BACKGROUND

Radix Astragali, one of most widely used and important traditional Chinese medicines, is cultivated in different geographical regions. Because of varying growing conditions, the qualities of Radix Astragali vary, which can give rise to differences in clinical therapy. Detecting adulteration is a routine requirement in pharmaceutical practice.

OBJECTIVE

To develop a simple and accurate approach to discriminate the geographical origin and potential adulteration of Radix Astragali, derived from the root of Astragalus membranaceus (Fischer) Bunge var. mongholicus (Bunge) Hsiao, using Fourier transform infrared (FT-IR) spectroscopy and chemometric methods.

METHODS

To obtain characteristic IR spectra for accurate discrimination, a one-solvent extraction method was utilised following a novel evaluation method for selecting appropriate solvents. Samples of Radix Astragali from different geographical origins were discriminated using FT-IR spectroscopy and discriminant partial least squares (DPLS) methods. FT-IR spectroscopy combined with Mahalanobis distance was employed to detect adulteration of Radix Astragali.

RESULTS

In comparison with other solvents, butanone was more effective at extracting samples. Radix Astragali samples were accurately assigned to their corresponding geographical origins by using FT-IR spectroscopy and DPLS method. Most adulterated samples were detected accurately by application of FT-IR spectroscopy combined with Mahalanobis distance.

CONCLUSIONS

FT-IR spectroscopy combined with chemometric method was developed and demonstrated to be a useful tool to discriminate geographical origin and adulteration of Radix Astragali.

Astragali Radix (AR) is one of the most popular herbal medicines in traditional Chinese medicine (TCM). Wild AR is believed to be of high quality, and substitution with cultivated AR is frequently encountered in the market. In the present study, two types of ARs (wild and cultivated) from Astragalus membranaceus (Fisch.) Bge. and A. membranaceus var. mongholicus (Bge.) Hsiao, growing in different regions of China, were analyzed by NMR profiling coupled with multivariate analysis. Results showed that both could be differentiated successfully and cultivation patterns or growing years might have greater impact on the metabolite compositions than the variety; the metabolites responsible for the separation were identified. In addition, three extraction methods were compared and the method (M1) was used for further analysis. In M1, the extraction solvent composed of water, methanol, and chloroform in the ratio of 1 : 1 : 2 was used to obtain the aqueous methanol (upper layer) and chloroform (lower layer) fractions, respectively, showing the best separation. The differential metabolites among different methods were also revealed. Moreover, the sucrose/glucose ratio could be used as a simple index to differentiate wild and cultivated AR. Meanwhile, the changes of correlation pattern among the differential metabolites of the two varieties were found. The work demonstrated that NMR-based non-targeted profiling approach, combined with multivariate statistical analysis, can be used as a powerful tool for differentiating AR of different cultivation types or growing years.

OBJECTIVE

To study the effect of astragalus membranceus(fisch) bge, codonpsis pilosula and glycyrrhiza uralensis fisch on airway responsiveness.

METHODS

FVC, FEV1 and PEF of twenty-eight asthmatic patients were measured before and six weeks after treated with Astragalus membranceus(fisch) bge, codonpsis pilosual and glycyrrhiza uralensis fisch; methacholine challenge test with 0.033 mumol and 1.98 mumol.

RESULTS

After 28 asthmatic patients received the treatment of those herbs, the results showed that FVC value increased obviously (t = 2.217, P < 0.05) and the value of FEV1 and PEF was all higher compared with before treatment of the herbs (P < 0.01).

CONCLUSIONS

The herbal medicine including astragalus memberanaceus (fisch) bge, codonpsis pilosual and glycyrrhiza uralensis fisch can decrease the airway responsiveness of the patients with asthma.

OBJECTIVE

To make a comparative study on wild and cultivated Astragali Radix in Wuchuan, Neimenggu where is one of the geo-authentic producing areas of Astragalus membranaceus var. mongholicus.

METHODS

This comparative study focus on shapes and properties, microscopic features of transverse section and powder of roots, qualitative evaluation of wild and cultivated Astragali Radix.

RESULTS

Wild Astragali Radix had a cylindrical main root, 2 or 3 root branches, dark brown color and many lenticels on the root bark. Cultivated Astragali Radix had a long cylindrical root, few root branches, yellowish white or light brown and fewer lenticels on the root bark. The differences of microscopic features were that the number of cork cells layers in wild Astragali Radix was bigger than that in cultivated Astragali Radix; stone cells were only observed in wild Astragali Radix; distinct annual rings in the xylem were only existed in cultivated Astragali Radix. The results of qualitative evaluation reveal that the contents of major active isoflavonoids and saponins in wild Astragali Radix are higher than those in cultivated Astragali Radix.

CONCLUSIONS

There are some diagnostic differences in the main microscopic features of transverse section and powder between wild and cultivated Astragali Radix. The contents of major active isoflavonoids and saponins in wild Astragali Radix are higher than those in cultivated Astragali Radix. Our study provides important scientific evidence for reasonable and effective uses of wild and cultivated Astragali Radix in Wuchuan, and also provides a reliable basis for the quality control of Astragali Radix.

Astragalus membranaceus (AM) provides a rich source of polysaccharides that can act as powerful antioxidants, but their potential as feed ingredients in the lamb industry still rarely exploited. The objective of this study was to investigate the effect of dietary astragalus membranaceus supplementation on oxidative stability of goat muscles. Longissimus dorsi (LD) muscles from two groups of Cashmere goat (basal diet, C group; basal diet supplemented with 1% astragalus membranaceus root, AM group) were evaluated for lipid oxidation, myoglobin oxidation, activity of antioxidant enzymes, and antioxidant capacity. The results showed that color parameters in Cashmere goat of two feeding conditions were no significant difference (p > .05). In AM group, myoglobin (Mb) content was higher than C, while metmyoglobin (MMb) (p < .05) and malondialdehyde (MDA) (p < .01) were lower. Additionally astragalus membranaceus supplementation had a significant effect on superoxide dismutase (SOD) and catalase (CAT) (p < .001). In whole, the AM group goats presented a relatively higher antioxidant capacity than C. Especially, RSA and CUPRAC values of AM group goats had significantly higher than C (p < .05). Consequently, the AM group goats ingested abundant astragalus membranaceus, which enhanced the antioxidant capacity. Thus, it can eliminate free radicals and effectively inhibit oxidation.

Keywords: Cashmere goat; antioxidant ability; antioxidant enzymes; grazing style; oxidation.

Astragalus polysaccharide (APS), a common Chinese herbal compound extracted from Astragalus membranaceus, has been proposed to increase the tumour response of and stabilize chemotherapy drugs while reducing their toxicity. Here, we examined the effects of APS on apoptosis in gastric cancer (GC) cells in the presence or absence of adriamycin (0.1 µg/mL).

GC cells cultured in the presence or absence of adriamycin (0.1 µg/mL) were administered APS (50-200 µg/mL) for 24-72 h and subjected to an MTT assay to examine cell viability. Active caspase-3 expression and DNA fragmentation were assessed to evaluate apoptosis, and real-time PCR was used to analyse the expression levels of multidrug resistance (MDR1) genes and tumour suppressor genes. Western blot analysis was applied to detect cleaved caspase-3 and phosphorylated AMPK (p-AMPK).

Results

Cellular viability was profoundly reduced by APS, and GC cell apoptosis was strongly increased by APS in a time- and dose-dependent manner; these changes may be linked to an increase in p-AMPK levels because the AMPK inhibitor compound C blocked the effects of APS. Similarly, adriamycin-induced decreases in cellular viability and apoptosis of GC cells were enhanced by APS administration. The expression of tumour suppressor genes (SEMA3F, P21^WAF1/CIP1, FBXW7), but not of MDR1, was increased by APS compared to the control, and p-AMPK levels were lower in adriamycin-resistant GC cells than in either adriamycin-sensitive GC cells or an immortalized human gastric epithelial cell line.

APS induces apoptosis independently and strengthens the proapoptotic effect of adriamycin on GC cells, suggesting that APS may act as a chemotherapeutic sensitizer.

Alterations in dopamine (DA) and serotonin (5-HT) transmission have been implicated in the pathophysiology of attention deficit/hyperactivity disorder (ADHD). We have previously reported that juvenile rats with neonatal habenula lesion (NHL) exhibit an assortment of behavioral alterations resembling ADHD symptoms. In this study, we investigated the impacts of NHL on DA and 5-HT transmission in mesocorticolimbic regions of rats. Male Sprague-Dawley rats with microinjection of ibotenic acid into the habenula at postnatal day (PND) 7 were subjected for a battery of locomotion test, object exploration test and delay discounting test in the juvenile period (PND28-35), followed by DA and 5-HT brain tissue concentration measurements using high-performance liquid chromatography (HPLC). NHL rats exhibited hyperlocomotion, impulsivity, and attention deficits. NHL induced alterations of tissue DA and 5-HT concentrations only in some mesocorticolimbic regions. However, positive correlations, indicating the balance, between DA and 5-HT observed in control (CTR) rats, were more extensively disrupted across mesocorticolimbic regions in NHL rats. Pharmacological manipulations that modulated both DA and 5-HT systems simultaneously with Astragalus membranaceus (AM) and its active compound formononetin (FOR) normalized the NHL-induced DA and 5-HT imbalance in several brain areas, which consequently improved the behavioral alterations. These results suggest that behavioral alterations caused by NHL may be associated with mesocorticolimbic DA/5-HT imbalance. Drug treatments targeting multiple monoamine systems may be useful to improve the NHL-induced changes.

Keywords: ADHD; Animal model; Habenula; Monoamine; Natural medicine; Neurodevelopment.

We aimed to identify the optimum mixing ratio for various ingredients to obtain complex extracts with high extract yields and immune-enhancing activity in animals. The extract yield and amounts of nitric oxide (NO) and interleukin (IL)-6 were set to maximum for modeling predictions. The predicted optimum values for the mixing ratio were 49.5% for Rhodiola crenulata, 26.1% for Astragalus membranaceus, and 24.4% for Panax quinquefolius, and the predicted response values were 31.5% yield, 13.4% NO production, and 6.1% IL-6 production; actual values were 35.3% yield, 14.7% NO, and 6.6% IL-6. The optimum mixing ratio extract (OMRE) was used for the animal experiments. Treating mice with OMRE at 200 mg/kg produced significant increases in spleen indexes and T-cell/B-cell proliferation. OMRE treatment increased IL-10 and IL-6 production in concanavalin A- and lipopolysaccharide-induced T- and B- lymphocytes, respectively. These results provide a basis for the development of functional extracts and drinks.

Aim and objective: Cell death is a main pathological change in brain ischemia. Astragalus membranaceus (Ast) and ligustrazine (Lig), as traditional Chinese herbs, have a protective effect against ischemia-reperfusion injury.we aim to find whether the underlying protective mechanism of Astragalus membranaceus and ligustrazine against Oxygen-glucose deprivation/reoxygenation (OGD/R) -induced injury in RBMECs is related to PKCδ/MARCKS pathway.

Materials and methods: OGD/R preconditioning was instituted in rat brain microvascular endothelial cells (RBMECs). The survival and apoptosis of RBMECs were detected by a Cell Counting Kit-8 and TUNEL staining; PKCδ/MARCKS and MMP9 expression were examined by immunofluorescence, western blot and quantitative real-time PCR.

Results: OGD/R stimulation significantly increased RBMEC apoptosis, whereas Ast+Lig, Rottlerin or Ast+Lig+Rottlerin treatment evidently reduced cellular apoptosis, and increased cell viability (P <0.05). Furthermore, Ast+Lig, Rottlerin or Ast+Lig+Rottlerin treatment significantly reduced mRNA expression levels of PKCδ/MARCKS and MMP9 (P <0.05), compared to OGD/R control group. Moreover, Ast+Lig, Rottlerin or Ast+Lig+Rottlerin treatment evidently reduced protein expression levels of PKCδ, MMP9, and MARCKS (P <0.05), compared to OGD/R control group, detected by western blot-ting or immunofluorescence.

Conclusions: The administration of Astragalus membranaceus and ligustrazine protected RBMECs against OGD/R-induced apoptosis. PKCδ/MARCKS and MMP9 expression were significantly increased after OGD/R stimulation, while Astragalus membranaceus and ligustrazine treatment evidently suppressed. Collectively,Astragalus membranaceus and ligustrazine play protective effects against OGD/R-induced injury in RBMECs through regulating PKCδ/MARCKS pathway to inhibit MMP9 activation.

Keywords: Astragalus membranaceus; Injury; Ligustrazine; Oxygen–glucose deprivation/reoxygenation; PKC δ/MARCKS; Rat brain microvascular endothelial cell..

Astragalus roots from Astragalus membranaceus Bunge or Astragalus membranaceus var. mongholicus (Bunge) Hsiao are among the most popular traditional medicinal plants due to their diverse therapeutic uses based on their tonic, antinephritic, immunostimulant, hepatoprotectant, diuretic, antidiabetic, analgesic, expectorant and sedative properties. Currently, the herb is produced or cultivated in various sites, including 10 different locations in China with very diverse environmental conditions. These differences affect their metabolic pools and consequently their medicinal properties. The comparative metabolic profiling of plants of different geographical origins or ages could contribute to detect biomarkers for their quality control and thus guarantee the efficacy of the herbal medicines produced with this drug. In this paper nuclear magnetic resonance spectroscopy (NMR)-based metabolomics was applied for to plants of different origins and age for this purpose. The results of this study show that in the set of samples evaluated, age is more discriminating than geographical location. The quantity of individual flavonoids and some primary metabolites contributed most to this age differentiation. On the other hand, based on the analysis of orthogonal partial least square (OPLS) modeling, the marker metabolites for the geographical origin were saponins and isoflavonoids.

Endometritis, inflammation of the endometrium, is a common reproductive obstacle disease that can lead to infertility in female animals. Astragaloside IV (AS IV), one of the major and active components of the Astragalus membranaceus (Fisch.) Bunge, is known for its anti-inflammatory effects. In the present study, the effects and mechanisms of AS IV on lipopolysaccharide (LPS)-induced endometritis were investigated using a mouse model. Female mice were prepared with AS IV (0.01 mg/g) by gavage for six days before being stimulated with LPS. The results showed that the histopathological changes, levels of inflammatory cytokines (IL-1β and TNF-α), concentration of NO, and myeloperoxidase (MPO) activity in LPS-induced uteri were attenuated significantly by pretreatment with AS IV. Furthermore, LPS-induced activations of NF-κB, p38, and JNK signal pathways were suppressed by pretreatment with AS IV. In conclusion, the data provided new evidence that AS IV effectively attenuates LPS-induced endometritis through inhibition of TLR4-mediated NF-κB, p38, and JNK signaling pathways, implying that AS IV might become a promising potential anti-inflammatory agent for endometritis and other inflammatory diseases.

Cigarette smoking-related lung injury is one of the most common and fatal etiologies of many respiratory diseases, for which no effective interventions are available. Astragaloside Ⅳ (ASⅣ) is an active component extracted from Astragalus membranaceus. It is prescribed as a treatment for upper respiratory tract infections. Here, we report the potential anti-inflammatory effects and mechanisms of ASⅣ on cigarette smoking extract- (CSE)-exposed RAW264.7 cells. Murine macrophages were exposed to CSE, followed by administration of ASⅣ at 25-100 μg/mL for 24 h. ASⅣ significantly rescued CSE-induced cell death by inhibition of release pro-inflammatory cytokines. We measured autophagy as an intracellular scavenger by analyzing autophagic flux using tandem mRFP-GFP-LC3 fluorescence microscopy. Following administration with ASⅣ in CSE-exposed RAW264.7 cells, there was a notable increase in autophagosomes and a range of autophagic vacuoles were generated, as seen with transmission electron microscopy. Loss of autophagy following transfection siRNA aggravated inflammatory injury and release of inflammatory cytokines. Mechanistically, ASⅣ-triggered autophagy is mediated by the TLR4/NF-κB signaling pathway to reduce inflammation. Taken together, our findings suggest that ASⅣ acts stimulates autophagy, and that ASⅣ induces autophagy by inhibiting the TLR4/NF-κB signaling pathway, contributing to alleviation of inflammation.

Keywords: Astragaloside Ⅳ; Autophagy; Inflammation; TLR4/NF-κB signaling pathway.

Astragalus membranaceus root, Huang Qi in Chinese, is a popular medicinal herb traditionally used to regulate blood glucose. Herein, the identification and characterization of two families of cysteine-rich peptides (CRPs), designated α- and β-astratides, from A. membranaceus roots are reported. Proteomic analysis showed that α-astratide aM1 and β-astratide bM1 belong to two distinct CRP families. The six-cysteine-containing and proline-rich α-astratide aM1 displayed high sequence identity to Pea Albumin 1 Subunit b (PA1b), while the eight-cysteine-containing β-astratide bM1 showed sequence similarity to plant defensins. An antifungal assay revealed that bM1 possessed potent antifungal activity. In contrast, aM1 showed a cytotoxic effect against insect Sf9 cells. More importantly, aM1 decreased insulin secretion in mouse pancreatic β cells, suggesting it could interfere in glucose homeostasis, which accounts for the adaptogenic property of A. membranaceus. Phylogenetic clustering analysis suggested that the proline-rich aM1 is a putative prolyl oligopeptidase inhibitor and belongs to a novel subfamily of PA1b-like peptides, while bM1 belongs to a new subfamily of plant defensins. Together, the study reveals that astratides are multifunctional CRPs in plants, which expand the existing library of PA1b-like peptides and plant defensins and further our understanding of their roles in host-defense system and leads as peptidyl therapeutics.

Effect of Jian Yan Ling (JYL, mainly consisted of Succinum, Pteria margaritifers and Astragalus membranaceus etc.) on serum lipids was studied by double-blind method in 128 hyperlipemia patients. Patients were divided randomly into two groups, the test groups (64 cases) treated with JYL, while the other 64 cases treated with placebo as control for 3 months. Serum total cholesterol (TC), triglyceride (TG), TC-high density lipolipid-cholesterol (TC-HDL-C), apoproteins were observed before and after treatment. Results showed that the TC, TG, apoproteins and lipoprotein-a of test group were all dropped significantly after JYL treatment (P < 0.01). The results suggested that JYL could regulate the blood lipids and could also exert an effect of antiatherosclerosis through reducing blood lipoprotein-a level. It is worth while to use JYL as an antihyperlipemic agent clinically.

Reactivation of Toxoplasma gondii infections and serious clinical manifestations such as encephalitis may develop in immunocompromised subjects and AIDS patients. Different protocols are used for the treatment of toxoplasmosis in high-risk patient groups, however life-long prophylactic therapy against reactivation risk in AIDS patients may lead to several undesired results. Atovaquone is an effective antiprotozoal agent against toxoplasmosis with minor side effects. On the other hand, Astragalus membranaceus root extract (AmE) has been shown to have immunomodulatory and antimicrobial activities, empowering immunity by enhancing proliferation and activation of phagocytic cells mainly macrophages, and inducing Th1 type immune response. The aim of this study was to investigate the effectiveness of atovaquone alone and in combination with AmE, in the treatment of toxoplasmosis, and on the levels of IL-2, IL-12 and IFN-γ in experimentally infected mice with T.gondii. For this purpose, four experimental groups, each consisting of eight BALB/c mice, were set with the approval of Ethics Committee for the Animal Experiments. All the mice were infected with 0.5 ml of a suspension containing 2 x 104/ml trophozoites prepared from T.gondii RH strain by intraperitoneal injection. Twenty-four hours after the infection, atovaquone (100 mg/kg/day) was given to atovaquone group, AmE (0.075 mg/g) to astragalus group and atovaquone (100 mg/kg/day) plus AmE (0.075 mg/g) to Atovaquone + Astragalus (Ato + Astra) group by oral gavage. The mice in the fourth group, which was the control group, were all infected but untreated. The above administrations were carried out for seven days. On the 8th day peritoneal fluids of mice were collected under anaesthesia and trophozoite numbers per 1 ml were detected by counting on the Thoma slide. In addition, the heart bloods of mice were drawn and IL-2, IL-12, IFN-γ levels were determined in serum samples by using commercial ELISA kits (eBioscience, Austria). The mean number of trophozoites in Ato + Astra group was found significantly lower than the number of trophozoites in the other three groups (p< 0.05). The number of trophozoites in the atovaquone and astragalus groups were found significantly lower than the number of trophozoites in the control group (p< 0.05). There was a significant increase in IL-2 levels of astragalus group compared with the other three groups, in addition when IL-2 levels of Ato + Astra group were compared with ones in other three groups, a significant decrease was noticed (p< 0.05). There was a definite increase in IL-12 levels of atovaquone, astragalus and the control groups compared to those in Ato + Astra group (p< 0.05). A significant increase was found in IFN-γ levels in atovaquone and Ato + Astra groups compared with those in the control group (p< 0.05). Within the reach of our literature survey, this study was the first research in which the effectiveness of the combination of atovaquone and AmE was investigated in the treatment of acute toxoplasmosis. The results of our study suggested that there might be a synergy between atovaquone and AmE in the treatment of acute toxoplasmosis. In case these results are supported by further studies, atovaquone and AmE combination may have a potential to be used for therapy in immunocompromized patients such as AIDS patients who have a risk for toxoplasmosis.

Astragalus spp. and Panax spp. have a long history of traditional human use. A blend, InnoSlim®, of highly purified and fractionated root extracts from Astragalus membranaceus and Panax notoginseng has now been developed for human consumption; however, the unique constituent content of this blend has not been specifically evaluated with respect to safety. Therefore, the toxicological potential of the blend was formally investigated in a series of studies-genetic toxicity was evaluated in a bacterial reverse mutation test followed by an in vivo mammalian micronucleus test, and general toxicity was evaluated in a 28-day repeated-dose oral toxicity study in rats. No evidence of mutagenicity was observed in the bacterial tester strains used, and no evidence of in vivo chromosomal damage resulting in increased frequency of micronucleated cells was observed in male Crl:NMRI BR mice. No mortality or toxic effects were observed, and no target organs were identified, in male and female Han:WIST rats exposed to 0, 400, 800, or 1200 mg/kg bw/day of the blend by gavage for 28 consecutive days. The highest dose-1200 mg/kg bw/day-was determined to be the NOAEL. Based on these results, extrapolation towards a safe human consumption level can be explored.

Water-soluble polysaccharides from 51 batches of fruits of L. barbarum (wolfberry) in China were investigated and compared using saccharide mapping, partial acid hydrolysis, single and composite enzymatic digestion, followed by polysaccharide analysis by using carbohydrate gel electrophoresis (PACE) analysis and high performance thin layer chromatography (HPTLC) analysis, respectively. Results showed that multiple PACE and HPTLC fingerprints of partial acid and enzymatic hydrolysates of polysaccharides from L. barbarum in China were similar, respectively. In addition, results indicated that β-1,3-glucosidic, α-1,4-galactosiduronic and α-1,5-arabinosidic linkages existed in polysaccharides from L. barbarum collected in China, and the similarity of polysaccharides in L. barbarum collected from different regions of China was pretty high, which are helpful for the improvement of the performance of polysaccharides from L. barbarum in functional/health foods area. Furthermore, polysaccharides from Panax notoginseng, Angelica sinensis, and Astragalus membranaceus var. mongholicus were successfully distinguished from those of L. barbarum based on their PACE fingerprints. These results were beneficial to improve the quality control of polysaccharides from L. barabrum and their products, which suggested that saccharide mapping based on PACE and HPTLC analysis could be a routine approach for quality control of polysaccharides.

OBJECTIVE

To study the therapeutic effect of traditional Chinese herbal medicinal preparation Tangshenqing (TSQ) combined with alprostadil in the treatment of early- and intermediate-stage diabetic nephropathy (DN).

METHODS

One hundred and twenty DN patients were randomized into 3 groups for different treatment protocols. The patients in the control group were given the basic treatment (low-protein diabetic diet and rigorous control of blood glucose, blood pressure, and blood fatty acid), and those in treatment group A received TSQ (containing Astragalus membranaceus, Panax notoginseng, Epimedium brevicornum, etc) in addition to the basic treatment, and those in treatment group B were treated with alprostadil injections (for 14 consecutive days) in addition to the treatment given in group A. Therapeutic effect evaluation was carried out after a 30-day treatment in all the patients.

RESULTS

The overall efficaey rate of the treatment was 78.37% in the control group, 88.57% in the treatment group A, and 94.44% in treatment group B, suggesting better therapeutic effect in the latter two groups than in the control group (P<0.05). Patients in all the 3 groups exhibited symptomatic improvement of various degrees, but the treatment group B had the best results. After the treatments, the patients' blood glucose and fatty acids were lowered, without obvious difference between the 3 groups. Compared with the control group, the patients in the two treatment groups showed significant reduction in fibrinogen, 24-h urine microprotein and urine protein after the treatment (P<0.01 or 0.05).

CONCLUSIONS

Combined use of traditional Chinese herbal medicine TSQ and alprostadil injections produces definite therapeutic effect on early- to intermediate-stage DN.

Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao (A. mongholicus, family Leguminosae) is one of the most important traditional Chinese herbs because it contains lots of bioactive metabolites, which have beneficial and pharmacological effects on health. Simultaneously, it has been proved to be a salt-tolerant plant-one of the potential species to control the soil salinization. Therefore, a sensitive and specific ultra-pressure liquid chromatography coupled with tandem mass spectrometric (UPLC-MS/MS) method was developed and validated for the simultaneous determination of six main bioactive metabolites, astragaloside IV, cycloastragenol, calycosin-7-O-β-d-glucoside, calycosin, ononin and formononetin in different organs of A. mongholicus. The detection was accomplished by multiple-reaction monitoring (MRM) scanning via electrospray ionization source operating in the positive ionization mode. Calibration curves offered linear ranges of two orders of magnitude with R(2)>> 0.99. The method was fully validated for the linearity, intra-day and inter day precisions, accuracy, recovery, matrix effect and stability. Then this method was successfully applied to detect the content of major bioactive metabolites in different plant organs of A. mongholicus under salt stress. Significant variations in the content of six bioactive metabolites were observed after been processed by different levels of salinity in different part of plant. The results support for further exploration of the salt-tolerant mechanisms in A. mongholicus and its possibility as the species that control the soil salinization. Meanwhile, we established a UPLC-MS/MS assay of the trace components in seedling of A. mongholicus in this study.

The experimental model of middle cerebral artery embolism in rats was made by inserting nylon thread. It was discovered that Astragalus membranaceus decoction(both 4.26 g.kg-1 and 7.1 g.kg-1) could reduce the capillary hyperpermeability and alleviate the dyskinesia caused by cerebral ischemia in rats. Purified Hedysarum polybotrys decoction (waste substances removed with alcohol after decocting with water) (12.5 g.kg-1) could reduce the water content in brain caused by occlusion of bilateral carotids in rats. Purified Hedysarum polybotrys decoction showed a tendency to reduce the water content in brain, but without statistical significance.

Polysaccharides from different types of natural herbs have not been compared with each other to determine their differential potencies on innate immune response, such as maturation of dendritic cells (DC). In addition, the role of endocytosis of polysaccharides in DC maturation has not been explored previously. Polysaccharides isolated from Astragalus membranaceus (APS), Ganoderma lucidum (GLP) and Radix ophiopogonis (OGP) were characterized and applied in bone marrow derived DC. Compared to immature DC, three polysaccharides with immunoactivities showed elongated dendrites, decreased phagocytic abilities, phenotypic changes (CD40/MHCII/CD80/CD86) and increased level of nitric oxide (NO) in a dose dependent manner. Interestingly, blockage of NO by iNOS inhibitor slightly decreased CD40 and MHCII but not CD80/CD86 expression induced by polysaccharides, indicating that NO was partially involved in DC maturation. In addition, GLP can enter cells in a dose and time dependent manner, shown as punctate distribution in the cytoplasm. Endocytic inhibitors sodium azide and brefeldinA that were demonstrated to inhibit cellular uptake of GLP can block phenotypic maturation of DC. Taken together, these results suggested that polysaccharides from natural herbs are effective immunostimulators with variable potencies ranking as GLP>APS>OGP, and the increase of NO level as well as the increase in polysaccharide endocytosis could be the novel strategies for improved innate response and immunotherapy.

β-elemene is an effective anticancer drug extracted from Rhizoma curcumae. It is a non cytotoxic antineoplastic agent, which can obviously inhibit the proliferation of tumor cells. In this paper, we observed the proliferation inhibition and apoptosis of β-elemene and Astragaloside on human hepatoma cell HepG2 and mouse hepatoma H22 cells, and provide a reference for further proof that β-elemene and astragaloside can induce tumor cell apoptosis. The results showed that after 24 h, group astragaloside, β-elemene group and combined treatment group had inhibitory effect on the proliferation of HePG2 cells, in which the combined treatment group had the best effect and the inhibition rate reached 66.71%. The apoptosis rates of Hep G2 cells in the drug treatment group were 0.9%, 22.4% and 45.8%, respectively, and there was statistical significance in each drug group compared with the control group (P<0.05). It can be seen that Astragalus membranaceus and β-elemene have obvious inhibitory effects on the growth of liver cancer cells and their combination has synergistic effect.

Acute kidney injury is a global problem, which brings a great burden to the society and family. The component of rhubarb, Salvia miltiorrhiza, Astragalus membranaceus, and safflower (CRSAS) has been proved as an useful agent to treat acute kidney injury (AKI) patients in China.

To assess the effect of CRSAS on human renal tubular epithelial cells (HK-2) after the hypoxia/reoxygenation (H/R) and investigate the potential mechanisms.Network pharmacology was used to predict the potential pathways shared by CRSAS and AKI. Cell counting kit-8 (CCK-8) was used to assess the HK-2 vitality. Apoptosis of HK-2 cells was detected by carboxyfluorescein succinimidyl ester/propidium iodide (CFSF/PI) staining. Expression of GRP78, CHOP, caspase-3, and Bax was detected by western blot and quantitative real-time RT-PCR.CRSAS and AKI shared the endoplasmic reticulum stress (ERS) pathway based on network pharmacology analysis. CRSAS increases the vitality of HK-2 cells and reduces the apoptosis of HK-2 cells induced by H/R injury. The expression of GRP78 and CHOP in CRSAS groups was lower than that of control groups.H/R can induce HK-2 cell apoptosis and ERS. CRSAS can reduce HK-2 cell apoptosis by inhibiting the ERS. Therefore, CRSAS might be able to treat kidney disease due to I/R injury. Animal experiment should be done to further prove our finding.