Mechanism of Key Ingredient of Astragalus membranaceus on Lung Adenocarcinoma via PI3K/AKT Signaling Clarified by Utilizing Network Pharmacology Approach and Experimental Validation / 中国结合医学杂志

OBJECTIVE@#To investigate the mechanism of the effect of Astragalus membranaceus (A. membranaceus) on lung adenocarcinoma at the molecular level to elucidate the specific targets according to the network pharmacology approach.@*METHODS@#The active components of A. membranaceus and their potential targets were collected from the Traditional Chinese Medicine Systems Pharmacology Database. Lung adenocarcinoma-associated genes were acquired based on GeneCards, Online Mendelian Inheritance in Man (OMIM), PharmGKB, and Therapeutic Targets databases. The PI3K/AKT signaling pathway-related genes were obtained using Reactome portal. Networks of "ingredient-target" and "ingredient-target-pathway-disease" were constructed using the Cytoscape3.6.0 software. The relationships among targets were analyzed according protein-protein interaction (PPI) network. Finally, molecular docking was applied to construct the binding conformation between active ingredients and core targets. Cell counting kit 8 (CCK8) and Western blot assays were performed to determine the mechanism of the key ingredient of A. membranaceus.@*RESULTS@#A total of 20 active components and their 329 targets, and 7,501 lung adenocarcinoma-related genes and 130 PI3K/AKT signaling pathway-related genes were obtained. According to Venn diagram and PPI network analysis, 2 mainly active ingredients, including kaempferol and quercetin, and 6 core targets, including TP53, MAPK1, EGF, AKT1, ERBB2, and EGFR, were identified. The two important active ingredients of A. membranaceus, kaempferol and quercetin, exert the therapeutic effect in lung adenocarcinoma partly by acting on the 6 core targets (TP53, MAPK1, EGF, AKT1, ERBB2, and EGFR) of PI3K/AKT signaling pathway. Expressions of potential targets in lung adenocarcinoma and normal samples were analyzed by using UALCAN portal and found that ERBB2 was overexpressed in lung adenocarcinoma tissues and upregulation of it correlated with clinicopathological characteristics. Finally, quercetin repressed viabilities of lung adenocarcinoma cells by targeting ERBB2 on PI3K/AKT signaling confirmed by CCK8 and Western blot.@*CONCLUSION@#Our finding unraveled that an active ingredient of A. membranaceus, quercetin, significantly inhibited the lung adenocarcinoma cells proliferation by repressing ERBB2 level and inactivating the PI3K/AKT signaling pathway.

Progress in Mechanism of Astragalus membranaceus and Its Chemical Constituents on Multiple Sclerosis / 中国结合医学杂志

The primary chemical components of Astragalus membranaceus include polysaccharides, saponins, flavonoids, and amino acids. Recent studies have shown that Astragalus membranaceus has multiple functions, including improving immune function and exerting antioxidative, anti-radiation, anti-tumor, antibacterial, antiviral, and hormone-like effects. Astragalus membranaceus and its extracts are widely used in clinical practice because they have obvious therapeutic effects against various autoimmune diseases and relatively less adverse reaction. Multiple sclerosis (MS) is an autoimmune disease of central nervous system (CNS), which mainly caused by immune disorder that leads to inflammatory demyelination, inflammatory cell infiltration, and axonal degeneration in the CNS. In this review, the authors analyzed the clinical manifestations of MS and experimental autoimmune encephalomyelitis (EAE) and focused on the efficacy of Astragalus membranaceus and its chemical components in the treatment of MS/EAE.

Chemical composition analysis and value evaluation of stems and leaves of Astragalus membranaceus var. mongholicus / 中国中药杂志

This study aimed to provide data support for resource utilization of the stems and leaves of Astragalus membranaceus var. mongholicus(SLAM) by analyzing and evaluating the chemical constituents. The crude protein, crude fiber, and soluble saccharide of SLAM were analyzed by Kjeldahl method, filtration method, and UV-Vis spectrophotometry, respectively. The nucleosides, amino acids, flavonoids, and saponins of SLAM were analyzed by ultraperformance liquid chromatography-triple quadrupole mass spectrometry(UPLC-TQ-MS). Combined with principal component analysis(PCA), the quality difference of resource components of SLAM was comprehensively evaluated. The results showed that the average content of crude protein, crude fiber, total polysaccharide, and redu-cing sugar in SLAM was 5.11%, 30.33%, 11.03 mg·g~(-1), and 31.90 mg·g~(-1), respectively. Six nucleosides, 15 amino acids, 22 flavonoids, and one saponin were detected, with an average content of 1.49 mg·g~(-1), 6.00 mg·g~(-1), 1.86 mg·g~(-1), and 35.67 μg·g~(-1), respectively. The content of various types of chemical components in SLAM differed greatly in different harvesting periods and growing years. The results of PCA showed that the quality of SLAM produced in Ningxia was superior. The results can provide references for the utilization of SLAM.

Protective effect and mechanism of Astragalus membranaceus and Angelica sinensis compatibility against triptolide-induced hepatotoxicity by regulating Keap1/Nrf2/PGC-1α / 中国中药杂志

This paper aims to investigate the protective effect and mechanism of Astragalus membranaceus and Angelica sinensis before and after compatibility against triptolide(TP)-induced hepatotoxicity. The experiment was divided into a blank group, model group, Astragalus membranaceus group, Angelica sinensis group, and compatibility groups with Astragalus membranaceus/Angelica sinensis ratio of 1∶1, 2∶1, and 5∶1. TP-induced hepatotoxicity model was established, and corresponding drug intervention was carried out. The levels of alanine transaminase(ALT), aspartate transaminase(AST), and alkaline phosphatase(ALP) in serum were detected. Pathological injuries of livers were detected by hematoxylin-eosin(HE) staining. The levels of malondialdehyde(MDA), superoxide dismutase(SOD), glutathione peroxidase(GSH-Px), and reduced glutathione(GSH) in the liver were measured. Wes-tern blot method was used to detect the expression of nuclear factor erythroid 2-related factor 2(Nrf2), Kelch-like ECH-associated protein 1(Keap1), peroxisome proliferator-activated receptor gamma, coactivator-1 alpha(PGC-1α), heme oxygenase-1(HO-1), and NAD(P)H quinone dehydrogenase 1(NQO1) in livers. Immunofluorescence was used to detect the expression of Nrf2 and PGC-1α in livers. The results indicated that Astragalus membranaceus/Angelica sinensis ratio of 2∶1 and 5∶1 could significantly reduce the levels of serum AST, ALT, and ALP, improve the pathological damage of liver tissue, increase the levels of GSH and GSH-Px, and reduce the content of MDA in liver tissue. Astragalus membranaceus/Angelica sinensis ratio of 1∶1 and 2∶1 could significantly improve the level of SOD. Astragalus membranaceus and Angelica sinensis before and after compatibility significantly increased the protein expression of HO-1 and NQO1, improved the protein expression of Nrf2 and PGC-1α, and decreased the protein expression of Keap1 in liver tissue. The above results confirmed that the compatibility of Astragalus membranaceus and Angelica sinensis had antioxidant effects by re-gulating Keap1/Nrf2/PGC-1α, and the Astragalus membranaceus/Angelica sinensis ratio of 2∶1 and 5∶1 had stronger antioxidant effect and significantly reduced TP-induced hepatoto-xicity.

Research progress on Huangqi Guizhi Wuwu Decoction and predictive analysis of quality markers / 中国中药杂志

Huangqi Guizhi Wuwu Decoction is a classic prescription in traditional Chinese medicine(TCM) and is known for its effects of tonifying Qi, warming the meridians, and promoting blood circulation to alleviate obstruction. It is primarily used to treat conditions characterized by Qi stagnation, Yang deficiency, and obstruction, and it exhibits pharmacological effects such as immune regulation, anti-inflammation, analgesia, protection of the cardiovascular and cerebrovascular systems, itch relief, reduction of frostbite symptoms, antioxidative stress, promotion of cell apoptosis, and kidney protection. In modern clinical practice, it is commonly used to treat acute myocardial infarction, sequelae of cerebral infarction, cervical spondylosis, frozen shoulder, lower limb arteriosclerosis, lower limb vascular disorders, peripheral neuropathy in diabetes, and lupus nephritis. Recent research has focused on the chemical components, pharmacological effects, and clinical applications of Huangqi Guizhi Wuwu Decoction. Based on the "five principles" of quality markers(Q-markers) in TCM, this study predicted and analyzed the Q-markers of Huangqi Guizhi Wuwu Decoction. It suggested that astragaloside Ⅳ, formononetin, kaempferol, quercetin, cinnamic acid, cinnamaldehyde, 6-gingerol, paeoniflorin, albiflorin, and gallic acid could serve as Q-markers for Huangqi Guizhi Wuwu Decoction. The findings of this study can provide references for quality control of Huangqi Guizhi Wuwu Decoction and the development of new Chinese medicinal formulations.

Effects of Astragalus membranaceus, Peumus boldus and Curcuma longa extracts in diabetic rats / Efeitos de extratos de Astragalus membranaceus, Peumus boldus e Curcuma longa em ratos com diabetes induzida

Objectives: the aim of this study was to evaluate the effects of the association of dry extracts of Astragalus membranaceus, Peumus boldus and Curcuma longa in rats with induced diabetes. Methods: After the induction of type 2 diabetes by intraperitoneal streptozotocin, male Wistar rats were randomly assigned to groups (n=6) and treated for 20 days. The extracts were suspended in water and administered through orogastric gavage once daily as described: Group I: healthy control (saline); group II: received Astragalus membranaceus, Peumus boldus and Curcuma longa (400 mg/kg/day of each dry extract); group III: received Astragalus membranaceus, Peumus boldus, Curcuma longa (400 mg/kg/day of each dry extract) and glibenclamide (15 mg/kg/day). Fasting glucose, glucose tolerance, alanine aminotransferase, aspartate aminotransferase and fructosamine were evaluated. Results: Fasting blood glucose and glucose tolerance for groups II and III were influenced by treatments (p<0.05). The extracts did not significantly influence the efficacy of glibenclamide. Conclusion: The results found in this study allow us to consider that it is not possible to conclude that the compounds evaluated are not effective in DM in rats, due to variables such as total treatment period, doses, size of pancreatic injury caused by streptozotocin, and diet profile may have influenced the results. The studied compounds have potential for application in diabetes and further studies should be carried out to adjust the treatment.

Objetivos: avaliar os efeitos da associação de extratos secos de Astragalus membranaceus, Peumus boldus e Curcuma longa em ratos com diabetes induzida. Métodos: Após a indução de diabetes tipo 2 (DM) por estreptozotocina intraperitoneal, ratos Wistar machos foram distribuídos aleatoriamente em grupos (n=6) e tratados por 20 dias. Os extratos foram suspensos em água e administrados por gavagem orogástrica uma vez ao dia conforme descrito: Grupo I: controle saudável (solução salina); grupo II: recebeu Astragalus membranaceus, Peumus boldus e Curcuma longa (400 mg/kg/dia de cada extrato seco); grupo III: receberam Astragalus membranaceus, Peumus boldus, Curcuma longa (400 mg/kg/dia de cada extrato seco) e glibenclamida (15 mg/kg/dia). A glicemia de jejum, tolerância à glicose, alanina aminotransferase, aspartato aminotransferase e frutosamina foram avaliados. Resultados: A glicemia de jejum e a tolerância à glicose para os grupos II e III foram influenciadas pelos tratamentos (p<0,05). Os extratos não influenciaram significativamente na eficácia da glibenclamida. Conclusão: Os resultados encontrados neste estudo permitem considerar que não é possível concluir que os compostos avaliados não são eficazes no DM em ratos, devido às variáveis como tempo total de tratamento, doses e tamanho da lesão pancreática causada por estreptozotocina, além do perfil da dieta, que podem ter influenciado os resultados. Os compostos estudados têm potencial para aplicação em diabetes e mais estudos devem ser realizados para adequar o tratamento.

Estratégias anti-aging baseadas em moléculas químicas naturais como potentes ativadores de telomerase / Anti-aging strategies based on natural chemical molecules as potent telomerase activators / Estrategias antienvejecimiento basadas en moléculas químicas naturales como potentes activadores de la telomerasa

Mesmo em tempos modernos, os grandes avanços tecnológicos não permitem de forma comprovada retardar o envelhecimento nos seres humanos. Neste sentido, uma das estratégias é o uso moléculas químicas naturais que possuem a ação de ativadores de telomerase, uma vez de que a telomerase é uma ribonucleoproteína transcriptase reversa que possui a função de alongar os telômeros e neutralizar a erosão normal dos telômeros. Neste contexto, este estudo de revisão dedicou-se a aprofundar o conhecimento sobre o uso de moléculas químicas naturais derivadas de plantas que possuem função de ativadores de telomerase para atividade anti-aging. Inúmeras moléculas têm sido propostas e, estudas os seus mecanismos com o intuito de desenvolver novas ferramentas para prevenir/retardar e tratar doenças relacionadas a idade e o envelhecimento. Adicionalmente, o uso de moléculas como ativadores da telomerase têm sido um meio de prolongar o encurtamento dos temoleros, como no caso, de moléculas isolada da erva Astragalus membranaceus (TA-65), curcumina, silbinina e alicina; ademais, outras moléculas de origem natural possuem atividade anti-aging comprovadas, conforme reportadas nesta revisão. Sendo assim, a procura por biomarcadores à base de compostos químicos naturais que estimulem a telomerase, a fim de prolongar a vida dos telômero e assim, retardar o processo de envelhecimento do organismo têm despertado o interesse de diversos pesquisadores ao redor do mundo.

Even in modern times, the great technological advances do not allow in a proven way to delay aging in humans. In this sense, one of the strategies is the use of natural chemical molecules that have telomerase activators, since telomerase is a ribonucleoprotein reverse transcriptase that has the function of lengthening telomeres and neutralizing the normal erosion of telomeres. In this context, this review study was dedicated to deepening the knowledge about the use of natural chemical molecules derived from plants that have telomerase activator function for anti-aging activity. Numerous molecules have been proposed and their mechanisms studied in order to develop new tools to prevent/delay and treat aging-related diseases. Additionally, the use of molecules as telomerase activators has been a means of prolonging the shortening of temolers, as in the case of molecules isolated from the herb Astragalus membranaceus (TA-65), curcumin, silbinin and allicin; in addition, other molecules of natural origin have proven anti-aging activity, as reported in this review. Therefore, the search for biomarkers based on natural chemical compounds that stimulate telomerase in order to prolong the life of telomeres and, thus delay the aging process of the organism has aroused the interest of several researchers around the world.

Aún en los tiempos modernos, los grandes avances tecnológicos no permiten de manera comprobada retrasar el envejecimiento en los humanos. En este sentido, una de las estrategias es el uso de moléculas químicas naturales que tengan activadores de la telomerasa, ya que la telomerasa es una ribonucleoproteína transcriptasa inversa que tiene la función de alargar los telómeros y neutralizar la erosión normal de los telómeros. En este contexto, este estudio de revisión se dedicó a profundizar en el conocimiento sobre el uso de moléculas químicas naturales derivadas de plantas que tienen función activadora de la telomerasa para la actividad antienvejecimiento. Se han propuesto numerosas moléculas y se han estudiado sus mecanismos para desarrollar nuevas herramientas para prevenir/retrasar y tratar enfermedades relacionadas con el envejecimiento. Adicionalmente, el uso de moléculas como activadores de la telomerasa ha sido un medio para prolongar el acortamiento de temolers, como es el caso de moléculas aisladas de la hierba Astragalus membranaceus (TA-65), curcumina, silbinina y alicina; además, otras moléculas de origen natural han demostrado actividad antienvejecimiento, como se reporta en esta revisión. Por ello, la búsqueda de biomarcadores basados en compuestos químicos naturales que estimulen la telomerasa para prolongar la vida de los telómeros y así retrasar el proceso de envejecimiento del organismo ha despertado el interés de varios investigadores a nivel mundial.

Effects of water regulation on biosynthesis of calycosin-7-O-β-D-glucoside in Astragalus membranaceus var. mongholicus / 中国中药杂志

The effects of water regulation on the biosynthesis of calycosin-7-O-β-D-glucoside in 2-year-old Astragalus membranaceus var. mongholicus were studied,and the mechanism was explained from the aspects of key enzyme gene expression and antioxidant enzyme system. The content of calycosin-7-O-β-D-glucoside was determined by HPLC,and the expression levels of six key enzyme genes( PAL,4 CL,CHS,CHI,IFS,13'H) in the synthesis pathway were analyzed by q RT-PCR. The activities of protective enzymes and contents of osmoregulation substances and malondialdehyde were also determined. In the water deficit group,the maximum concentration of calycosin-7-O-β-D-glucoside was 0. 49 mg·g-1 on the 24 th day of treatment. In the whole water regulation,the water deficit group outweighed the water adequate group in osmoregulation substance and MDA contents. The activities of A. membranaceus var.mongholicus antioxidant enzymes SOD,POD,and CAT increased during the initial period of water regulation,but decreased with time.The expression of PAL,CHS,and 13'H in the water deficit group was at a low level,and the 4 CL had active expression,slightly lower than that in the water adequate group. The expression of CHI and IFS elevated rapidly when water deficit occurred. Correlation analysis showed that the content of calycosin-7-O-β-D-glucoside was positively correlated with CHI expression( P<0. 01) and IFS expression( P<0. 05). Therefore,water regulation can change the accumulation pattern of calycosin-7-O-β-D-glucoside,and water deficit may be an effective way to increase its content. CHI and IFS are the key genes in response to water deficit.

Research advance of pharmacological effects of astragalosides on nervous system diseases / 中国中药杂志

Astragali Radix is a traditional Chinese herbal medicine with a long history, which has the functions of tonifying Qi and promoting urination and granulation. Astragalosides are the main effective components of Astragali Radix, and more than 40 triterpenoid saponins have been obtained from Astragalus membranaceus and its related plants, mainly including astragalosides Ⅰ-Ⅷ, isoastragalosides Ⅰ, Ⅱ, and Ⅳ, acetylastragalosides, and soyasaponins. Astragalosides have a wide range of biological activities, such as immunomodulation, antioxidation, and neuroprotection. Nervous system diseases seriously affect people's quality of life, threaten human physical and mental health, and impose a burden on families and society. As natural drugs, astragalosides have good preventive and therapeutic effects on central nervous system diseases. This paper reviews the main pharmacological effects and mechanisms of astragalosides in the treatment of multiple sclerosis, Parkinson's disease, Alzheimer's disease, and cerebral ischemic stroke and proposes the research prospects and potential problems, aiming to provide reference for the clinical application and basic research of astragalosides.

Optimization of analysis methods for Astragali Radix and quality evaluation of standard decoction of Astragali Radix / 中国中药杂志

Astragali Radix is commonly used as bulk medicinal materials. Chinese Pharmacopoeia contains about 150 compound preparations of Astragali Radix, but the sample preparation method under the determination of Astragali Radix content in Chinese Pharmacopoeia is tedious and time-consuming, not convenient for the test of a large number of samples. Therefore, it is of great significance to simplify the sample preparation method and improve the practicability of the method for the quality control of Astragali Radix and its preparations. In this study, ultrasonic extraction method was used instead of heated reflux extraction, and solid phase extraction method was used to enrich and prepare the samples. A set of practical quality evaluation method was established for Astragali Radix slices and standard decoction, greatly shortening the sample preparation time and improving the accuracy of the method. The results of Astragali Radix standard decoction analysis showed that the transfer rate of calycosin 7-O-β-D-glucospyranoside,(96.5±28.7)%, had great variation, which was found to be related to the conversion of mulberry isoflavone glucoside into calycosin 7-O-β-D-glucospyranoside during the preparation of standard decoction. The transfer rates were(59.4±14.4)% and(101.3±12.3)% for calycosin and astragaloside Ⅳ respectively, which were relatively stable. Therefore, it is suggested that Astragali Radix slices and water decoction preparations should be evaluated by using calycosin and astragaloside Ⅳ as the quality evaluation index. The results provide a scientific and practical method for quality control of Astragali Radix slices and its standard decoction, and also provide scientific evidence for quality evaluation of the preparations.

Comparison of 12 constituents of Astragali Radix directionally processed with organic acid by UPLC-MS / 中国中药杂志

To establish an UPLC-MS method for the simultaneous content determination of 4 saponins and 8 flavonoids, in order to analyze the effect of Astragali Radix directionally processed with organic acid on the content of glycosides and aglycones. The separation was carried out on ACQUITY UPLC HSS T_3(2.1 mm×100 mm, 1.8 μm), the mobile phase was eluted with the mixture of 0.1 mol·L~(-1) formic acid water solution and 0.1 mol·L~(-1) formic acid acetonitril in a gradient mode. The detection wavelength was 260 nm, the flow rate was 0.5 mL·min~(-1), the column temperature was 30 ℃, and the injection volume was 2 μL. Mass spectrometry analysis was performed with an electrospray ionization(ESI) source in a positive ion mode. The 12 constituents showed good linear relations within their own ranges(R~2≥0.999 2),with good average recoveries. The results showed no significant change in saponins but both qualitative and quantitative changes in flavonoids after directional processing of Astragali Radix with organic acid. The established method can provide methodological reference for analyzing the effect of Astragali Radix directionally processed with organic acid on glycosides and aglycones.

Study on protective effect of vanillic acid from Astragalus membranaceus on hypertensive cardiac remodeling based on network pharmacology screen / 中国中药杂志

To identify and verify the active ingredients from Astragalus membranaceus on hypertensive cardiac remodeling based on network pharmacology and heart RNA-sequencing data. The monomers of A. membranaceus and their intervention target database were established by using network pharmacology. The genes associated to cardiac remodeling were then screened by analyzing cardiac RNA-sequencing data. An overlap between genes related to cardiac remodeling and targets of ingredients form A. membranaceus was collected to obtain monomers with protective effect on hypertensive cardiac remodeling. Angiotensin Ⅱ(AngⅡ)-induced mouse cardiac remodeling model was used to validate the protective effect of active ingredients from A. membranaceus on hypertensive cardiac remodeling. Finally, a total of 81 monomers and 1 197 targets were enrolled in our database. Mouse RNA-sequencing data showed that 983 genes were significantly up-regulated and 465 genes were down-regulation in myocardial tissues of the cardiac remodeling mice as compared with blank group mice, respectively. Ninety-two genes were found via overlapping between genes related to cardiac remodeling and targets, involving 59 monomers from A. membranaceus. Further research found that vanillic acid(VA) could intervene 27 genes associated with hypertensive cardiac remodeling, ranking top 1. Meanwhile, VA could significantly inhibit AngⅡ-induced increase in ratio of heart weight to body weight and heart weight to tibial length, ANP and BNP mRNA levels in myocardial tissues, myocardial tissue damage, cardiac fibrosis level and cardiac hypertrophy level in vivo. Those results showed that network pharmacology screen-based VA has protective effect on AngⅡ-induced cardiac remodeling.

Grey correlation analysis and path analysis between isoflavones content in Astragali Radix and climate factors / 中国中药杂志

In this study, HPLC was used to determine the content of the four isoflavones of Astragalus membranceus var. mongholicus from different regions(calycosin-7-glucoside, ononin, calycosin and formononetin), and gray correlation analysis and path analysis were used to explore the influence of climate factors on the content of isoflavone components in A. membranceus var. mongholicus. The results showed that there were significant differences in the content of the four isoflavones in different areas(P<0.05); grey correlation analysis showed that the highest temperature in July, the lowest temperature in January and the daily average temperature had a greater impact on the content of flavonoid glycosides, meanwhile precipitation and relative humidity were the more important factors for the accumulation of flavonoid aglycones. According to the general analysis, the direct positive effects of the lowest temperature in January and altitude on the contents of four isoflavones in A. membranceus var. mongholicus were significant. High altitude and extreme temperature conditions might be more adverse to the formation and accumulation of isoflavone components. Therefore, the religions of A. membranceus var. mongholicus with high contents of isoflavones should be chosen the low altitude region with higher minimum temperature in January. This study provides a reference basis for the quality evaluation of A. membranceus var. mongholicus, and basic data for the selection of suitable habitat, construction of planting standards and directional cultivation of medicinal materials in A. membranceus var. mongholicus.

Analysis of quality difference based on Astragalus membranaceus var. mongholicus in genuine region / 中国中药杂志

This work is to establish the fingerprint of Astragalus membranaceus var. mongholicus by HPLC-ELSD method, and to analyze the simulated wildness degree of A. membranaceus var. mongholicus in the genuine region of Inner Mongolia, Ningxia and Gansu. Compared with wild A. membranaceus var. mongholicus, the quality differences of A. membranaceus var. mongholicus in the genuine region were analyzed by identification of chromatographic peaks and similarity evaluation, cluster analysis(CA), principal components analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA). HPLC fingerprints of A. membranaceus var. mongholicus in different genuine regions are established. The qualitative analysis of mass spectrometry identified 18 components. The similarity evaluation shows that the similarity of 32 batches of A. membranaceus var. mongholicus samples was 0.688-0.993. Among them, the similarity of samples in Shanxi, Inner Mongolia, Ningxia is 0.688-0.993, 0.835-0.989, 0.934-0.988, respectively and the similarity of samples in Gansu is 0.729-0.876 except No. 25 sample. The results of CA show that the samples of A. membranaceus var. mongholicus can be grouped into four categories according to the production area except the No. 11 and No. 25 samples. The results of PCA indicate that 32 batches of A. membranaceus var. mongholicus samples can be clustered according to quality and origin, and the quality of A. membranaceus var. mongholicus in Inner Mongolia is the closest to the wild breed. The results of OPLS-DA indicate that there are six components that can distinguish the wild and domestic A. membranaceus var. mongholicus, which are malonylastragaloside Ⅰ, astragaloside Ⅰ, calycosin-7-O-β-D-glycoside-6″-O-malonate, calycosin-7-O-β-D-glycoside, formononetin-7-O-β-D-glycoside-6″-O-malonate, and astrapterocarpan-3-O-β-D-glycoside-6″-O-malonate. The established method can be used to analyze differences between A. membranaceus var. mongholicus origin and planting environment, and can provide references for the protection and replacement of wild A. membranaceus var. mongholicus resources, and the cultivation, processing and production of A. membranaceus var. mongholicus.

Establishment of health risk assessment model for assessing medicinal and edible plants contaminated by heavy metals--take Astragali Radix,Codonopsis Radix and Laminariae Thallus as examples / 中国中药杂志

With the improvement of living standard,the theory of " medicine and food homology" has developed rapidly in the field of diet,medicine and health preservation. In recent years,many literatures have been reported on the active ingredients and pharmacological effects of medicinal and edible plants,but relatively few reports have been reported on their safety investigation. Therefore,to further evaluate the quality and safety of medicinal and edible plants,Astragali Radix,Codonopsis Radix and Laminariae Thallus were selected as our research objects in this study. Moreover,the pollution level and the potential health risk of heavy metals were deeply assessed in different types of medicinal and edible plants. Especially,the contents of chromium,copper,arsenic,cadmium,mercury and lead in these three herbs were determined by inductively coupled plasma mass spectrometry( ICP-MS),and their health risk level was evaluated by target hazard coefficient method. The results showed that under the international heavy metal limit standard( ISO 18664-2015,GB 2762-2017),the over-standard rates were 25%,77% and 100% in 16 batches of Astragali Radix,26 batches of Codonopsis Radix and 9 batches of Laminariae Thallus,respectively. Besides,the values of target hazard quotients( THQ) for adults and children are 0. 028 244,0. 063 505 and 0. 014 485,0. 032 568 in Astragalus membranaceus and Codonopsis pilosula,respectively,which were higher than the standard values of 0. 02 and 0. 011 25. While,the total heavy metals THQ values for adults and children are 0. 023 734 and 0. 020 287 in Laminariae Thallus,which were much higher than the standard values of 0. 008 0 and 0. 007 5. However,the CR values of As,Cd and Pb in the three herbs were lower than 1×10~(-6). Above results indicated that those six harmful elements have certain health hazards to the exposed population,but there is no potential carcinogenic effect. It can be seen that,there were still presence of the pollution of harmful elements,and it is necessary to establish the reasonable limit standards and quality control methods of medicinal and edible plants in time.

Responses of seed germination of Astragalus membranaceus to light and temperature conditions accompanied with drought and salt stresses / 中国中药杂志

We studied the seed germination of Astragalus membranaceus under PEG and Na Cl osmotic stress gradients( 0,-0. 1,-0. 3,-0. 5,-0. 7 MPa) respectively applied with light( continuous light,light 12 h/dark 12 h circulation and continuous dark) and temperature( constant 15 ℃,15 ℃ 12 h/30 ℃ 12 h circulation and constant 30 ℃) treatments. The results showed as following: ① Under the light and temperature interactive treatments,total germination percentage( TGP) was restrained by high temperature and continuous light also decreased TGP under high temperature. Mean germination time( MGT) was not changed by light mode. Root development was enhanced by dark and low temperature. Shoot development was enhanced by light and high temperature. Hypocotyl length was enhanced by dark and high temperature. ② Under the light and temperature interactive treatments combined respectively with PEG and NaCl stress conditions,although the inhibitions of seed germination and growth were gradually strengthened with the increases of osmotic stresses,slight osmotic stress can promote seed germination. Under the same osmotic potential,the effects of PEG on TGPs and MGTs were stronger than that of NaCl. As the temperature increase,the seeds may change from photo-neutrality to photo-phobia. Decreased TGP under drought and continuous light interactive treatment is an adaptation strategy to avoiding drought. Hypocotyl growth accelerated under continuous dark treatment is an ecological trait which could increase dry matter input in stem and height for more light. Seed development under high concentration of NaCl treatment is better than that of PEG treatment due to low water potential caused by Na~+,which can enter into seed coat and promote water absorption.

Safety evaluation of heavy metals contaminated Astragalus membranaceus using health risk assessment model / 中国中药杂志

To evaluate the safety of heavy metals contaminated Astragalus membranaceus,an appropriate protocol was established to study the heavy metals pollution level by health risk assessment. This study provided a detailed procedure to assess the medicinal herbs in quality control and safety evaluation,and expected to create awareness among the public on the safety of consuming of A. membranaceus or any other kinds of medicinal herbs. The heavy metals content of Cu,As,Cd,Pb and Hg in a total of 45 batches of A. membranaceus were carefully analyzed with a developed inductively coupled plasma mass spectrometry( ICP-MS). Besides,the heavy metal contamination level was further evaluated through 4 main assessment parameters,including maximum residue limit( MRL) set by International Standard Organization,estimated daily intake( EDI) set by IUPAC,target hazard quotients( THQ) and Total THQ set by USEPA and total THQs in raw herbs of A. membranaceus. In addition,the recommended MRLs of 5 main heavy metals aimed to A. membranaceus were calculated based on the regulated consumption quantity. The result showed that,under the ISO international standard of Chinese medicine-Chinese herbal medicine heavy metals,the unqualified rate was 8. 89% for A. membranaceus,which including 4 batches of A. membranaceus exceeded the MRL of As. Here,the standard THQ value of A. membranaceus was firstly proposed as 0. 02 and 0. 011 25 for adults and children,respectively,which were calculated with the recommended consumption quantity of 30 g and 9 g for adults and children. Furthermore,the values of THQ for As and total THQs in adults and children were exceeded the standard THQ in A. membranaceus,and the recommended MRLs of Pb,Cd,Hg and Cu in above medicinal materials that calculated based on health risk assessment model were higher than the regulated MRLs that set by ISO and Chinese Pharmacopeia. The research showed that the contents of heavy metals in A. membranaceus were not in the safe range and the certain non-carcinogenic risks to human body cannot be neglected. Based on above investigation result,it is easily known that the common evaluation method for raw herbs based on the comparison of MRL of heavy metals was not precise enough,and the international model of health risk assessment should be built for each medicinal herb. Above all,this study provided a more realistic research approach for safety evaluation of any other kinds of heavy metals contaminated medicinal herbs,including the establishment of heavy metals standard limit in a specified medicinal herb under recommended consumption quantity,and it is expected to create awareness among the public on the safety of consuming any other medicinal herbs.

Astragalus membranaceus improves therapeutic efficacy of asthmatic children by regulating the balance of Treg/Th17 cells / 中国天然药物

Astragalus membranaceus may be a potential therapy for childhood asthma but its driving mechanism remains elusive. The main components of A. membranaceus were identified by HPLC. The children with asthma remission were divided into two combination group (control group, the combination of budesonide and terbutaline) and A. membranaceus group (treatment group, the combination of budesonide, terbutaline and A. membranaceus). The therapeutic results were compared between two groups after 3-month therapy. Porcine peripheral blood mononuclear cells (PBMCs) were isolated from venous blood by using density gradient centrifugation on percoll. The levels of FoxP3, EGF-β, IL-17 and IL-23 from PBMCs and serum IgE were measured. The relative percentage of Treg/Th17 cells was determined using flow cytometry. The main components of A. membranaceus were calycosin-7-O-glucoside, isoquercitrin, ononin, calycosin, quercetin, genistein, kaempferol, isorhamnetin and formononetin, all of which may contribute to asthma therapy. Lung function was significantly improved in the treatment group when compared with a control group (P < 0.05). The efficacy in preventing the occurrence of childhood asthma was higher in the treatment group than the control group (P < 0.05). The levels of IgE, IL-17 and IL-23 were reduced significantly in the treatment group when compared with the control group, while the levels of FoxP3 and TGF-β were increased in the treatment group when compared with the control group (P < 0.05). A. membranaceus increased the percentage of Treg cells and reduced the percentage of Th17 cells. A. membranaceus is potential natural product for improving the therapeutic efficacy of combination therapy of budesonide and terbutaline for the children with asthma remission by modulating the balance of Treg/Th17 cells.

Changes in the constituents and UV-photoprotective activity of Astragalus membranaceus caused by roasting / 한국영양학회지

PURPOSE: Astragalus membranaceus (AM) is an important traditional medicinal herb. Pharmacological research has indicated that AM has various physiological activities such as antioxidant, anti-inflammatory, immunoregulatory, anticancer, hypolipidemic, antihyperglycemic, and hepatoprotective activities. The bioactive substances responsible for the physiological activities in AM, including many antioxidant substances, change during the roasting process. This study investigated and compared the changes in the antioxidant constituents of AM caused by roasting. METHODS: DPPH (1,1-diphenyl-2-picryl hydrazyl) and ABTS⁺ (2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt) radical scavenging activities and their total phenolic content (TPC) were measured. High-performance liquid chromatography (HPLC) analysis was performed to confirm any changes in the isoflavonoids of roasted AM (R-AM),. The cell viability of UVB-induced HDF (Human dermal fibroblast) cells treated with AM and R-AM extracts was investigated. The comet assay was used to examine the inhibitory effects of R-AM extracts on DNA damage caused by oxidative stress. RESULTS: The DPPH and ABTS⁺ radical scavenging activities were 564.6±20.9 and 108.2±3.1 (IC₅₀ value) respectively, from the 2R-AM. The total phenol content was 47.80±1.40 mg GAE/g from the 1R-AM. The values of calycosin and formononetin, which are the known isoflavonoid constituents of AM, were 778.58±2.72 and 726.80±3.45 µg/g respectively, from the 2R-AM. Treatment of the HDF cells with R-AM (50 ~ 200 µg/mL) did not affect the cell viability. Furthermore, the R-AM extracts effectively protected against UVB-induced DNA damage. CONCLUSION: The findings of this study indicate that R-AM increases its isoflavonoid constituents and protects against UVB-induced DNA damage in HDF cells.

Metabolomics analysis and rapid identification of changes in chemical ingredients in crude and processed Astragali Radix by UPLC-QTOF-MS combined with novel informatics UNIFI platform / 中国天然药物

Astragali Radix, the root of Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao or Astragalus membranaceus (Fisch.) Bge., is widely used as a tonic decoction pieces in the clinic of traditional Chinese medicine (TCM). Astragali Radix has various processed products with varying pharmacological actions. There is no modern scientific evidence to explain the differences in pharmacological activities and related mechanisms. In the present study, we explore the changes in chemical components in Astragali Radix after processing, by ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) combined with novel informatics UNIFI platform and multivariate statistical analysis. Our results showed that the crude and various processed products could be clearly separated in PCA scores plot and 15 significant markers could be used to distinguish crude and various processed products by OPLS-DA in UNIFI platform. In conclusion, the present study provided a basis of chemical components for revealing connotation of different processing techniques on Astragali Radix.