Quality difference of wild Lysimachiae Herba in the source areas of Sichuan, Chongqing and Guizhou / 国际中医中药杂志

Objective:To determine the contents of quercetin, kaempferol, total flavonoids and extracts in 52 samples of Lysimachiae Herba collected from different origins; To analyze the quality differences of Lysimachiae Herba among different producing areas. Methods:The quercetin and kaempferol contents of the Lysimachiae Herba from Guizhou Province, Sichuan Province and Chongqing were determined by HPLC, and the total flavonoids were determined by Symergy HTX microplate reader. Results:The total content of quercetin and kaempferol in 52 samples was among 0.146 2-2.517 0 mg/g, with an average content of 0.872 6 mg/g, among which the average content of Sichuan was 1.073 2 mg/g, that of Guizhou was 0.705 4 mg/g, and that of Chongqing was 0.865 1 mg/g. Among them, 20 samples reached the standard of the Chinese Pharmacopoeia. The average content of the samples that met the standard was 1.439 7 mg/g. The compliance rate of samples collected in Guizhou, Sichuan and Chongqing reached 12.5%, 62.5%, and 38.8% respectively. The total flavonoid content of 52 samples was among 0.994 2- 3.866 4 mg/g, and 52 samples were in conformity with the ethanol hot extract standard of the Chinese Pharmacopoeia. Conclusions:The total contents of quercetin and kaempferol from different sources in Sichuan, Guizhou and Chongqing are quite different, and the total contents of quercetin and kaempferol collected from the same district and county are also quite different, and the compliance rate is low. There are great differences in total flavonoids in different producing areas and different populations of Lysimachiae Herba samples collected in the field.

Effect of kaempferol on the proliferation, migration and invasion of cervical cancer cells through circFBXW7 / 国际生物医学工程杂志

Objective:To investigate the effect of kaempferol on proliferation, migration, and invasion of cervical cancer SiHa cells by regulating circFBXW7.Methods:SiHa cells were treated with kaempferol at low, medium, and high doses (15, 30, and 60 μmol/L) for 24 h. Untreated SiHa cells were used as the control group. CCK-8 was used to detect the effect of kaempferol on the proliferation of SiHa cells. Transwell was used to detect the effect of kaempferol on the migration and invasion of SiHa cells. Real-time fluorescence quantitative polymerase chain reaction was used to detect the expression of circFBXW7 in SiHa cells. pcDNA and pcDNA-circFBXW7 were transfected into SiHa cells, respectively, and si-NC and si-circFBXW7 were transfected into SiHa cells after adding 60 μmol/L kaempferol treatment for 24 h. The effects of circFBXW7 and its knockdown, circFBXW7, on the proliferation, migration, and invasion of SiHa cells were investigated, and the effects of E-cadherin and N-cadherin protein expression levels were detected by Western Blot. Results:Compared with the control group, the cell value-added, migration, and invasion abilities of the low, medium, and high dose groups were decreased (all P < 0.05) and were dose-related, and the expression of circFBXW7 was increased ( P < 0.05). After transfection with pcDNA-circFBXW7, the expression of circFBXW7 increased ( P < 0.05), while promoting the proliferation, cell migration, and invasion of kaempferol on SiHa cells (all P < 0.05). After transfection with si-circFBXW7, the expression of circFBXW7 decreased ( P < 0.05), while inhibiting the proliferation, cell migration, and invasion of kaempferol on SiHa cells (all P < 0.05). That indicated that the transfection of si-circFBXW7 could attenuate the inhibitory effects of kaempferol on the above oncogenic phenotypes of SiHa cells. Compared with the control group, E-cadherin expression was upregulated, and N-cadherin expression was downregulated in the low, medium, and high dose groups (all P < 0.05) in a dose-related manner. After transfection of pcDNA-circFBXW7 with SiHa cells, the expression of E-cadherin was increased, and the expression of N-cadherin was decreased (all P < 0.05). After transfection of si-circFBXW7 with SiHa cells, the expression of E-cadherin decreased, and the expression of N-cadherin increased (all P < 0.05). Conclusions:Kaempferol can reduce the proliferation, migration, and invasion abilities of SiHa cells by promoting circFBXW7 expression.

Junctional adhesion molecule-like protein as a novel target for kaempferol to ameliorate lung adenocarcinoma / 中西医结合学报

OBJECTIVE@#Although there have been improvements in targeted therapy and immunotherapy, the majority of lung adenocarcinoma (LUAD) patients still lack effective therapies. Consequently, it is urgent to screen for new diagnosis biomarkers and pharmacological targets. Junctional adhesion molecule-like protein (JAML) was considered to be an oncogenic protein and may be a novel therapeutic target in LUAD. Kaempferol is a natural flavonoid that exhibits antitumor activities in LUAD. However, the effect of kaempferol on JAML is still unknown.@*METHODS@#Small interfering RNA was used to knockdown JAML expression. The cell viability was determined using the cell counting kit-8 assay. The proliferation of LUAD cells was evaluated using the 5-ethynyl-2'-deoxyuridine incorporation assay. The migration and invasion of LUAD cells were evaluated by transwell assays. Molecular mechanisms were explored by Western blotting.@*RESULTS@#JAML knockdown suppressed proliferation, migration and invasion of LUAD cells, and JAML deficiency restrained epithelial-mesenchymal transition (EMT) via inactivating the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR) pathway. Using a PI3K activator (740Y-P), rescue experiments showed that phenotypes to JAML knockdown in LUAD cells were dependent on the PI3K/AKT/mTOR pathway. Kaempferol also inhibited proliferation, migration and invasion of A549 and H1299 cells and partially suppressed EMT through the PI3K/AKT/mTOR pathway. Knockdown of JAML ameliorated the inhibitory effect of kaempferol on LUAD cells. Kaempferol exerted anticancer effects by targeting JAML.@*CONCLUSION@#JAML is a novel target for kaempferol against LUAD cells. Please cite this article as: Wu Q, Wang YB, Che XW, Wang H, Wang W. Junctional adhesion molecule-like protein as a novel target for kaempferol to ameliorate lung adenocarcinoma. J Integr Med. 2023; 21(3): 268-276.

Inhibitory Effect of Kaempferol on Proliferation of KG1a Cells and Its Mechanism / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of kaempferol on proliferation of acute myeloid leukemia (AML) KG1a cells and its mechanism.@*METHODS@#Human AML KG1a cells in logarithmic growth stage were taken and set at 25, 50, 75 and 100 μg/ml kaempferol group, another normal control group (complete medium without drug) and solvent control group (add dimethyl sulfoxide) were also set. After 24 and 48 hours of intervention, the cell proliferation rate was detected by CCK-8 assay. In addition, interleukin-6 (IL-6) combined with kaempferol group (Plus 20 μg/l IL-6 and 75 μg/ml kaempferol) was set up, 48 hours after culture, the cell cycle and apoptosis of KG1a cells were detected by flow cytometry, the mitochondrial membrane potential (MMP) of KG1a cells was detected by MMP detection kit (JC-1 method), and the expression of Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) pathway related proteins in KG1a cells were detected by Western blot.@*RESULTS@#The cell proliferation rate of 25, 50, 75 and 100 μg/ml kaempferol group decreased significantly (P<0.05), and with the increase of kaempferol dose (r24 h=-0.990, r48 h= -0.999), the cell proliferation rate decreased gradually (P<0.05). The inhibitory effect of 75 μg/ml kaempferol on cell proliferation reached half of effective dose after 48 hours of intervention. Compared with normal control group, the G0/G1 phase cell proportion and apoptosis rate of cells in 25, 50 and 75 μg/ml kaempferol group increased, while the S phase cell proportion, MMP, phosphorylated JAK2 (p-JAK2)/JAK2 and phosphorylated STAT3 (p-STAT3)/STAT3 protein expression decreased in a dose-dependent manner (r=0.998, 0.994, -0.996, -0.981, -0.997, -0.930). Compared with 75 μg/ml kaempferol group, the G0/G1 phase cell proportion and apoptosis rate of cells in IL-6 combined with kaempferol group decreased, while the S phase cell proportion, MMP, p-JAK2/JAK2 and p-STAT3/STAT3 protein expression increased significantly (P<0.05).@*CONCLUSION@#Kaempferol can inhibit KG1a cell proliferation and induce KG1a cell apoptosis, its mechanism may be related to the inhibition of JAK2/STAT3 signal pathway.

Estudo do potencial antiviral sobre Sars Cov-2 dos compostos identificados nas folhas do extrato bruto de Eugenia pyriformis / Study of the antiviral potential on Sars Cov-2 of compounds identified in the leaves of the gross extract of Eugenia pyriformis / Estudio del potencial antiviral sobre Sars Cov-2 de los compuestos identificados en las hojas del extracto crudo de Eugenia pyriformis

Eugenia pyriformis Cambess (Myrtaceae), conhecida popularmente como uvaia. Em seus frutos são encontrados compostos fenólicos com ação antioxidante e nas folhas foram detectados altos teores de flavonoides e taninos hidrolisados que se mostraram inibidor da protease de 2019 - nCoV e SARS-CoV. Neste sentido, o objetivo deste estudo foi a obtenção do extrato bruto das folhas, a análise da composição química e a possibilidade da ação antiviral frente ao SARS COV-2. O extrato bruto (EB) foi obtido a partir das folhas secas de E. pyriformis, pela técnica de maceração dinâmica com esgotamento do solvente (etanol 90º GL) e concentrado em evaporador rotativo. Seis gramas do EB foram fracionados em cromatografia em coluna, e eluído com hexano, diclorometano, acetato de etila e metanol, as frações foram concentradas em um evaporador rotativo (Tecnal TE-210). O EB e as frações foram identificadas por cromatografia líquida de alta eficiência à espectrometria de massas de alta resolução (CLAE-ESI/qTOF). A identificação química do extrato bruto e frações das folhas de E. pyriformis evidenciou a presença de compostos fenólicos destacando os ácidos fenólicos, flavonoides e taninos. De forma complementar, foi realizado um levantamento bibliográfico sobre a provável ação antiviral dos compostos fenólicos e taninos presentes nas folhas de uvaia. Os resultados evidenciaram que os flavonoides quercetina e kaempferol possuem ação antiviral quando se ligam a glicoproteína do envelope ou capsídeo viral interferindo na ligação e penetração do vírus na célula. Este resultado coloca as folhas de E. pyriformis na lista de plantas com ação antiviral.

Eugenia pyriformis Cambess (Myrtaceae), popularly known as uvaia. In its fruits, phenolic compounds with antioxidant action are found and in the leaves, high levels of flavonoids and hydrolyzed tannins were detected, which proved to be an inhibitor of the 2019 protease - nCoV and SARS-CoV. In this sense, the objective of this study was to obtain the crude extract of the leaves, the analysis of the chemical composition and the possibility of antiviral action against SARS COV-2. The crude extract (EB) was obtained from the dried leaves of E. pyriformis, by the dynamic maceration technique with solvent exhaustion (ethanol 90º GL) and concentrated in a rotary evaporator. Six grams of EB were fractionated in column chromatography, and eluted with hexane, dichloromethane, ethyl acetate and methanol, the fractions were concentrated on a rotary evaporator (Tecnal TE-210). EB and fractions were identified by high performance liquid chromatography using high resolution mass spectrometry (HPLC-ESI/qTOF). The chemical identification of the crude extract and fractions of E. pyriformis leaves evidenced the presence of phenolic compounds, highlighting phenolic acids, flavonoids and tannins. In addition, a bibliographic survey was carried out on the probable antiviral action of phenolic compounds and tannins present in uvaia leaves. The results showed that the flavonoids quercetin and kaempferol have antiviral action when they bind to the envelope glycoprotein or viral capsid, interfering with the binding and penetration of the virus into the cell. This result places E. pyriformis leaves in the list of plants with antiviral action.

Eugenia pyriformis Cambess (Myrtaceae), conocida popularmente como uvaia. En sus frutos se encuentran compuestos fenólicos con acción antioxidante y en las hojas se detectaron altos contenidos de flavonoides y taninos hidrolizados que demostraron inhibir la proteasa de 2019 - nCoV y SARS-CoV. En este sentido, el objetivo de este estudio fue obtener el extracto crudo de las hojas, el análisis de la composición química y la posibilidad de acción antiviral contra el SARS COV-2. El extracto crudo (EB) se obtuvo a partir de las hojas secas de E. pyriformis, mediante la técnica de maceración dinámica con agotamiento del disolvente (etanol 90º GL) y se concentró en evaporador rotatorio. Seis gramos de EB se fraccionaron en cromatografía en columna, y se eluyeron con hexano, diclorometano, acetato de etilo y metanol, las fracciones se concentraron en un evaporador rotatorio (Tecnal TE-210). El EB y las fracciones se identificaron mediante cromatografía líquida de alta resolución a espectrometría de masas de alta resolución (HPLC-ESI/qTOF). La identificación química del extracto crudo y de las fracciones de las hojas de E. pyriformis mostró la presencia de compuestos fenólicos destacando los ácidos fenólicos, los flavonoides y los taninos. De forma complementaria, se realizó un estudio bibliográfico sobre la probable acción antiviral de los compuestos fenólicos y los taninos presentes en las hojas de la uva. Los resultados mostraron que los flavonoides quercetina y kaempferol tienen acción antiviral cuando se unen a la glicoproteína de la envoltura o cápside viral, interfiriendo en la unión y penetración del virus en la célula. Este resultado sitúa a las hojas de E. pyriformis en la lista de plantas con acción antiviral.

Medicinal plants with antimalarial activities mediated via glycogen synthase kinase-3 beta (GSK3β) inhibition

@#Many of the therapeutic effects of plant extracts and bioactive compounds appear related to their immunomodulatory effects and impact on the host immune system. The immune response is desirable to mitigate established infections and, in the case of severe malaria, is a feasible approach to dealing with the overwhelming cytokine response. Glycogen synthase kinase-3 (GSK3), a Ser/Thr kinase that is a central regulator of the cytokine response, is a promising antimalarial drug target. In this review, we discussed our ongoing research projects, which include assessing the antimalarial activities of medicinal plants and their bioactive compounds, immunomodulatory activities mediated by GSK3, and the potential inflammatory pathway involved in malarial infection.

Study on protective mechanism of kaempferol against cerebral ischemia/reperfusion injury in vitro and in vivo / 中国药房

OBJECTIVE To explore the protective mechanism of kaempferol （KAE）on cerebral ischemia/reperfusion （CI/R） injury in vitro and in vivo . METHODS Firstly，the potential targets of KAE in the treatment of CI/R injury were preliminarily screened by network pharmacology. AutoDock Vina software was used to conduct molecular docking between KAE and the top 10 core targets ，and the binding affinity was used as the evaluation standard to further clarify the possible mechanism of KAE in treating CI/R injury. Finally ，the above results were verified by in vivo and in vitro experiments. The oxygen glucose deprivation/ reperfusion（OGD/R）HT22 cell injury model and the middle cerebral artery occlusion （MCAO）rat model were constructed. The cell activity was detected by CCK- 8 method. The neural function score and TTC staining were performed on the rats and their brain tissues. The phosphorylation levels of protein kinase B （Akt）and Src in HT 22 cells and brain tissue of rats were detected by Western blot. RESULTS The results of network pharmacology screening showed that KAE in the treatment of CI/R injury was closely related to 10 core targets including prostaglandin-endoperoxide synthase 2，matrix metalloproteinase 9，JUN，Akt1，tumor necrosis factor ，caspase-3，mitogen activated protein kinase 8，intercellular cell adhesion molecule 1，vascular cell adhesion molecule 1 and Src. The results of molecular docking showed that KAE was stably bound with Akt 1 and Src . The results of in vitro and in vivo experiments showed that KAE could significantly improve the survival rate of OGD/R-injuried HT 22 cells （P＜0.05）， significantly reduced the neurological function score of MCAO model rats （P＜0.05），significantly reduces the volume of cerebral infarction in rats （P＜0.05），and significantly increased the phosphorylation levels of Akt and Src in HT 22 cells and brain tissue of rats（P＜0.05），which showed a dose dependent trend. CONCLUSIONS KAE may play a neuroprotective role by regulating the phosphorylation expression of Akt and Src ，thus treating CI/R injury.

Comparative characterization of cinnamon, cinnamaldehyde and kaempferol for phytochemical, antioxidant and pharmacological properties using acetaminophen-induced oxidative stress mouse model / Caracterización comparativa de canela, cinamaldehído y kaempferol para propiedades fitoquímicas, antioxidantes y farmacológicas utilizando el modelo de ratón con estrés oxidativo inducido por acetaminofeno

This study was aimed to explore the comparative efficacy of cinnamon bark extract, cinnamaldehyde and kaempferol against acetaminophen (APAP)-induced oxidative stress. Cinnamon bark extract, cinnamaldehyde and kaempferol were utilized or in-vivo analysis. From the results of in-vitro screening tests, cinnamon ethanolic extract was selected for in-vivo study in mouse model. For this, Balb/c albino mice were treated with cinnamon ethanolic extract (200 mg/kg), cinnamaldehyde (10 mg/kg) and kaempferol (10 mg/kg) orally for 14 days followed by single intraperitoneal administration of APAP during 8 hours. Blood and organ samples were collected for biochemical and histopathological analysis. The results showed that cinnamon bark ethanolic extract, cinnamaldehyde and kaempferol ameliorated APAP-induced oxidative stress and organ toxicity in mice. In conclusion, cinnamaldehyde and kaempferol possess comparable antioxidant potential even at 20-times less dose as compared to cinnamon bark ethanolic extract suggesting therapeutic potential in oxidative stress-related disorders.

Este estudio tuvo como objetivo explorar la eficacia comparativa del extracto de corteza de canela, cinamaldehído y kaempferol contra el estrés oxidativo inducido por acetaminofén (APAP). Se utilizaron extracto de corteza de canela, cinamaldehído y kaempferol para el análisis in vivo. De los resultados de las pruebas de detección in vitro, se seleccionó el extracto etanólico de canela para estudio in vivo en modelo de ratón. Para ello, los ratones albinos Balb/c fueron tratados con extracto etanólico de canela (200 mg/kg), cinamaldehído (10 mg/kg) y kaempferol (10 mg/kg) por vía oral durante 14 días, seguido de la administración intraperitoneal única de APAP durante 8 horas. Se recogieron muestras de sangre y órganos para análisis bioquímicos e histopatológicos. Los resultados mostraron que el extracto etanólico de la corteza de canela, el cinamaldehído y el kaempferol mejoraron el estrés oxidativo inducido por APAP y la toxicidad orgánica en ratones. En conclusión, el cinamaldehído y el kaempferol poseen un potencial antioxidante comparable, incluso a una dosis 20 veces menor en comparación con el extracto etanólico de la corteza de canela, lo que sugiere un potencial terapéutico en los trastornos relacionados con el estrés oxidativo.

HPLC determination of the content for six constituents before and after salt processing of Plantaginis Semen from different habitats / 国际中医中药杂志

Objective:To compare the content change of 6 constituents in Plantaginis Semen from different habitats before and after salt processing. Methods:HPLC method was used to quantitatively analyze 6 ingredients in Plantaginis Semen and processed with salt including geniposidic acid, plantagoguanidinic acid, quercetin, kaempferol, verbascoside and isoverbascoside. Results:The geniposidic acid, plantagoguanidinic acid, quercetin, kaempferol, verbascoside and isoverbascoside were well separated. The linear ranges of which were 0.259 2-3.628 8 μg ( r=0.999 8), 0.054 3-0.760 5 μg ( r=0.999 6), 0.030 0-0.420 6 μg ( r=0.999 4), 0.055 6- 0.777 8 μg ( r=0.999 5), 0.287 0-4.018 0 μg ( r=0.999 8), 0.033 1-0.463 1 μg ( r=0.999 7), respectively. Average recovery rates were 98.68%, 98.46%, 98.87%, 98.99%, 98.34%, 98.75% ( n=6), respectively. There were mild differences in the contents of 6 ingredients of 8 batches of Plantaginis Semen from 5 different habitats. There were no obvious differences between the raw products and the products after salt process in Plantaginis Semen. The content of flavonoids, geniposidic acid and isoverbascoside in Plantaginis Semen were significantly increased after salt process, while the content of verbascoside was reduced. Conclusion:HPLC method to quantitatively analyze the 6 constituents in Plantaginis Semen before and after salt process could provided a reference for the quality change and the material basis for the efficacy of Plantaginis Semen before and after salt process.

An Exploration on Mechanisms of Treating Different Diseases with Same Method of Qinggan Huayu Granules in Treating NAFLD and Liver Cancer Based on Network Pharmacology / 中国实验方剂学杂志

Objective:To reveal the effective components， targets and possible mechanisms of Qinggan Huayu granules in the treatment of non-alcoholic fatty liver disease （NAFLD） and liver cancer based on network pharmacology and experimental verification， and to provide a basis for its rational interpretation of treating different diseases with same method for NAFLD and liver cancer. Method:Based on databases of traditional Chinese medicine and disease， the network pharmacology was used to screen main active compounds and potential targets of Qinggan Huayu granules for NAFLD and liver cancer. STRING 11.0 was used to analyze the interaction between potential targets. The core targets were selected from the interaction targets by cytoHubba plug-in. The gene ontology （GO） function and the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis were performed on the target by Metascape database. At the same time， <italic>in vitro</italic> experiments were conducted to validate the effect of kaempferol， one of the main active ingredients of Qinggan Huayu granules， on hepatocellular carcinoma cell model and NAFLD cell model. Result:A total of 43 potential targets of Qinggan Huayu granules for for NAFLD and liver cancer were screened， corresponding to 136 active ingredients in 8 herbal medicines. Through enrichment analysis of potential targets， there were 20 biological processes， 13 molecular functions， 9 cellular components and 15 signaling pathways. Qinggan Huayu granules regulated biological behaviors of tumors related to liver cancer and NAFLD （such as apoptosis inhibition and oxidative stress） mainly through kaempferol， quercetin， luteolin and other active ingredients for Caspase-3 （CASP3）， tumor protein p53 （TP53）， vascular endothelial growth factor A （VEGFA） and other hub genes. <italic>In vitro</italic> experiments revealed that kaempferol could inhibit cell proliferation in a dose-dependent manner in hepatocellular carcinoma cell model. And kaempferol could modulate the levels of malondialdehyde （MDA） and glutathione peroxidase （GPx）， which were the molecular markers of oxidative stress of NAFLD cell model. Kaempfero also regulated the expression level of CASP3 in hepatocellular carcinoma cell model and NAFLD cell model. Conclusion:The main mechanism of Qinggan Huayu granules in treating liver cancer and NAFLD with concept of treating different diseases with same method is related to systematic synergy effect of multiple compounds （represented by quercetin， luteolin and kaempferol）， multiple targets （represented by VEGFA， TP53 and CASP3） and multiple signaling pathways （represented by oxidative stress and cell apoptosis）.

Anti-tumor Effect and Mechanism of Kaempferol： A Review / 中国实验方剂学杂志

Cancer is a threat to human health. New treatments for cancer can significantly prolong the survival time of patients， but fail to improve the adverse reactions induced by chemoradiotherapy. Improving patient outcomes still requires the effort of cancer researchers. In recent years， the anti-tumor effects of active components from Chinese herbs have received wide attention. Kaempferol， a flavonoid mainly found in the medicinal plant Kaempferia galanga， can be used to treat obesity， cardiovascular diseases， diabetes and other diseases. It has also exhibited good efficacy in inhibiting the occurrence and development of liver cancer， colon cancer， lung cancer， ovarian cancer， and other malignant tumors. Kaempferol mainly exerts the anti-cancer effect by inducing apoptosis. Specifically， it promotes the production of intracellular reactive oxygen species （ROS） and triggers cell apoptosis through the mitochondrial pathway. Besides， it is capable of interfering with the cancer cell cycle， causing most cancer cells to arrest in the G2/M phase， and inducing cell autophagy， a programmed cell death， thus inhibiting cell migration and invasion and angiogenesis， synergistically improving chemotherapeutic drug efficacy， and reducing adverse effects. Kaempferol acts on a series of intracellular and extracellular targets to participate in the regulation of tumor cell signaling pathways， involving phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）， epidermal growth factor receptor （EGFR）， mitogen activated protein kinase （MAPK）， and Wnt signaling pathways， with the PI3K/Akt signaling pathway being most significant. In addition， kaempferol also plays an important regulatory role in tumor epigenetics. This paper reviewed the anti-tumor effect and mechanism of kaempferol， aiming to provide reference for in-depth study on its prevention and treatment of tumor and the development of new anti-tumor drugs.

Isomeric discrimination of kaempferol versus luteolin by online energy-resolved mass spectrometry / 中国中药杂志

Isomers are widely distributed in Chinese herbal medicines,and can be discriminated by energy-resolved mass spectrometry( ER-MS). However,ER-MS was performed through direct injection of reference compounds with syringe pump,which encountered a significant technical barrier for high-throughput and automated measurements. Herein,online ER-MS was conducted using LC-MS platform,and a pair of isomers,kaempferol vs luteolin,were employed as a case study to illustrate and assess the utility of online ER-MS for isomeric discrimination. High-resolution tandem mass spectrometry data of both flavonoids were acquired on LC-QE-Orbitrap-MS,and the fragmentation pathways responsible for the primary fragment ions were proposed. The primary signal in MS1 occurred at m/z 285( [M-H]-),and the primary signals of either compound generated by retro-Diels-Alder fragmentation were observed at m/z 151 and 133. The spectral information was subsequently transferred onto LC-Qtrap-MS platform to carry out online ER-MS. Two precursor-to-product ion transition candidates were constructed as m/z 285>151 and 285>133,and either afterward derived a set of pseudo-ion transitions( PITs) and so forth,exactly corresponding to a series of progressive collision energies( eg-5,-8,-11 e V,and so on). All PITs were typed into the monitoring list of multiple reaction monitoring program to generate the peak area datasets. Either dataset was normalized using the highest values in the set and imported into Graph Pad Prism software to plot the Gaus-sian-shaped curve that was termed as the break-down graph. The apex of the regressive curve was termed as optimal collision energy( OCE). The OCE values corresponding to m/z 285>151 were calculated as-29. 06 e V and-35. 71 e V for kaempferol and luteolin,respectively. In the case of m/z 285>133,the OCEs were yielded as-44. 15 e V for kaempferol and-49. 01 e V for luteolin. With re-ference to their chemical structures,the location of hydroxyl group was regarded to be responsible for the differences of either m/z 285>151 or 285>133 between the isomers,attributing to their different bond properties. Above all,online ER-MS offers an eligible tool for isomeric discrimination,and provides meaningful information for the accurate chemical composition characterization based on LC-MS,which is not limited to Chinese herbal medicines.

Prediction of the pharmacological characteristics and therapeutic mechanism of Tripterygium wilfordii in treating oral lichen planus based on network pharmacology / 口腔疾病防治

Objective@#To explore the potential mechanism of the main active component Tripterygium wilfordii in the treatment of oral lichen planus based on network pharmacology.@*Methods@#The components of Tripterygium wilfordii and targets were searched through the Traditional Chinese Medicine system pharmacology database and analysis platform (TCMSP) and the Traditional Chinese Medicine integrated database (TCMID) databases. The related targets of oral lichen planus (OLP) were obtained through databases such as Gene Cards. The OLP targets were mapped by Venn analysis to the targets of Tripterygium wilfordii to screen out the common targets as the treatment of OLP targets of Tripterygium wilfordii. The Cytoscape software and STRING were used to construct a chemical component-target network and protein-protein interaction network, a network analyzer was used to compute the network topology properties, a cluster profiler software was used to analyze the GO classification enrichment analysis and KEGG signal path analysis, and a Tripterygium wilfordii chemical components-targets-pathway network diagram was constructed. @*Results@#Twenty-three components and 44 OLP treatmenttargets of Tripterygium wilfordii were obtained. The key active ingredients of Tripterygium wilfordii in the treatment of OLP are triptolide, kaempferol, and tangerine peel. The key targets include TNF and AKT1. The GO classification enrichment analysis obtained 63 GO terms, which are mainly involved in the leukocyte differentiation and reaction to lipopolysaccharides. The KEGG analysis identified 111 signaling pathways, which are mainly related to the TNF signaling pathway and IL17 signaling pathway. @*Conclusion@#Tripterygium wilfordii in the treatment of OLP. This study can provide a theoretical basis for further research to explore drugs with high activity and low toxicity to treat OLP from Tripterygium wilfordii.

Kaempferol promotes osteogenic differentiation of mouse bone marrow mesenchymal cells under tension stress via the mTORC1 signaling pathway / 口腔疾病防治

Objective @#To investigate the activation of the mammalian target of rapamycin complex 1 (mTORC1) signaling pathway molecules during the process by which kaempferol (Kae) promotes osteogenic differentiation of mouse bone marrow mesenchymal cells (BMMCs) under cyclic and uniaxial tension.@*Methods @#BMMCs isolated and cultured in vitro were subjected to uniaxial dynamic tension with a 10% shape variable. The appropriate concentration of Kae was selected by cytotoxicity testing. The endogenous mTOR signal was inhibited by pp242. Four hours after traction, alkaline phosphatase (ALP) and osteocalcin (OCN) were detected by chemical colorimetry and ELISA, and the relative concentration of intracellular calcium was detected by flow cytometry. Phosphorylation of mTOR, 4E/BP1, and ribosomal protein S6 kinases (S6K), which are the main molecules of the endogenous mTORC1 signaling pathway, and expression of osteogenic transcription factors (Runx2 and Osterix) were detected by western blotting (WB), and mRNA expression levels of the above factors were detected by qRT-PCR.@*Results @# The cytotoxicity test showed that 10 μmol/L Kae had little inhibitory effect on cell proliferation but had the strongest osteogenic ability. Four hours after stretching, Kae effectively promoted the osteogenic differentiation of BMMCs. The expression of ALP was （153.04 ± 18.72） U/mg, the expression of OCN was （1.64 ± 0.25） U. The mRNA and protein levels of Runx2 and Osterix were upregulated, and the intracellular calcium content was decreased. The mRNA and protein phosphorylation of mTOR and S6K was upregulated, and the opposite effect was observed with 4E/BP1. After pp242 was added to inhibit mTOR signaling, mTOR and S6K mRNA and protein phosphorylation were downregulated, but 4E/BP1 mRNA and protein phosphorylation was upregulated. The osteogenic differentiation of BMMCs was also significantly inhibited, mRNA and protein expression of Runx2 and Osterix were significantly downregulated, ALP and OCN expression were downregulated, and intracellular calcium content was increased. @* Conclusion@#Kae promotes osteogenic differentiation of mouse BMMCs under uniaxial dynamic tension through the mTORC1 signaling pathway.

Anti-viral and anti-inflammatory effects of kaempferol and quercetin and COVID-2019: A scoping review

Severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) is a novel coronavirus identified at the end of 2019. It is recognized as the causative agent of coronavirus disease 2019 (COVID-19). Flavonoids have been shown to exhibit therapeutical effect on complications related to COVID-19. The present study reviews possible therapeutic benefits of flavonoids on SARS-CoV-2. The Web of Science, PubMed, Scopus, and Google Scholar were searched using keywords: 'COVID-19', 'SARS-CoV-2', 'Kaempferol' and 'Quercetin' in the Title/Abstract. Relevant published articles in the English language until August 2020 were considered. Kaempferol and quercetin showed antiviral properties such as inhibition of protein kinase B and phosphorylation of protein kinase and blocking effects on a selective channel (3a channel) expressed in SARS-CoV infected cells. They also reduced the level of reactive oxygen species, expression of inducible nitric oxide synthase, pro-inflammatory mediators including TNF-α, IL-1α, IL-1β, IL-6, IL-10, and IL-12 p70, and chemokines. Kaempferol and quercetin might exert beneficial effects in the control or treatment of COVID-19 because of their antiviral, antioxidant, anti-inflammatory, and immunomodulatory effects.

Exploration of bioflavonoids targeting dengue virus NS5 RNA-dependent RNA polymerase: In silico molecular docking approach

Dengue viral infection becomes highly epidemic and rashes the economic stability of most of the developing countriesdue to its wide prevalence with limited therapeutic ailments. Alarming demographic data urge the need for thedevelopment of new antiviral agents which are safe and efficacious. This study aimed to evaluate the antiviral potentialof bioflavonoids (apigenin, hesperidin, kaempferol, myricetin, and naringenin) against dengue virus nonstructural(NS)5 RNA-dependent RNA polymerase (RdRp) by AutoDock and tox prediction tools. The results of moleculardocking analysis strongly suggested that the lead phytocomponents such as apigenin, hesperidin, and kaempferolreveal potential RdRp inhibition as ascertained by its interaction with core active amino acid residues (710 SER, 729ARG, and 737 ARG) on the target. Apigenin exhibited the best binding affinity of −8.28kcal/mol with RdRp, followedby kaempferol (−7.00 kcal/mol), myricetin (−4.37 kcal/mol), naringenin (−4.35 kcal/mol), and hesperidin(−3.20 kcal/mol). The present research finding clearly advocates that plant-derived bioflavonoids possess excellent antiviralproperty against the selected target.

Effect of kaempferol on the proliferation, migration, invasion, and apoptosis of human hepatoma Bel-7402 cells / 临床肝胆病杂志

ObjectiveTo investigate the effect of kaempferol on the proliferation, migration, invasion, and apoptosis of human hepatoma Bel-7402 cells and related molecular mechanism. MethodsHepatoma Bel-7402 cells cultured in vitro were randomly divided into control group and low-, middle-, and high-concentration experimental groups. The experimental groups were treated with low-, middle-, and high-concentration kaempferol (25, 50, and 100 μmol/L), and the control group was treated with an equal volume of dimethyl sulfoxide. CCK-8 assay was used to observe the effect of kaempferol on the viability of Bel-7402 cells; plate colony formation assay was used to evaluate the effect of kaempferol on cell colony formation ability; wound healing assay and Transwell chamber were used to observe the effect of kaempferol on cell migration and invasion; Western blot was used to measure the expression of apoptosis- and cycle-related proteins. A one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsAfter 24 hours of treatment, the cell viability was 100.00%±2.72% in the control group and 75.70%±2.42%, 62.79%±2.45%, and 43.41%±2.11%, respectively, in the low-, middle-, and high-concentration experimental groups, and compared with the control group, the experimental groups had a significant reduction in cell viability (all P＜0.05). The number of cell colonies was 923.3±35.2 in the control group and 682.7±24.4, 464.0±22.0, and 327.3±14.0, respectively, in the low-, middle-, and high-concentration experimental groups, and compared with the control group, the experimental groups had a significant reduction in cell colony formation ability (all P＜0.05). After 24 hours of treatment, the relative migration rate was 100.00%±1.11% in the control group and 63.33%±1.16%, 51.72%±3.23%, and 37.18%±2.71%, respectively, in the low-, middle-, and high-concentration experimental groups, and the number of transmembrane cells was 212.0±3.0 in the control group and 134.0±2.0, 71.0±2.0, and 34.0±1.0, respectively, in the low-, middle-, and high-concentration experimental groups; compared with the control group, the experimental groups had significant reductions in relative migration rate and number of transmembrane cells (all P＜0.05). After 48 hours of treatment, compared with the control group, the low-, middle-, and high-concentration experimental groups had a significant reduction in the expression of the anti-apoptotic protein Bcl-2 (all P＜0.05), a significant increase in the expression of the pro-apoptotic protein Bax (all P＜0.05), and a significant reduction in the expression of C＜italic/＞yclinD1 (all P＜005). ConclusionKaempferol can inhibit the proliferation, migration, and invasion of human hepatoma Bel-7402 cells and promote the apoptosis of such cells, possibly by regulating the apoptosis proteins Bax and Bcl-2 and downregulating the expression of CyclinD1.

Non-diterpenoid alkaloids in aerial parts of Aconitum carmichaelii / 中草药

Objective: To study the chemical constituents in the aerial parts of Aconitum carmichaelii. Methods: The air-dried arial parts of A. carmichaelii were powdered and extracted with methanol by percolation extraction. After the removal of solvent under reduced pressure, the crude extract was dissolved in 1.5% HCl solution, and then extracted by ethyl acetate to obtain the total crude extract. The compounds were isolated and purified by column chromatography and identified by spectral analyses (MS, 13C-NMR, and 1H-NMR). Results: Fifteen compounds were isolated from A. carmichaelii and characterized as indol-3-carboxylic acid (1), corchoionol C (2), β-sitosterol-3-O-β-D-glucoside-6’-palmitate (3), (+)-pinoresinol (4), (+)-N-formylnorglaucine (5), oxoglaucidaline (6), glaucine (7), (+)-cataline (8), kaempferol-7-O-α-L-rhamnofuranoside (9), kaempferol-3-O-β-(2″-acetyl)-galactopyranoside (10), megastigmane (11), kaempferol-7-O-α-L-arabinoside (12), kaempferol-3-O-β-D-xylopyranoside (13), kaempferol-3-O-β-D- glucopyranoside (14), and quercetin-3-O-β-D-galactopyranoside (15). Conclusion: All compounds are isolated from aerial parts of A. carmichaelii for the first time, and compounds 1-3,5-6,8-15 are isolated from this plant for the first time.

Chemical constituents from ethyl acetate extract of Eupatorium adenophorum / 中草药

Objective: To study the chemical constituents of the Eupatorium adenophorum. Methods: The chemical constituents were isolated and purified by silica gel chromatography repeatedly from the ethyl acetate extract of E. adenophorum, and their structures were identified by spectral analysis and chemical methods. Results: Fourteen compounds were isolated from E. adenophorum and identified as p-hydroxybenzoic acid ethyl ester (1), (1R,4R)-8aα-hydroxy-1-isopropyl-4,7-dimethyl- 1,2,3,4,6,8a-hexahydro-naphthalene-2,6-dione (2), daedalin A (3), 10-oxo-7-hydroxy-nordehydrotremetone (4), caffeoyl acetate (5), syringic acid (6), 3-hydroxy-4-(1-oxo-ethane)-benzoic acid (7), ferulic acid (8), p-hydroxyphenylethyl alcohol (9), protocatechuic acid ethyl ester (10), 4-hydroxy-3-isopropyl benzoic acid (11), balanophonin (12), indole-3-carboxylic acid (13), and 6-methoxy kaempferol (14). Conclusion: Compound 11 is obtained from natural products for the first time, compounds 1, 3, 6, 7, 9, 10, 12-14 are obtained from this genus for the first time, and compound 5 is obtained from this plant for the first time.

Exploring active compounds of Da-Yuan-Yin in treatment of COVID-19 based on network pharmacology and molecular docking method / 中草药

Objective: To explore the active compound of Da-Yuan-Yin for treatment of coronavirus disease 2019 (COVID-19). Methods: Based on traditional Chinese medicine system pharmacology platform (TCMSP), the chemical composition and targets of Arecae Semen, Magnoliae Officinalis Cortex, Tsaoko Fructus, Anemarrhenae Rhizoma, Paeoniae Radix Alba, Scutellariae Radix, and Glycyrrhizae Radix et Rhizoma were screened. The targets of corresponding gene were searched through UniProt, GeneCards databases, and then Cytoscape3.2.1 was used to build compound-targets (genes) networks. The enrichment of gene ontology (GO) function analysis by DAVID and the pathway enrichment analysis by Kyoto Encyclopedia of Genes and Genomes (KEGG) were carried out, the mechanism of its action was predicted. Results: The compound-target network contained 141 compounds and 267 corresponding targets, and the key targets involved PTGS2, HSP90AA1, ESR1, AR, NOS2, etc. The function enrichment analysis of GO was 522 (P < 0.05), of which there were 421 biological processes (BP) items, and 38 related items of cell composition (CC),and 63 molecular function (MF) items. There were 25 signal pathways (P < 0.05) in the KEGG pathway enrichment screening, involving small cell lung cancer, non-small cell lung cancer and T cell receptor signaling pathways, etc. The results of molecular docking showed that the affinity of quercetin, kaempferol, baicalin and other core compounds was similar to recommended drugs recommended in the treatment of COVID-19. Conclusion: The active compounds in Da-Yuan-Yin may regulate multiple signaling pathways by binding angiotensin converting enzyme II (ACE2) and acting on targets such as PTGS2, HSP90AA1 and ESR1 to inhibit COVID-19.