Astragaloside (AST) is derived from the Chinese herb <em>Astragalus membranaceus</em>, and studies have demonstrated that it promotes differentiation of bone marrow‑derived mesenchymal stem cells (BMSCs). To the best of our knowledge, however, the functions of the component AST‑IV in osteogenesis have not previously been elucidated. The present study aimed to verify the effects of AST‑IV in osteogenesis. First, the proliferation and differentiation status of human BMSCs incubated with AST‑IV were analysed and compared with a control (no AST‑IV treatment). In order to determine the involvement of the glycogen synthase kinase (GSK)3β signalling pathway in AST‑IV, overexpression and inhibition of GSK3β was induced during incubation of BMSCs with AST‑IV. In order to investigate how neuronal growth factor (NGF) contributes to BMSCs differentiation, BMSCs were co‑incubated with an anti‑NGF antibody and AST IV, and then levels of osteogenesis markers were assessed. The results demonstrated for the first time that AST‑IV contributed to BMSCs differentiation. Furthermore, the GSK3β/β‑catenin signalling pathway was revealed to be involved in AST‑IV‑induced osteogenesis; moreover, AST‑IV accelerated differentiation by enhancing the expression levels of NGF. In summary, the present study demonstrated that AST‑IV promotes BMSCs differentiation, thus providing a potential target for the treatment of osteoporosis.

Cisplatin (CDDP)-induced acute kidney injury (AKI) limits the therapeutic use of CDDP, which urgently needs to be addressed. Our previous study demonstrated that astragaloside IV (AS IV), an active compound of the traditional Chinese herb Astragalus membranaceus, alleviated CDDP-induced AKI. To explore the mechanism, we performed a metabolomics study to explore the altered metabolic pathways and screen for sensitive biomarkers. Twenty-four rats were randomly divided into three groups, which were treated with vehicle solutions (Control), intraperitoneally injected CDDP, and intraperitoneally injected CDDP plus oral AS IV, respectively. Metabolic profiles of serum, urine, and kidney samples were analyzed by high-performance liquid chromatography-time of flight mass spectrometry. There were 38 key metabolites in the urine samples, 20 in the serum samples, and 16 in the kidney samples that were significantly altered due to AS IV-mediated protection against CDDP-induced AKI relative to CDDP-only treatment. CDDP + AS IV co-treatment significantly altered two pathways in the blood (biosynthesis of unsaturated fatty acids and alanine, aspartate, and glutamate metabolism), five pathways in the urine (phenylalanine metabolism; phenylalanine, tyrosine, and tryptophan biosynthesis; arginine biosynthesis; arginine and proline metabolism; and histidine metabolism), and five pathways in the kidneys (glutathione metabolism; alanine, aspartate, and glutamate metabolism; glyoxylate and dicarboxylate metabolism; arginine and proline metabolism; and D-glutamine and D-glutamate metabolism). The metabolic pathways were mainly associated with improvements in inflammatory responses, oxidative stress, and energy metabolism. Adrenic acid in serum and L-histidine and L-methionine in urine were identified as sensitive biomarkers. This study provides new insights to understand the mechanism of AS IV-mediated protection against CDDP-induced AKI and has identified three candidate biomarkers to evaluate preventative treatment and assess therapeutic effectiveness.

Keywords: Acute kidney injury; Astragaloside IV; Biomarkers; Cisplatin; Metabolomics.

Ethnopharmacological relevance: Astragaloside IV (AST IV) is the active component of Astragalus membranaceus (Fisch.) Bunge, which regulates lipid and carbohydrate metabolism and improves insulin resistance. In this study, we investigated the effects of AST IV on insulin resistant cells and a non-alcoholic fatty liver disease (NAFLD) model induced by high-concentration insulin or oleic acid (OA) in HepG2 cells, as well as the associated regulatory markers.

Methods: First, the target of AST IV was predicted via pharmacophore model matching and molecular docking. Then, enzyme kinetics experiments were conducted in vitro to determine the effect of AST IV on the target protein. Next, AST IV's toxicity was tested on HepG2 cells in vitro, through an insulin resistance model and an NAFLD model, by high-concentration insulin or OA, respectively. To explore the effects of AST IV on insulin resistance and lipid metabolism, we detected the related indexes of glucose and lipid metabolism through commercially available kits. Relevant proteins were also detected by western blot to provide future direction for study.

Results: Our preliminary results of pharmacophore model matching and molecular docking suggest that AST IV and protein tyrosine phosphatase 1B (PTP1B) can be well-combined through hydrogen bonding. Further, the enzyme kinetics experiment showed that AST IV was an effective and specific inhibitor to PTP1B. We found that the protein levels of PTP1B in HepG2 cells were significantly increased after treating with high-concentration insulin or OA. Additionally, the intervention of AST IV significantly increased glucose consumption in an insulin resistance model and reduced the content of triglyceride (TG), total cholesterol (TC), and free fatty acid (FFA) in the NAFLD model. Moreover, the 2-N-(7-nitrobenze-2-oxa-1, 3 diazol-4-yl) (2-NBDG) uptake rate in the NAFLD model was also greatly improved. These results validated the effects of AST IV on improving insulin resistance and lipid accumulation. Furthermore, western blot results illustrated that AST IV suppressed PTP1B and increased levels of phosphorylated insulin receptor (p-IR) and phosphorylated insulin receptor substrate-1 (p-IRS-1) in insulin-resistant HepG2 cells, while also decreasing protein levels of PTP1B and sterol element regulatory binding protein-1c (SREBP-1c) in the NAFLD model.

Conclusion: This study demonstrated that AST IV inhibited PTP1B and effectively improved insulin resistance in insulin-resistant HepG2 cells and triglyceride accumulation in OA-treated HepG2 cells.

Keywords: Astragaloside IV; Insulin resistance; Lipid metabolism; Protein tyrosine phosphatase 1B (PTP1B).

Despite minoxidil and finasteride already being approved by the Food and Drug Administration (FDA) for the treatment of hair loss, it is important to identify new and innovative treatments for hair loss, such as looking for a solution in Chinese herbal medicine. One such treatment to consider is BeauTop (BT), whose primary ingredients include Panax japonicus (T.Nees), C.A. Mey. (Araliaceae), Astragalus membranaceus (Fisch) Bunge (Fabaceae), Angelica sinensis (Oliv.) Diels (Apiaceae), Ligustrum lucidum W.T. Aiton (Oleaceae), Rehmannia glutinosa (Gaertn.) DC. (Plantaginaceae), and Eclipta prostrata (L.) L. (Compositae). The aim of this study was to evaluate whether BT can promote hair growth in C57BL/6 mice and to investigate hair coverage, the expression of vascular endothelial growth factor (VEFG), and the numbers of hair follicles in growth phase after oral administration. A total of 12 C57BL/6 mice were divided into two groups: control group and treatment group BT. BT was administered orally as an extract at a volume of 0.6 g/kg. The control group was treated with distilled water. Each group was treated once a day for 12 consecutive days. To observe the expression of VEGF distribution, the number of hair follicles and the hair coverage were examined on days 4, 8, and 12. By comparing the treatment group and control group, we found that VEGF in the BT group on day 8 presented with a higher area percentage than the control group (p value = 0.003). Hair follicle counting results showed that the BT group was significantly higher than the control group on day 8 (p value = 0.031). Furthermore, hair coverage was shown to be significantly increased in the treatment group BT on day 8 (p value = 0.013). Taken together, these results suggest that Chinese medicine (BT) possesses the potential effect of promoting hair growth through VEGF expression. VEGF is considered the most important mediator for the process of angiogenesis involved in hair growth development.

This paper deals with the prospective clinical study on treatment of acute upper digestive tract hemorrhage with Wen-She decoction (WSD). An opened sequential controlled trial method of simple orientation quality reaction was adopted in this study. 7 cases were treated and all of them were cured. It was concluded that WSD was an excellent therapy to treat the middle or small amount hemorrhage of acute upper digestive tract. The effective rate of WSD of the stool OB (+) becoming (-) within 5 days was more than 95%. WSD consists of Codonopsis pilosulae, Atractylodes macrocephala, Poria cocos, Glycyrrhiza uralensis, Zingiber officinale, Os sepiae Halloysitum rubrum and Astragalus membranaceus. It is effective in stopping bleeding by warming the Spleen and tonifying Qi.

Fatty acids are important nutritional substances and metabolites in living organisms. These acids are abundant in Chinese herbs, such as Brucea javanica, Notopterygium forbesii, Isatis tinctoria, Astragalus membranaceus, and Aconitum szechenyianum. This review illustrates the types of fatty acids and their significant roles in the human body. Many analytical methods are used for the qualitative and quantitative evaluation of fatty acids. Some of the methods used to analyze fatty acids in more than 30 kinds of plants, drugs, and other samples are presented in this paper. These analytical methods include gas chromatography, liquid chromatography, near-infrared spectroscopy, and NMR spectroscopy. The advantages and disadvantages of these techniques are described and compared. This review provides a valuable reference for establishing methods for fatty acid determination.

One hundred and forty-four fungal isolates were obtained from diseased Paeonia albiflora Pall. var. trichocarpa Bung., Astragalus membranaceus Bung., Lithospermum erythrorhizon Sieb. et Zucc., Ledebouriella seseloides Wolff and Bupleurum falcatum L. which were collected in the test field of Tsukuba Medicinal Plant Research Station, National Institute of Hygienic Sciences. Most of them were identified into 15 genera containing 8 species. Fungal species presumed to be pathogens of the host plants were as follows: Cladosporium paeoniae, Pestalotia paeoniicola, Glomerella cingulata, Hainesia lythri, Guignardia sp. and Alternaria sp. from P. albiflora, Fusarium spp., Rhizoctonia spp. and Neocosmospora vasinfecta from A. membranaceus, Colletotrichum gloeosporioides from L. erythrorhizon, Rhizoctonia sp., Fusarium spp., Phoma sp. and Pyrenochaeta sp. from L. seseloides, and Fusarium sp., Alternaria alternata, Phyllosticta sp., Phoma sp., Phomopsis sp. and C. gloeosporioides from B. falcatum. Roots of B. falcatum were found to be parasitized by Meloidogyne sp.

OBJECTIVE

To observe the effect of Compound Fructus Arctii Mixture (FAM, consisted of Fructus Arctii and ethanol extract of Radix Astragalus membranaceus) in treating diabetic nephropathy.

METHODS

Using FAM to treat 31 patients with diabetic nephropathy and controlled by 23 patients treated with Losartan, the therapeutic course was 3 months for both groups, changes of clinical symptoms, blood glucose, lipid metabolism and urinary albumin were observed and compared.

RESULTS

The total effective rate in the treated group was 80.6% while in the control group 65.2%. The symptoms, urinary protein and albumin as well as lipid metabolism in the treated group all significantly improved after treatment (P<0.05), but in the control group improvement only showed in urinary albumin level (P<0.05).

CONCLUSIONS

FAM has the effects of reducing urinary protein in 24 hrs, lowering urinary albumin, improving blood glucose after meal and lipid metabolism.

OBJECTIVE

To clarify the effects of Astragalus Membranaceus (AM) combined with taurine and/or coenzyme Q10(CoQ10) on coxsackievirus B3 (CVB3) murine myocarditis.

METHODS

Viral myocarditis model was created by intraperitoneal inoculation with CVB3 solution and were treated by saline, AM, taurine, CoQ10, AM + taurine, AM + CoQ10, AM + taurine + CoQ10, respectively. The mortality, ECG, CVB3-RNA in myocardium and myocardial histopathologic changes were observed.

RESULTS

AM combined with taurine and CoQ10 could significantly reduce the mortality of the mice and the incidence of abnormal ECG at acute stage. CVB3-RNA was significantly reduced in AM treated group, especially in AM + taurine group. No anti-virus effect was found in CoQ10 group. All drugs could lighten myocardial histopathologic changes and the effect could be enhanced by combined treatment.

CONCLUSIONS

AM, taurine and CoQ10 have some curative effects on CVB3 murine myocarditis, AM combined with taurine and CoQ10 is the best.

Large dose of Yang-restoring herb medicines. (Radix Codonopsis Pilosulae, Astragalus membranaceus, Radix Aconiti Praeparata, Epimedium brevicornum, Cortex Cinnamomi and Herba Cistanchis) may exert an unfavorable effect on normal rats, i. e. natural weight gain reduced (P less than 0.01), serum T3 decreased (P less than 0.05), rT3, TRH levels raised (P less than 0.01) and TSH showed a raising tendency. Lower T3 and higher rT3 levels may be the effects of Yang-restoring herb medicines on the peripheral metabolism of thyroid hormones, i. e. more thyroxine was degraded to rT3 with little biological effect and less was transformed to T3 with strong hormonal effect. This unfavorable effect, however, can be avoided by Forward-Backward method. It was advised that large dose of Yang-restoring herb medicines could not be given to the organisms without symptoms of Yang-deficiency. If it were tried to do so, Forward-Backward method might be recommended.

Infantile pneumonia has been treated successfully with Huayu decoction (HYD) in our department for more than ten years. According to the principles of Huoxue Huayu of TCM, the composition of Huayu decoction is as follows: Angelica sinensis, Paeonia lactiflora, Ligusticum wallichii, Spathollobus suberectus, Hirudo nipponica, Tabanus bivittatus, Paeonia suffroticosa, Astragalus membranaceus . 49 cases of infantile pneumonia were treated with HYD. The activity of erythrocyte superoxide dismutases (ESOD) was measured in these patients. The results showed that the activity of ESOD reduced in the acute stage and returned to normal in convalescence. There was significant statistical difference in activity of ESOD between the patient's group. In experimental studies, the producing of free radicals was induced by inhaling ozone in mice. It was found that HYD had the effect of scavenging free radicals in these animal models. The action of anti-oxidate of HYD was also detected in vitro. The mechanism of HYD in treating infantile pneumonia might be elucidated in some respects by these clinical and experimental studies.

OBJECTIVE

To investigate and explain the composing mechanism by observing enhancement property of disassembled compositions of Yiqi Huoxue decoction (YQHXD) on platelet aggregation of health adults.

METHODS

Venous whole blood was obtained from healthy human subjects and anti-coagulated, and then centrifuged to produce platelet-rich plasma (PRP) and platelet-poor plasma (PPP). Disassembled compositions of YQHXD and sodium chloride were placed in different translucent tubes containing PRP and PPP, calculated the maximum percents of platelet aggregation stimulated by platelet agonists: adenosine diphosphate (ADP), platelet activating factor (PAF) and arachidonic acid (AA).

RESULTS

The maximum percents of platelet aggregation induced by ADP and PAF were largely improved by YQHXD, both astragalus membranaceus and Angelica Scinesis played the most important roles in the decoction, the herbs of Yiqi and Huoxue must work together to enhance platelet aggregation.

CONCLUSIONS

In vitro study, YQHXD can markedly improve platelet aggregation of health adults, but the exact mechanism should be proved by further experiments.

Kang Er Xin-I (KEX-I) is a proved recipe used to treat viral myocarditis. It consists of Lonicera japonica, Ophiopogon japonicus, Astragalus membranaceus mainly and possesses the effect of clearing away heat and toxic materials and supplementing the vital energy and nourishing. The clinical study was carried out with KEX-I according to a random, paired and cross-over design. Coenzyme Q10 was used as a control and left ventricular function was observed. The result showed: after being treated with KEX-I for two weeks, the 26 patients' chief cardiac functional indexes assessed with STI improved markedly, the value of PEP/LVET and ICT/LVCT all decreased and the difference between the two groups was significant. The experimental study showed that KEX-I can inactivate directly the virus of Coxsackie B3, protect the heart cells in mice, prevent attack by Coxsackie B3, promote the growth of internal interferon and increase the NK cell's function to regulate immunity in experimental mice.

OBJECTIVE

To study difference among populations which belong to Astragalus membranaceus and A. membranaceus var. mongholicus on morphology, habit, characteristics of physiology and resistance to powdery mildew, and classify them in order to provide theoretical basis for breeding and improving varieties.

METHODS

Morphology, habits, isozyme and soluble protein electrophoretograms were compared among the populations. They were categorized by cluster analysis based on those electrophoretograms. Different ability of resistance to powdery mildew was also studied through comparing disease indices among six populations.

RESULTS

The results showed A. membranaceus var. mongholicus was distinctly different from A. membranaceus. There was a special type in colonies of A. membranaceus, which showed hairy upper epidermis of leaflets and later florescence.

CONCLUSIONS

Astragalus for medicine could be categorized in three types A. membranaceus var. mongholicus, A. membranaceus early florescence type and A. membranaceus late florescence type. Among them A. membranaceus var. mongholicus is most resistant to powdery mildew, while A. membranaceus is easily infected, and the early florescence type is even more easily infected.

Chinese herbal medicine and gentamycin were used separately in the treatment of 75 cases of gastric disease by campylobacter pyloridis (CP). Surveying the changes of the gastrofiberscope, pathological test, bacteriology and immunology. This study, found that in the Chinese herbal medicine group clinical symptoms were obviously improving the effective rate for CP being 80% (24/30), the death rate for CP 30% (9/30), and the effective rate for patho-histological changes 50% (15/30). Differences of curative effective rate between the two groups were statistically insignificant (P greater than 0.05). This text pointed out that the principle of Chinese herbal medicine treatment of gastric disease by campylobacter pyloridis was fu-zheng qu-xie. Fu-zheng was achieved by Astragalus membranaceus, Atractylodes macrocephala and Paeonia lactiflora, whereas qu-xie by Taraxacum monogolicum and Oldenlandia diffusa. Chinese herbal medicine for fu-zheng played an important role in modulating immune function. Qu-xie was directly disinfective and indirectly anti-bacterial. Chinese herbal medicine combined with western drugs will decrease the side effects and enhance the curative effect at the same time.

Effects of water and ethanol extracts of 10 Chinese medicines, such as Astragalus membranaceus, Coix lachryma-jobi, etc., on biomass and exopolysaccharide of Ganoderma lucidum were studied by submerged culture. The results showed: water extracts of all medicines can improve the culture of G. lucidum except of A. membranaceus, ethanol extracts of C. lachryma-jobi, Dioscorea opposita, Codonopsis pilosula, and Achyranthes bidentata( < 187.5g Medicine/L substrate) can also increase the biomass of G. lucidum, but the ethanol extracts of Angelica sinensis, Dendrobium nobile check the growth of G. lucidum. The production of exopolysa-ccharide can be improved by all the Chinese medicines and their dosage used in this experiment, Although A. sinensis, D. Nobile check the growth of G. lucidum, they could stimulate the secretion of exopolysaccharide in lower dosage. It is concluded that some Chinese medicines, such as C. lachryma-jobi, D. opposita, C. pilosula, etc. can be processed by the fermentation of G. lucidum, and bio-active compound can be produced by adding appropriate Chinese medicine in the substrate to culture G. lucidum.

A new agar-free bioautographic assay for xanthine oxidase (XO) inhibitors and superoxide scavengers on TLC layers was developed and validated. Compared to the first version of TLC bioautographic agar overlay method, our bioautographic assay greatly improved the sensitivity and quantification ability. The limit of detection (LOD) of this assay was 0.017ng for allopurinol. Quantitative estimation of XO inhibitors and superoxide scavengers was achieved by densitometry scanning, expressed as allopurinol equivalents in millimoles on a per sample weight basis. This assay has acceptable accuracy (95.37-99.23%), intra-day and inter-day precisions (RSD, 2.56-6.69%), as well as intra-plate and inter-plate precisions (RSD, 2.93-9.62%). Six pure compounds and three herbal extracts were evaluated for their potential XO inhibitory and superoxide scavenging activity by this bioautographic assay on TLC layers. Four active components were separated, located and identified in Astragalus membranaceus var. mongholicus extract by the bioautographic assay after TLC separation. The developed method is rapid, simple, sensitive and stable for screening and estimation of the potential XO inhibitors and superoxide scavengers.

The polysaccharides of different germplasm resources of Astragalus membranaceus var. mongholicus〓(cultured Astragalus Radix (RA) and natural RA) and A. membranaceus (MJ) (cultured RA and natural RA) were studied by using the optimal enzymatic conditions of endo-1,4-β-mannanase. Saccharide fingerprints were obtained for the identification and evaluation of the germplasm resources of RA by Fluorophore-assisted Carbohydrate Electrophoresis (FACE). The data were analyzed by principal component analysis to obtain the difference between RA of different germplasm resources. The results showed that trisaccharide, tetrasaccharide and pentasaccharide of endo-1,4-β-mannanase hydrolyzate could be used as the differential fragments to distinguish MG (cultured RA and natural RA); the pentasaccharide and hexasaccharide can be used as differentially expressed carbohydrate fragments that distinguish MJ (cultured RA and natural RA); the trisaccharide and tetrasaccharide can be used as the differential fragments to distinguish the cultured MG and cultured MJ. Studies have shown that polysaccharide products degraded by endo-1,4-β-mannanase can well distinguish RA species (MG and MJ), growth mode (cultured RA and natural RA). This study laid the foundation for the quality evaluation of Astragalus medicinal herbs and screening of active oligosaccharides.

To identify Oxytropis medicinal materials using ITS2 sequence.

The second internal transcribed spacer( ITS2) of Oxytropis fetissovii, Oxytropis myriophylla and Oxytropis grandiflora medicinal material samples was amplified by PCR and sequenced. To expand scope of the research topic,ITS2 sequences of related species were downloaded from Gen Bank. Sequences assembly and consensus sequence generation were performed by ITS2 Database. The related data analysis and processing was performed using software MEGA 5. 10 and the NJ tree was constructed. The ITS2 secondary structure was predicted using ITS2 web server, and the differences of the ITS2 secondary structures of the samples were analyzed.

Oxytropis medicinal materials ITS2 sequence was shorter, with sequence length of 216 ~ 218 bp, which was in favor of DNA extraction, PCR amplification and sequencing. Genetic distances of Oxytropis myriophylla, Oxytropis fetissovii and Oxytropis grandiflora were much larger than the genetic distances of themselves. In the NJ tree, Oxytropis medicinal materials and counterfeits could be distinguished, and Oxytropis medicinal materials could be distinguished from Astragalus membranaceus.

The DNA barcode based on ITS2 sequence is a powerful and efficient tool for identification of Oxytropis medicinal materials.

PG2 is an infusible polysaccharide extracted from Astragalus membranaceus, which is a Chinese herb traditionally used for stroke treatment. We investigated the effect of PG2 on patients with spontaneous acute intracerebral hemorrhage (ICH). A total of 61 patients with acute spontaneous ICH were randomized to either the treatment group (TG, 30 patients), which received 3 doses of PG2 (500 mg, IV) per week for 2 weeks, or the control group (CG, 31 patients), which received PG2 placebo. At 84 days after PG2 administration, the percentage of patients with a good Glasgow outcome scale (GOS 4-5) score in the TG was similar to that in the CG (69.0% vs. 48.4%; p = 0.2). The percentage of good mRS scores (0-2) in the TG was similar to that in the CG (62.1% vs. 45.2%; p = 0.3). In addition, no significant differences were seen when comparing differences in the C-reactive protein, erythrocyte sedimentation rate, interleukin-6 (IL-6), IL-1β, tumor necrosis factor-α, and S100B levels between baseline and days 4, 7, and 14 after PG2 administration (all p>> 0.05). The results are preliminary, necessitating a more thorough assessment.