of 17.6 days. Significantly, our investigation presents an approach for addressing challenges in NPs development and opening up new opportunities for drug discovery. Our findings demonstrate the utility of FBTR in exploring the function of NPs, revealing the target, and advancing drug optimization for stronger clinical translation.

Background:Pseudobulbus Cremastrae seu Pleiones (Shancigu), a traditional Chinese medicine (TCM), has been extensively used in clinical practice for the treatment of various tumors, particularly liver cancer. Shancigu is classified into two commercial specifications-"Maocigu" and "Bingqiuzi"-which exhibit significant differences in appearance, chemical composition, and price, posing challenges for the quality control of medicinal materials. Purpose:The aim of this study was to clarify the quality evaluation indicators based on the anti-liver cancer active components in Shancigu and to establish a reliable quality evaluation method to preliminarily assess the quality of Shancigu from different commercial specifications and production areas. Methods:Twenty-six batches of Shancigu samples were collected. High-performance liquid chromatography (HPLC) was used to establish fingerprint spectra. In vitro anti-liver cancer pharmacological effect indicators were analyzed using the CCK-8 assay and scratch wound healing assays. Through spectrum-effect relationship analysis, serum pharmacochemistry analysis, and in vitro/in vivo anti-liver cancer activity evaluation, the effective component combinations of Bingqiuzi and Maocigu were identified and validated. Gray relational analysis (GRA) and the technique for order preference by similarity to ideal solution (TOPSIS) were subsequently applied to assess the quality of Shancigu based on their different specifications and origins. Results:Eleven key anti-liver cancer active components from Shancigu were screened and confirmed, namely, malic acid, citric acid, 2-isobutylmalic acid, gastrodin, batatasin III, 2-p-hydroxybenzyl-5,3'-dihydroxy-3-methoxybibenzyl, coelonin, 1-p-hydroxybenzyl-2,7-dihydroxy-4-methoxyphenanthrene, blestriarene A, blestriarene B, and monbarbatain A. These components are present in Bingqiuzi and Maocigu in different proportions, and the anti-liver cancer pharmacological effects of the effective component combinations were found to be equivalent to those of the original materials, both in vitro and in vivo. These 11 components can be used as indicators for evaluating the quality of Shancigu. Quality evaluations revealed no significant differences between Bingqiuzi and Maocigu. For Bingqiuzi, medicinal materials produced in Guizhou and Yunnan were of better quality; for Maocigu, those from Guizhou and Sichuan were superior. Conclusion:In this study, we established quality evaluation criteria for Shancigu and developed an innovative method to comprehensively assess the quality of Shancigu from different commercial specifications and production regions. By integrating component analysis with anti-liver cancer activity assessment, this research provides a valuable reference for the quality evaluation of other Chinese medicinal materials.

Background: The roots and rhizomes of Nardostachys jatamansi DC. are reported to be useful for the treatment of Parkinson's disease (PD). Previous research has also shown that Nardosinone, the main active component isolated from Nardostachys jatamansi DC., exhibits the potential to treat PD. Aim of the study: To investigate how the effects of Nardosinone could assist levodopa in the treatment of PD, how this process changes the intestinal flora, and to explore the effective forms of Nardosinone in the intestinal flora. Material and methods: We used behavioral experiments, and hematoxylin-eosin staining and immunohistochemical staining, to investigate the effects of a combination of Nardosinone and levodopa on rotenone-induced PD rats. In addition, we used LC/MS-MS to determine the levels of levodopa, 5-hydroxytryptamine, dopamine and its metabolite 3, 4-dihydroxyphenylacetic acid, and homovanillic acid, to investigate the effect of the intestinal flora on co-administration in the treatment of PD. LC/MS-MS was also used to detect the metabolites of Nardosinone on the gastrointestinal tract and intestinal flora. Results: The behavioral disorders and neuronal damage associated with PD were significantly improved following the co-administration. Analysis also revealed that the co-administration increased the levels of five neurotransmitters in the striatum, plasma and feces. In vitro experiments further demonstrated that the levels of dopamine and levodopa were increased in the intestinal flora. In total, five metabolites of Nardosinone were identified. Conclusion: Our findings indicate that Nardosinone and its metabolites might act as a potential adjutant to enhance the efficacy of levodopa via the intestinal flora, thus expanding the therapeutic potential of the combination of Chinese and Western medicine as a treatment method for PD.

Cremastrae Pseudobulbus Pleiones Pseudobulbus (CPPP) is derived from the dried pseudobulb of the orchid family plants Cremastra appendiculata (D.Don) Makino, Pleione bulbocodioides (Franch.) Rolfe, or Pleione yunnanensis Rolfe, and has the properties of clearing heat, detoxification, resolving phlegm, and dispersing nodules. It is frequently used for the treatment of various malignant tumors in clinical practice, especially lung cancer. CPPP is divided into two commercial specifications in the market, Maocigu (MCG) and Bingqiuzi (BQZ). However, owing to a lack of appropriate research strategies, the active ingredients and molecular mechanisms involved have not yet been clarified. This study intended to discover the combination of effective anti-lung-cancer ingredients in CPPP and explore their potential mechanisms of action. In this study, UHPLC-MS fingerprints of MCG and BQZ were established separately. Inhibitory effects on the proliferative viability and migratory ability of A459 and H1299 cells were evaluated as pharmacodynamic indicators. GRA and BCA were used to determine spectrum–effect relationships. Next, the identification and analysis of components of drug-containing serum were performed using UHPLC-Q-Exactive Orbitrap MS. Then, the results of the two analyses were combined to jointly screen out the anti-lung-cancer candidate active monomers of CPPP, and their in vitro activities were verified. Afterward, all effective ingredient combinations of MCG (MCGC) and BQZ (BQZC) were prepared according to their contents in the original medicinal materials. Their anti-lung-cancer activities in vitro and in vivo were compared and verified. Finally, we used the human lung cancer cell line A549 and the Lewis tumor xenograft model to investigate how BQZC would influence autophagy and apoptosis processes and the mechanisms involved. Overall, 11 predominant anti-lung-cancer active ingredients from CPPP were screened. Next, MCGC and BQZC were prepared according to their contents in the original medicinal materials, respectively, and their anti-tumor effects were equivalent to those of the original materials in vitro and in vivo. We found that BQZC could inhibit lung cancer cell growth and induce protective autophagy and apoptosis in lung cancer cells by activating the AMPK–mTOR–ULK1/BMF signaling pathway. These results provide important evidence for the clinical application and deep development of CPPP against tumors.

Objective To investigate the effect of 80% ethanol extract of Nardostachys jatamansi（NJ）on dyskinesia in Parkinson’s（PD）rats, and explore the mechanism of NJ against Parkinson’s disease through the “gut-brain axis”. Methods The rotenone was used to induce PD rats model, rats were set up as blank control group, model group, levodopa（50 mg/kg）group and NJ low-, mediumand high-dose（0.41, 0.62, 0.93 g/kg）groups. To screen optimal dose of 80% ethanol extract of NJ for anti-PD movement disorders,the movement disorders of PD rats were investigated by open field test and inclined plate test, the striatal DA content was measured by ELISA. Immunohistochemical staining was used to investigate the expression of tyrosine hydroxylase（TH）, a characteristic marker of dopamine（DA）neurons in the substantia nigra of rat midbrain; ELISA was used to investigate the contents of DA, 5-hydroxytryptamine（5-HT）and its metabolites 3,4-dihydroxyphenylacetic acid（DOPAC）, homovanillic acid（HVA）and 5-hydroxyphenylacetic acid（5-HIAA）in colon and striatum of rats; Expressions of enteric glial cell biomarker glial fibrillary acidic protein（GFAP）and microglia surface marker ionized calcium binding adapter molecule 1（Iba-1）from colon and striatum were detected by Western blotting. Results Compared with model group, NJ could improve the motor dysfunction of PD rats to varying degrees, and NJ medium-dose group had the best effect, which significantly improved the total distance of open field test and angle of stay of inclined plate test（P ＜ 0.001）, significantly increased DA content in striatum and TH positive expression in substantia nigra of PD rats（P ＜ 0.01, 0.001）. After administration of NJ, the decreasing trend of DA, 5-HT and its metabolites DOPAC, HVA and 5-HIAA in colon and striatum of PD rats were reversed to varying degrees（P ＜ 0.05, 0.01, 0.001）, and DA metabolic rate was inhibited（P ＜0.05）; NJ significantly reduced the expressions of GFAP and Iba-1 in colon and striatum（P ＜ 0.001）, inhibited the activation of enteric glial cells, glial cells and microglia. Conclusion NJ can significantly improve the motor dysfunction of PD rats, and its mechanism may be related to that inhibiting the excessive activation of enteric glial cells and glial cells to prevent the development of neuroinflammation, and inhibiting the metabolic process of neurotransmitters such as DA and 5-HT in colon and striatum to increase their content. This study verifies the theory that NJ exerts its anti-PD effect through the “gut-brain axis”, and can provide new ideas for the research on anti-PD of traditional Chinese medicine that attributed to the spleen and stomach meridians.

BackgroundAlzheimer’s disease (AD) is a multifactorial neurodegenerative condition. The search for multi-target traditional Chinese medicines or ingredients for treating AD has attracted much attention. Corydalis rhizome (CR) is a traditional Chinese medicine. Its main components are alkaloids, which have therapeutic effects that can potentially be used for treating AD. However, no systematic study has been conducted to explore the anti-AD efficacy of CR, as well as its active compounds and mechanisms of action.ObjectiveThe present study aimed to clarify CR’s active constituents and its pharmacological mechanisms in treating AD.MethodsA D-galactose & scopolamine hydrobromide-induced AD mouse model was used and CR was administered orally. The prototypical alkaloid components were identified in the serum. The core components, key targets, and possible mechanisms of action of these alkaloids were revealed through network pharmacology. Molecular docking of the key target was performed. Finally, the mechanism was validated by lipopolysaccharide (LPS)-induced activation of BV2 microglia.ResultsThe results showed that CR improved anxiety-like behavior, spatial and non-spatial recognition, and memory capacity in AD mice. It also achieved synergistic AD treatment by modulating neurotransmitter levels, anti-neuroinflammation, and anti-oxidative stress. The core components that enhance CR’s efficacy in treating AD are protoberberine-type alkaloids. The CR may induce the polarization of LPS-activated BV2 microglia from phenotype M1 to M2. This is partially achieved by modulating the IL-6/JAK2/STAT3 signaling pathway, which could be the mechanism by which CR treats AD through anti-inflammation.ConclusionThe present study provided a theoretical and experimental basis for the clinical application of CR in treating AD. It also provides information that aids the secondary development, and precise clinical use of CR.

目的 建立山慈菇的HPLC指纹图谱,对其特征成分进行含量测定并结合化学计量学分析方法分析不同商品规格山慈菇(冰球子、毛慈菇)化学成分的差异,为山慈菇药材的质量控制提供参考.方法 优化山慈菇药材的提取方法,应用HPLC法分别建立冰球子和毛慈菇指纹图谱,并对特征成分进行含量测定.采用相似度评价、聚类分析(cluster analysis,CA)、主成分分析(principalcomponent analysis,PCA)、正交偏最小二乘法判别分析(orthogonal partial least squares discriminant analysis,OPLS-DA)、Fisher 线性判别分析(fisher linear discrimination analysis,FLDA)对特征成分进行数据分析.结果 建立了冰球子和毛慈菇HPLC指纹图谱,分别匹配了 46个和43个共有峰,共标示出17个色谱峰,指认出天麻素、loroglossin、dactylorhin A、白芨苷、山药素Ⅲ、白芨联菲A、白芨联菲B、卷瓣兰蒽8个色谱峰并同时建立了上述除天麻素以外7个成分的HPLC含量测定方法.CA与PCA结果可将2种不同商品规格山慈菇样品明显区分.OPLS-DA分析结合含量测定Fisher判别分析结果表明,2种不同商品规格的山慈菇样品间存在显著性差异的5种成分是loroglossin、dactylorhin A、白芨苷、白芨联菲B、卷瓣兰蒽(P＜0.001).结论 所建立的HPLC指纹图谱结合CA、PCA、OPLS-DA分析及多成分含量测定能客观、全面、有效地确定不同商品规格山慈菇中主要成分的差异,可为山慈菇质量评价体系的完善提供依据.

Ethnopharmacological relevance: Panax Notoginseng (PN) can disperse blood stasis, hemostasis, and detu-mescence analgesic, which can be used for hemoptysis, hematemesis and another traumatic bleeding, and it is known as "A miracle hemostatic medicine". Studies show that the chemical composition of PN is relatively comprehensive, however, its hemostatic active ingredients have not been fully clarified.Aim of study: This study aimed to clarify the hemostatic effective components group (HECG) of PN, provide a foundation for the assessment of PN's quality and its comprehensive development, and for further studies on the pharmacodynamic material basis of other Traditional Chinese Medicines (TCMs).Materials and methods: UPLC-MS was used to establish the fingerprint and identify the common peaks in 44 batches of PN extracts (PNE). In addition, the plasma recalcification time and in vitro coagulation time were measured. For spectrum-effect analysis, bivariate correlation analysis (BCA) and partial least squares regression analysis (PLSR) were used to screen the hemostasis candidate active monomers of PN. The monomers were prepared by combining several preparative chromatography techniques. The efficacy was verified by plasma recalcification time, in vitro coagulation time, and a rat model of gastric hemorrhage.Results: A total of 30 common peaks and hemostatic efficacy indexes of 44 batches of PNE were obtained. A total of 18 components were positively correlated with the comprehensive coagulation index by two statistical methods. Six and eleven monomers were obtained respectively by chromatographic preparation and procurement, and one monomer was eliminated due to preparation difficulty and other reasons. Seven active monomers with direct hemostatic effect and one active monomer with synergistic hemostatic effect were screened through plasma recalcification time, and their combinations were used as candidate HECG for hemostatic effect verifi-cation. The results of in vitro experiments showed that plasma recalcification time and in vitro coagulation time were significantly reduced (P < 0.05) in the HECG group, compared to the PNE group. The results of in vivo experiment also indicated that the hemostatic effect of HECG was comparable to that of PNE and PN powder. Conclusion: The composition and efficacy of the HECG of PN were screened and verified using the spectral correlation method and in vivo and in vitro efficacy verification; the HECG included Dencichine, Ginsenoside Rg1, Ginsenoside Rd, Ginsenoside Rh1, Ginsenoside F1, Notoginsenoside R1, Notoginsenoside Ft1 and Noto-ginsenoside Fe. These results laid a foundation for the quality evaluation of PN and provided a reference for the basic research of pharmacodynamic material basis of other TCMs.

Cerebral ischemic stroke is a common neuron loss disease that is caused by the interruption of the blood supply to the brain. In order to enhance the CIS outcome, both identifying the treatment target of ischemic brain damage in the acute phase and developing effective therapies are urgently needed. Scutellarin had been found to be beneficial to ischemic injuries and has been shown to have potent effects in clinical application on both stroke and myocardial infarction. However, whether scutellarin improves ischemic brain damage in the acute phase remains unknown. In this study, the protective effects of scutellarin on ischemic brain damage in the acute phase (within 12 h) were illustrated. In middle cerebral artery occlusion and reperfusion (MCAO/R) modeling rats, the Z-Longa score was significantly down-regulated by 25% and 23.1%, and the brain infarct size was reduced by 26.95 ± 0.03% and 25.63 ± 0.02% when responding to high-dose and low-dose scutellarin treatments, respectively. H&E and TUNEL staining results indicated that the neuron loss of the ischemic region was improved under scutellarin treatment. In order to investigate the mechanism of scutellarin’s effects on ischemic brain damage in the acute phase, changes in proteins and metabolites were analyzed. The suppression of scutellarin on the glutamate-inducing excitatory amino acid toxicity was strongly indicated in the study of both proteomics and metabolomics. A molecular docking experiment presented strong interactions between scutellarin and glutamate receptors, which score much higher than those of memantine. Further, by performing a parallel reaction monitoring-mass spectrometry (PRM-MS) study on both the cortex and hippocampus tissue of the ischemic region, we screened the scutellarin-regulating molecules that are involved in both the release and transportation of neurotransmitters. It was found that the aberrant levels of glutamate receptors, including EAAT2, GRIN1, GRIN2B, and GRM1, as well as of other glutamatergic pathway-involving proteins, including CAMKK2, PSD95, and nNOS, were significantly regulated in the ischemic cortex. In the hippocampus, EAAT2, GRIN1, nNOS, and CAM were significantly regulated. Taken together, scutellarin exerts potent effects on ischemic brain damage in the acute phase by regulating the activity of neurotransmitters and reducing the toxicity of excitatory amino acids in in neurons.

Objective To determine the chemical constituents of Paeoniae Radix Rubra and its antipsoriasis activity by ultra performance liquid chromatography-quadrupole-exactive orbitrap mass spectrometry（UHPLC-Q-Exactive-Orbitrap-MS）. Methods Chromatographic conditions were as follows: an ACQUITY UPLC HSS T3 column（2.1 mm×100 mm, 1.8 μm） was used for the gradient elution with 0.1% formic acid water（A）-0.1% formic acid acetonitrile（B） as the mobile phases.Mass spectrometry condition: thermal spray ion source（HESI） was applied, the chromatographic effluent was collected and detected under the negative ion mode with the detection range of m/z 100 ～ 1200. The over proliferation of human immortalized keratinocytes（HaCaT cells） was induced by tumor necrosis factor（TNF）-α to establish a psoriasis cell model. The levels of interleukin-2（IL-2）, interleukin-6（IL-6） and interferon-γ（IFN-γ） in the supernatant of cell culture of each group were determined by ELISA. The effect of Paeoniae Radix Rubra ethanol extraction, paeoniflorin as well as albiflorin in total paeony glycoside（TPG） on psoriasis were determined. Results Totally 61 compounds were identified, including 24 monoterpenoids and their glycosides, 13 flavones and their glycosides, 14 phenolic acids, and 10 tannins. In the cell experiment, compared with the blank group, the levels of IL-2, IL-6 and IFN-γ in the model group were significantly increased（P ＜ 0.01）. Compared with the model group, the levels of IL-2, IL-6 and IFN-γ in the ethanol extract of Paeoniae Radix Rubra, paeoniflorin and albiflorin groups were significantly decreased（P ＜ 0.01）.Conclusion Paeoniae Radix Rubra can be identified in a easy, quick, and accurate way with UHPLCQ-Exactive-Orbitrap-MS technology. The ethanol extract of Paeoniae Radix Rubra, paeoniflorin and albiflorin exerts its anti-psoriasis effect by reducing the levels of IL-2, IL-6, IFN-γ psoriasis related inflammatory factors. This study provides reference for the clinical application, pharmacodynamic substance base study and quality evaluation of Paeoniae Radix Rubra.

Anxiety disorder is a chronic and disabling psychiatric disorder that is more prevalent in females than in males. 11‐Ethoxyviburtinal is an iridoid extracted from Valeriana jatamansi Jones , which has anxiolytic potential. The aim of the present work was to study the anxiolytic efficacy and mechanism of 11‐ethoxyviburtinal in gender‐specific mice. We first evaluated the anxiolytic‐like efficacy of 11‐ethoxyviburtinal in chronic restraint stress (CRS) mice of different sexes through behavioral experiments and biochemical indexes. In addition, network pharmacology and molecular docking were used to predict potential targets and important pathways for the treatment of anxiety disorder with 11‐ethoxyviburtinal. Finally, the influence of 11‐ethoxyviburtinal on phosphoinositide‐3‐kinase (PI3K)/protein kinase B (Akt) signaling pathway, estrogen receptor β (ERβ) expression, and anxiety‐like behavior in mice was verified by western blotting, immunohistochemistry staining, antagonist intervention methods, and behavioral experiments. 11‐ethoxyviburtinal alleviated the anxiety‐like behaviors induced by CRS and inhibited neurotransmitter dysregulation and HPA axis hyperactivity. It inhibited the abnormal activation of the PI3K/Akt signaling pathway, modulated estrogen production, and promoted ERβ expression in mice. In addition, the female mice may be more sensitive to the pharmacological effects of 11‐ethoxyviburtinal. 11‐ethoxyviburtinal may exert its anxiolytic‐like effects through PI3K/Akt and E2/ERβ signaling pathways. Meanwhile, by comparing the male and female mice, gender differences may affect the therapy and development of anxiety disorder.

Abstract Context Levodopa combined with traditional Chinese medicine has a synergistic effect on Parkinson’s disease (PD). Recently, we demonstrated that Nardostachys jatamansi (D. Don) DC. [syn. Patrinia jatamansi D.Don, N. grandiflora DC.] (Valerianaceae) (NJ) can alleviate PD. Objective To explore the synergistic effect of NJ combined with levodopa against PD. Materials and methods The PD model was established by injecting rotenone. Eighty-four Sprague-Dawley rats were randomly divided into seven groups: sham, model, different doses of NJ (0.31, 0.62, or 1.24 g/kg) combined with levodopa (25 mg/kg), and levodopa alone (25 and 50 mg/kg) groups. The synergistic effect of the combination was investigated by pharmacodynamic investigation and detection of expression of nuclear factor erythro2-related factor 2 (Nrf2) and NLR family proteins containing Pyrin-related domain 3 (NLRP3) pathways. Results Compared with the model group, NJ + levodopa (1.24 g/kg + 25 mg/kg) increased the moving distance of PD rats in the open field (2395.34 ± 668.73 vs. 1501.41 ± 870.23, p < 0.01), enhanced the stay time on the rotating rod (84.86 ± 18.15 vs. 71.36 ± 17.53, p < 0.01) and the combination was superior to other treatments. The synergistic effects were related to NJ + levodopa (1.24 g/kg + 25 mg/kg) increasing the neurotransmitter levels by 38.80%-88.67% in PD rats, and inhibiting oxidative stress and NLRP3 pathway by activating Nrf2 pathway. Discussion and conclusions NJ combined with levodopa is a promising therapeutic candidate for PD, which provides a scientific basis for the subsequent clinical combination therapy of levodopa to enhance the anti-PD effect.

Suanzaoren-Wuweizi herb-pair (SWHP), composed of Zizyphi Spinosi Semen (Suanzaoren in Chinese) and Schisandrae Chinensis Fructus (Wuweizi in Chinese), is a traditional herbal formula that has been extensively used for the treatment of insomnia. The study aimed to explore the targets and signal pathways of Suanzaoren-Wuweizi (S-W) in the treatment of anxiety by network pharmacology, and to verify the pharmacodynamics and key targets of SWHP in mice. The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) as well as literature mining were used to obtain the main chemical ingredients of Suanzaoren and Wuweizi. The SwissTargetPrediction platform was used to predict drug-related targets. The GeneCards, TTD, DisGeNET and OMIM databases were used to obtain potential targets for the treatment of anxiety with the chemical components of S-W. Drug-disease intersection genes were selected, and a protein-protein interaction (PPI) network was constructed using STRING. The core targets of S-W in the treatment of anxiety were selected according to the topological parameters, and GO functional enrichment as well as KEGG pathways enrichment analyses were performed for potential targets. The relationship network of the “drug-active ingredient-disease-target-pathway” was constructed through Cytoscape 3.8.0. The pharmacodynamics of SWHP in the treatment of anxiety was evaluated by the elevated plus maze (EPM), the light/dark box test (LDB) and the open field test (OFT). The mechanisms were examined by measuring monoamine neurotransmitters in brain of mice. The results showed that there were 13 active ingredients for the treatment of anxiety in the network. This includes sanjoinenine, swertisin, daucosterol, schizandrer B, wuweizisu C and gomisin-A. Additionally, there were 148 targets, such as AKT1, TNF, SLC6A4, SLC6A3, EGFR, ESR1, HSP90AA1, CCND1, and DRD2, mainly involved in neuroactive ligand-receptor interactions, the Serotonergic synapse pathway and the cAMP signaling pathway. After 1 week of treatment, SWHP (2 and 3 g/kg) induced a significant increase on the percentage of entries into and time spent on the open arms of the EPM. In the LDB test, SWHP exerted anxiolytic-like effect at 2 g/kg. In the open-field test, SWHP (2 g/kg) increased the number of central entries and time spent in central areas. The levels of brain monoamines (5-HT and DA) and their metabolites (5-HIAA, DOPAC) were decreased after SWHP treatment. The anti-anxiety effect of SWHP may be mediated by regulating 5-HT, DA and other signaling pathways. These findings demonstrated that SWHP produced an anxiolytic-like effect and the mechanism of action involves the serotonergic and dopaminergic systems, although underlying mechanism remains to be further elucidated.

目的:建立不同产地野生与栽培赤芍的高效液相色谱(HPLC)指纹图谱并结合灰色关联-逼近理想解排序法(TOPSIS)分析方法评价其质量.方法:采用Agilent ZORBAX SB C18(4.6 mm×250 mm,5 μm)色谱柱,以乙腈-0.1％磷酸水溶液为流动相,梯度洗脱,检测波长254 nm.结果:建立30 批不同产地野生与栽培赤芍的指纹图谱,相似度为0.821～0.989;共标定了15 个共有峰,指认了没食子酸、羟基芍药苷、儿茶素、芍药内酯苷、芍药苷、没食子酸乙酯、没食子酰芍药苷、苯甲酸、1,2,3,4,6-五没食子酰葡萄糖、芍药新苷、苯甲酰芍药苷11 种成分;热图聚类分析显示30 批不同产地野生与栽培赤芍聚为3 类;灰色关联度TOPSIS 分析不同产地野生与栽培赤芍的质量,综合评价河北野生赤芍质量最优;河北栽培、四川栽培赤芍品质较优;黑龙江野生、内蒙古野生、吉林栽培、山东栽培、安徽栽培赤芍质量次之;黑龙江栽培、内蒙古栽培赤芍质量较差.提示河北、四川等区域可作为赤芍的适宜栽培产地.结论:该结果可为赤芍药材的质量评价提供参考,同时也为赤芍栽培品的市场流通及种植选址提供依据.