An experiment was conducted to investigate the effects of polysaccharide extracts (E) of two mushrooms, Lentinus edodes (LenE) and Tremella fuciformis (TreE), and an herb, Astragalus membranaceus (AstE), on the immune responses of chickens infected with Eimeria tenella. A total of 180 broiler chickens were assigned to nine groups: three groups were fed with each of the extracts (LenE, TreE, and AstE), three groups were fed with the extracts and immunized with live oocyst vaccine (LenE+V, TreE+V, and AstE+V), a group was immunized with the vaccine only, and there were two controls (E. tenella-infected and noninfected groups). The oocyst vaccine was given at 4 days of age, and the extracts (1 g/kg of the diet) were supplemented from 8 to 14 days of age. At 18 days of age, all birds except those in the noninfected group were infected with 9 x 10(4) sporulated oocysts. The results showed that at 7 days postinfection (p.i.), birds fed the extracts without vaccination had lower body weight (BW) gain than those given the vaccine only. However, the extracts in conjunction with the vaccine significantly enhanced BW gain of the infected chickens compared with the vaccine group. Of the three extracts, LenE and TreE showed a better growth-promoting effect. The extracts largely increased oocyst excretion of droppings during the primary response postvaccination. The cecal peak oocyst output and lesion scores measured at 7 days p.i. were higher in the groups fed the extracts than in the group immunized with the vaccine only, whereas those of the groups fed with the extracts and immunized with the vaccine were not significantly different from the vaccine group. Of the three extracts, both LenE- and AstE-fed groups showed lower cecal oocyst output. Thus, as compared with the extracts, the live, attenuated vaccine showed better results with significantly increased immune response in coccidial infected birds. The polysaccharide extracts may prove useful against avian coccidiosis, and, particularly when they are used in conjunction with vaccine, they have shown preliminary promise against the experimental coccidial infection.

Astragalus membranaceus (Fisch.) Bunge, a commonly used Chinese medicinal material, from certain localities contains more favourable trace elements and fewer harmful trace elements than those from other localities. Therefore, there is a need to distinguish Astragalus membranaceus from different localities. Internal transcribed spacer 1 (ITS1) of the nuclear ribosomal RNA gene of 23 Astragalus membranaceus samples were sequenced to confirm the species of the samples. Arbitrarily primed polymerase chain reaction (APPCR) was then used to obtain unique fingerprints for each sample using several primers. The presence and absence of bands were used for calculating mean similarity indexes among the samples. It was found that the Heilongjiang samples were markedly distinguishable from samples of other localities. In addition, bands common for samples from the same locality were also identified and used to distinguish samples from Neimengu and Shanxi. Therefore, Astragalus membranaceus from these provinces, the major cultivation places in China, can be differentiated.

It has been known that the renal microvasular lesions could aggravate the progress of glomerular sclerosis and tubulo-interstitial fibrosis in chronic kidney diseases. Modern pharmacological studies indicated that the two traditional Chinese herbs, Astragalus membranaceus and Angelica sinensis, could improve micorvascular lesions through multiple mechanisms, including increasing local renal blood flow to lessen the hypoxic renal injury, promoting the recovery of renal blood flow and glomerular filtration rate after ischemia-reperfusion, modulating the imbalance of vaso-activators such as nitric oxide and angiotensin, increasing the expression of vascular epithelial growth factor and inhibiting the release of the intracellular calcium ion and promoting DNA synthesis in endothelial cells to improve the function of endothelial cells. These evidences suggest that Astragalus membranaceus and Angelica sinensis may retard the progress of renal diseases through the above-mentioned mechanisms.

OBJECTIVE

To explore the angiogenesis effect of Astragalus membranaceus and Angelica sinensis with different ratio in chick embryo chorioallantoic membrane (CAM).

METHODS

Chicken CAM model was adopted. The rat blood serum containing different ratio of Astragalus membranaceus and Angelica sinensis was put daily into 3-day chick embryo. CAMs and CAM angiogenesis were observed at 72 h after incubation.

RESULTS

Better effects were obtained in the formation of capillary and the count of blood vessel in groups treated by Astragalus memhranaceus and Angelica sinensis in the ratio of five to one, compared with the NS control group and the other ratio groups of Astragalus memhranaceus and Angelica sinensis.

CONCLUSIONS

Astragalus memhranaceus and Angelica sinensis in 5:1 ratio have certain promotive effect on the formation of capillary. The Danggui Buxue Decoction has the best ratio.

Many Chinese herbs and herbal mixtures are fed to domestic animals for their reputed medicinal properties. These herbs could contribute to the intake of essential nutrients and toxic metals, but their composition is mostly unknown. The purpose of this study was to measure major nutrient (crude protein, crude fat, carbohydrate, fibre) and mineral (Ca, P, Mg, K, Na, Fe, Zn, Cu, Mn, Mo, S, Al, Cd, Ni, Pb) concentrations in samples of fourteen combination formulas labelled for veterinary use and commonly administered to horses and dogs. Three single herbs, Bupleurum chinense, Curcuma zedoaria and Astragalus membranaceus, each obtained from several sources, and Yunnan Baiyao, a proprietary hemostatic mixture, were also analysed. Proximate analyses and some mineral concentrations differed (p < 0.05) among single herbs, and high concentrations of several minerals were detected in some herbal combinations. Those containing the highest concentrations [g/kg dry matter (DM)] of calcium (92.4), iron (2.6) and manganese (0.28) could provide >38%, 142% and 96%, respectively, of recommended allowances in adult dogs, and >13%, 122% and 2%, respectively, of maintenance requirements in horses, at the maximum labelled dose assuming complete availability. Concentrations of cadmium, nickel and lead were below published oral tolerance levels. Aluminium concentrations (median 380, maximum 920 mg/kg DM) were higher than has been previously reported in Chinese herbs. These nutrient analyses suggest that herbal combinations marketed to veterinarians, when fed at the maximal labelled dose, are unlikely to produce clinically relevant changes in the dietary intake of essential nutrients. However, small amounts of non-essential contaminant minerals are present in some formulas, and further research is necessary to understand the significance of this finding.

On the basis of outpatients' medical records concerning primary liver cancer (PLC), data of 552 patients (with 2020 effective prescriptions) from the Outpatient Department of Changhai Hospital treated by Professor Ling Changquan were collected. The nature, flavor and meridian distribution of the herbs used in the prescriptions were summarized by frequency method, and the features of the herbs used according to syndrome differentiation were analyzed by logistic regression. The couple herbs used were analyzed by cluster analysis. All the data were analyzed in combination with the experience of the specialist. It showed that most of the frequently used herbs were herbs for invigorating the spleen to promote appetite, removing toxic materials to inhibit tumor growth, and activating blood circulation to dissipate blood stasis. These herbs are mostly of plain or cool nature, and mainly of sweet, bitter, or acrid taste. It also showed that the most frequently used herbs for qi deficiency were Astragalus membranaceus, Atractylodes macrocephala, and Pseudostellariae; Caulis Spatholobi and Chinese jujube for blood asthenia; Fructus Corni and Ophiopogon japonicus for yin deficiency; Agastache rugosa, Semen Plantaginis and Poria for water-dampness; cape jasmine fruit and baikal skullcap root for heat excess; peach seed and Radix Paeoniae Rubra for blood stasis; Curcuma wenyujin, Akebia trifoliata and Bupleurum chinese for qi stagnation. A total of 19 pairs of couple herbs were summarized by the cluster analysis.

Polysaccharides isolated from traditional Chinese medicines (TCMs) exhibit multiple pharmacological activities. However, quality control of polysaccharides is a challenge because of their complicate structure and macro-molecular mass. In this study, saccharide mapping based on specific enzymatic digestion of polysaccharides and chromatographic analysis was proposed to discriminate the polysaccharides from different TCMs. Endo-carbohydrase such as glucanase, arabinanase, xylanase, galactanase, cellulase, amylase and pectinase were used for enzymatic digestion of polysaccharides from 9 TCMs namely Panax ginseng, P. notoginseng, P. quinquefolium, Cordyceps sinensis, C. militaris, Ganoderma lucidum, G. sinense, Astragalus membranaceus and Angelica sinensis. By using high performance size exclusion chromatography (HPSEC) as well as derivatization with 1-Phenyl-3-methyl-5-pyrazolone (PMP) and HPLC analysis, the enzymatic hydrolysis properties of polysaccharides and their saccharide mapping were determined. The polysaccharides from 9 TCMs were firstly successfully distinguished based on their characteristic saccharide maps, which is helpful to improve the quality control of polysaccharides.

Tetracyclic triterpenoids, including the dammarane, cucurbitane, cycloartane, lanostane and protostane groups, is a class of triterpenoids widely distributed in various medicinal plants, particularly those commonly used for the treatment of diabetes and its complications, such as Panax ginseng, Panax quinquefolium, Panax notoginseng, Gynostemma pentaphyllum, Astragalus membranaceus, Momordica charantia, and Ganoderma lucidum. This review highlights recent findings on the chemistry and bioactivities of tetracyclic triterpenoids from these plants and other popular herbal medicines.

OBJECTIVE

To investigate whether Astragalus membranaceus (AM) can protect endothelium-dependent vasodilatation (EDV) function of aorta from the damage induced by high level of free fatty acid (FFA).

METHODS

Ten male SD rats, 8 weeks old and 250-300 g in weight, were sacrificed and thoracic aorta were harvested. Aorta rings incubated in organ baths were divided into three groups, Control group, FFA group and FFA+ AM group. The control group was incubated in 20 mL Krebs-Henseleit solution; the FFA group was incubated in 20 mL KH solution mixed with FFA(800 micromol/L) the FFA + AM group was incubated in 20 mL KH solution mixed with FFA (800 micromol/L) and AM (4 g/L). The relaxation levels of aorta rings response to acetylcholine and sodium nitroprusside were measured, the expression of NF-kappaB and the level of NOx in the organ bath were analyzed by immunohistochemistry.

RESULTS

Severe endothelial dysfunction were induced in FFA group (maximal vasorelaxation in response to Ach: 61.1% +/- l6.9% vs. 93.1% +/- 2.7% in control, P < 0.05), while EDV in FFA+AM group was significantly improved by the incubation with AM (P < 0.05). Compared with the control group (104.1 +/- 14.2) micromol/g, NOx levels of FFA group was (83.1 +/- 8.4) micromol/g (P < 0.05), and the treatment of AM increased the levels of NOx (98.8 +/- 10.7) micromol/g (P < 0.05). The control vascular ring had a little NF-kappaB expression in endothelial nucleus, FFA increased the activation of NF-kappaB, while the treatment of AM lower the elevated NF-kappaB level.

CONCLUSIONS

FFA can directly injure EDV, while AM may ameliorate it, with the possible mechanism related to the signal pathway of NF-kappaB and NO.

Astragaloside IV (AST) as the main active ingredient of Astragalus membranaceus. Clinical and laboratory-based studies have demonstrated the effects of AST on cerebral protection and angiogenesis after ischemia stroke. In addition, several reports investigated the effect of AST on proliferation of neural stem cells. The current study was aimed to evaluate the influence of AST on neurogenesis in hippocampal dentate gyrus (DG) of MCAO rats and to explore the possible mechanisms. In this study, the neurobehavioral tests (Ludmila Belayev 12-point scoring, Screen test, fore limb placing test) had been employed to investigate the effect of AST treatment against functional deficit of MCAO rats. The immunofluorescence staining, western-blot and qRT-PCR was performed to evaluate the effects of AST on proliferation, differentiation and maturity of neural stemr cells in hippocampus. Moreover, we investigated the possible mechanism of the AST treatment in promoting neurogenesis after ischemic stroke. The findings indicated that AST treatment ameliorated the neurobehavior of MCAO rats. The results indicated that AST treatment possessed the potential to improve proprioceptive sense and motor function of MCAO rats. AST treatment sustained neuronal viability and stimulates sensorimotor integration functional recovery in MCAO rats. The results suggested that AST improved neurobehavior deficit after ischemic stroke. Furthermore, AST promoted neurogenesis through upregulating the expressing of BNDF/TrkB signaling pathway. Therefore AST might be a promising therapeutic agent for ischemic stroke.

Keywords: Astragaloside IV; Hippocampus; Ischemic stroke; Neurogenesis.

Extracorporeal shock wave lithotripsy (ESWL)-induced renal damage can occur as a result of multiple mechanisms. We have reported previously that Astragalus membranaceus, Salvia miltiorrhiza, a decoction of six drugs containing rhizoma Rehmanniae preparata and supplements of a few traditional Chinese medicinal herbs for invigorating the kidney and excreting calculus, have a protective effect on renal injury induced by high-energy shock waves (HESW) in rabbits. In this clinical study we further investigate the protective effects of these traditional Chinese herbs against renal damage induced by ESWL. Sixty consenting patients with renal calculus who underwent ESWL treatment were included and randomly assigned to the medication group or control group. Post-ESWL plasma nitric oxide (NO), endothelin-1 (ET-1), malondialdehyde (MDA), and serum tumor necrosis factor α (TNF-α) increased significantly in the controls (P < 0.05), while in the medication group, slightly but not significantly elevated levels of plasma ET-1, NO, and serum TNF-α were found. The difference between the groups was statistically significant (P < 0.05). The levels of superoxide dismutase (SOD) decreased gradually in the controls, reaching a trough 72 h after ESWL (P < 0.05), while in the treated group it was unchanged, and remained at a level higher versus the controls (P < 0.05). Plasma NO peaked twice by 72 h and at 1 week in the controls (P < 0.05). Urinary enzymes and β(2)-microglobulin increased significantly and peaked by 24 h and immediately after ESWL (P < 0.05). These values were greater in the controls, and the difference was statistically significant (P < 0.05). This study demonstrates that the preparations of traditional Chinese medicines for invigorating the kidney and excreting calculus can reduce renal tubular damage induced by ESWL, and can shorten the recovery time of renal tubules in human subjects.

BACKGROUND

Atherosclerosis is considered to be a chronic inflammatory disease. Astragali Radix extract (ARE) is one of the major active ingredients extracted from the root of Astragalus membranaceus Bge. Although ARE has an anti-inflammatory function, its anti-atherosclerotic effects and mechanisms have not yet been elucidated.

METHODS

Murine endothelial SVEC4-10 cells were pretreated with different doses of ARE at different times prior to induction with tumor necrosis factor (TNF)-α. Cell adhesion assays were performed using THP-1 cells and assessed by enzyme-linked immunosorbent assay, western blotting and immunofluorescence analyses to detect the expression of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), phosphorylated inhibitor of κB (p-iκB) and nuclear factor (NF)-κB. We also examined the effect of ARE on atherosclerosis in the aortic endothelium of apolipoprotein E-deficient (apoE(-/-)) mice.

RESULTS

TNF-α strongly increased the expression of VCAM-1 and ICAM-1 accompanied by increased expression of p-iκB and NF-κB proteins. However, the expression levels of VCAM-1 and ICAM-1 were reduced by ARE in dose- and time-dependent manners, with the strongest effect at a dose of 120 μg/ml incubated for 4 h. This was accompanied by significantly decreased expression of p-iκB and inhibited activation of NF-κB. Immunofluorescence analysis also revealed that oral administration of ARE resulted in downregulation of adhesion molecules and decreased expression of macrophages in the aortic endothelium of apoE(-/-) mice. ARE could suppress the inflammatory reaction and inhibit the progression of atherosclerotic lesions in apoE(-/-) mice.

CONCLUSIONS

This study demonstrated that ARE might be an effective anti-inflammatory agent for the treatment of atherosclerosis, possibly acting via the decreased expression of adhesion molecules.

Background: Huangqi is a famous Chinese medicinal material whose Dao-di producing area is Hunyuan, Shanxi. Huangqi produced in Hunyuan, Shanxi, were divided into several different specifications and grades according to the diameters and different positions of root system. Objective: This article investigates the quantitative characteristics of chemical compositions in different specifications and grades of Astragalus membranaceus var. mongholicus roots, aiming to elucidate the correlation between specifications and/or grades and chemical compositions in Huangqi. Methods: Based on the field investigation, samples of Huangqi collected from Hunyuan, Shanxi, were divided into different specifications and grades. The content of seven flavonoids and five saponins in Astragalus membranaceus var. mongholicus roots of different specifications and grades were determined simultaneously by HPLC-diode-array detection-evaporative light-scattering detection (HPLC-DAD-ELSD). Results: Huangqi was processed by traditional methods, and its commercial specification was classified by different parts of the root system, such as ge-da-tou, hong-lan-qi, zheng-bai-qi, fu-bai-qi, mao-wei-zi, and qi-jian. The total content of seven flavonoids and five saponins in ge-da-tou, qi-jian were lower. The total content of seven flavonoids in hong-lan-qi was much higher, while that of five saponins was much lower. The total content of seven flavonoids in lateral roots or fibrous roots were higher, and that of five saponins was lower, such as zheng-bai-qi, fu-bai-qi, and mao-wei-zi. According to the root diameters, Huangqi was classified to special grade, grade I, grade II, grade III, grade IV, or grade V. Among six grades of Huangqi, the total content of seven flavonoids in grade III, grade IV, and grade V were lower, while the total content of five saponins in them were much higher. Conclusions: There is an obvious difference on the distribution pattern of contents of seven flavonoids and five saponins in Huangqi of different specifications and grades, which provide a certain scientific basis for the quality evaluation of Huangqi. Highlights: The content of seven flavonoids and five saponins in Huangqi were determined by HPLC-DAD-ELSD. The relationship between the commercial specification grades and chemical components of Dao-di herbs Astragalus membranaceus var. mongholicus (Huangqi ) from Hunyuan, Shanxi were revealed, which provided a chemical basis for the classification of commercial specification grades of dao-herbs Astragalus membranaceus var. mongholicus (Huangqi ) from Hunyuan, Shanxi.

Autophagy is an evolutionarily conserved 'self-eating' process that maintains cellular, tissue, and organismal homeostasis. New studies on autophagy, mediated by subsets of autophagy proteins, are emerging in many physiological and pathological processes. Astragalus membranaceus (AM), also named Huangqi, is one of the fundamental herbs in traditional Chinese medicine and its extracts have been proved to possess many biological activities related to autophagy, including anti-oxidation, anti-inflammation, anticancer, anti-photoaging, and improvement of cardiomyocyte function. Evidence suggests that AM extracts can have therapeutic potential in autophagy dysregulation-associated diseases because of their biological positive effects. Here we will review the literature concerning the effects of AM extracts on autophagy dysregulation-associated diseases.

A 12-week trial with 120 [(Landrace×Yorkshire)×Duroc] pigs (45.65 ± 1.93 kg) was conducted to evaluate the effects of Astragalus membranaceus, Codonopsis pilosula and allicin mixture (HM) supplementation on growth performance, nutrient digestibility, faecal microbial shedding, immune response and meat quality in finishing pigs. Pigs were allocated to one of three treatments with 0, 0.025% (HM1) and 0.05% (HM2) HM supplementation in a randomized complete block design according to sex and BW. Each treatment contained 10 replications with four pigs (two barrows and two gilts) per pen. Dietary HM resulted in a higher G:F (p < 0.05) than CON group during weeks 7 to 12 and the overall periods. Pigs fed HM2 diet had higher ADG than pigs fed CON diet. Pigs fed HM2 supplementation diet led to a higher (p < 0.05) apparent total tract digestibility (ATTD) of dry matter (DM) and gross energy (GE) than pigs fed CON diet at week 6, while the supplementation of HM led to a higher (p < 0.05) ATTD of DM and GE than pigs fed CON diet at week 12. The faecal E. coli counts were reduced, and Lactobacillus counts were increased by increasing HM supplementation (p < 0.05). Pigs fed HM1 diet had higher (p < 0.05) WBC concentration than those fed CON and HM2 diets at week 6. Pigs fed HM-supplemented diet had higher (p < 0.05) IgG and IgA concentrations than those fed CON diet at week 12. Pigs fed HM diet noted better (p < 0.05) meat colour and redness value than pigs fed CON diet. Pig fed HM2 reduced (p < 0.05) the lightness value compared with CON group. In conclusion, dietary HM supplementation exerted beneficial effects on growth performance, nutrient digestibility, intestinal microbial balance (increased Lactobacillus counts and decreased E. coli counts), immune response and meat quality.

This study aimed to investigate the effects of polysaccharide from Angelica and Astragalus (AAP) on carbon tetrachloride (CCl4) induced liver damage in mice. A total of 120 Kunming mice were randomly distributed among 6 groups comprising (i) the normal control mice, (ii) the CCl4 treatment group, (iii) the bifendate treatment group, (iv) the AAP treatment group, (v) the Angelica sinensis polysaccharide (ASP) treatment group, and (vi) the Astragalus membranaceus polysaccharide (AMP) treatment group. AAP, ASP and AMP were administered to mice treated with CCl4. The activities of alanine transaminase (ALT) and aspartate transaminase (AST) in the serum, and superoxide dismutase (SOD) and malondialdehyde (MDA) in the liver tissues were quantified, as well as the liver index. Hepatic histological changes were observed by staining liver sections with hematoxylin and eosin. Our results show that bifendate, AAP, ASP, and AMP significantly decreased the activities of MDA, AST, and ALT, and enhanced the activity of SOD in CCl4-treated mice. Bifendate, AAP, ASP, and AMP consistently ameliorated the liver injuries induced with CCl4. Notably, the hepatoprotective effect of AAP was stronger than that of bifendate, ASP, or AMP. In addition, AAP alleviated liver inflammation and decreased the liver indexes of mice induced with CCl4. These effects were at least partly due to the antioxidant properties of AAP in scavenging free radicals to ameliorate oxidative stress and to inhibit lipid peroxidation.

UNASSIGNED

Medicinal plants have a long history of use in China to treat diabetic symptoms. Ancient Chinese medical manuscripts and ethnobotanical surveys document plant remedies that continue to be actively used in China for the treatment of diabetic symptoms. Based on a systematic ancient Chinese medical manuscripts review in combination with ethnobotanical survey, 16 medicinal plants for the traditional treatment of diabetic symptoms were identified for the evaluation of anti-insulin resistance bioactivity. The biological activity of 16 medicinal plants was tested on dexamethasone (DXMS)-induced insulin resistant HepG2 cells. The result shows that 11 of the 16 medicinal plants enhanced glucose uptake of DXMS-induced insulin resistant HepG2 cells, thereby demonstrating their ability to increase insulin sensitivity, other five medicinal plants including Astragalus membranaceus were found ineffective. The study shows that ancient Chinese medical manuscripts and ethnobotanical surveys on plants for the prevention and treatment of diabetic symptoms provide a promising knowledge base for drug discovery to mitigate the global diabetes epidemic.

BACKGROUND

The response of conventional chemotherapy for osteosarcoma treatment is usually poor, and chemotherapy-related severe side effects and drug resistance remain a problem. Abundant evidence has shown that Astragaloside IV, extracted from Astragalus membranaceus Bunge, strongly inhibits the growth of many carcinomas. We aimed to investigate the chemosensitive effects of Astragaloside IV in osteosarcoma in vitro and in vivo.

METHODS

Human osteosarcoma cell lines MG-63 and 143B, and BALB/c nu/nu mice xenograft were used. MTT, Clonogenic assay, Annexin V/PI assay and Western bloting analysis were carried out.

RESULTS

Our present study found that Astragaloside IV was a critical chemosensitizing agent for osteosarcoma treatment. Astragaloside IV suppressed cell proliferation and enhanced chemosensitivity in osteosarcoma cell lines and xenograft. Caspase-dependent Fas/FasL signaling was involved in cisplatin-induced apoptosis which was enhanced by Astragaloside IV.

CONCLUSIONS

It indicated that Astragaloside IV might be a promising therapeutic agent for osteosarcoma treatment.

Three new flavonoid glycosides, (3R)-(+)-isomucronulatol-2'-O-β-D-glucopyranoside (1), (3R)-(-)-isomucronulatol-7-O-β-D-apiofuranosyl(1→2)-β-D-glucopyranoside (2), and (2S)-(-)-7,8-dihydroxylflavanone-4'-O-β-D-apiofuranosyl(1→2)-β-D-glucopyranoside (3), along with eight flavanones (4, 8, 10, 12, 15, 16, 21, and 24), four isoflavones (5, 11, 13, and 23), four chalcones (6, 14, 17, and 18), two isoflavans (19-20), one flavone (7), one flavonol (9), and one dihydrochalcone (22) were isolated from Baoyuan Decoction (BYD), a traditional Chinese medicine formula. The structures of the new compounds were established by detailed analysis of NMR and HRESIMS spectroscopic data, and their absolute configurations were determined by electronic circular dichroism (ECD) data. The inhibitory effects of the isolates were evaluated on nitric oxide production in lipopolysaccharide activated RAW 264.7 macrophage cells. Compounds 6, 9, and 10 showed the significant inhibitory activities, with IC50 values of 1.4, 13.8, and 9.3 μM, respectively, comparable to or even better than the positive control, quercetin (IC50, 16.5 μM). The assignment of these isolated flavonoids was achieved using UPLC-Q-trap-MS, and the results suggested that they were originated from Astragalus membranaceus and Glycyrrhiza uralensis.

With the increasing cultivation of medicinal legumes in agricultural fields, the rhizobia associated with these plants are facing new stresses, mainly from fertilization and irrigation. In this study, investigations on the nodulation of three cultivated medicinal legumes, Astragalus mongholicus, Astragalus membranaceus and Hedysarum polybotrys were performed. Bacterial isolates from root nodules of these legumes were subjected to genetic diversity and multilocus sequence analyses. In addition, the distribution of nodule bacteria related to soil factors and host plants was studied. A total 367 bacterial isolates were obtained and 13 genospecies were identified. The predominant microsymbionts were identified as Mesorhizobium septentrionale, Mesorhizobium temperatum, Mesorhizobium tianshanense, Mesorhizobium ciceri and Mesorhizobium muleiense. M. septentrionale was found in most root nodules especially from legumes grown in the barren soils (with low available nitrogen and low organic carbon contents), while M. temperatum was predominant in nodules where the plants were grown in the nitrogen-rich fields. A. mongholicus tended to be associated with M. septentrionale, M. temperatum and M. ciceri in different soils, while A. membranaceus and H. polybotrys tended to be associated with M. tianshanense and M. septentrionale, respectively. This study showed that soil fertility may be the main determinant for the distribution of rhizobia associated with these cultured legume plants.

BACKGROUND

Biochanin A and formononetin are O-methylated isoflavones that are isolated from the root of Astragalus membranaceus, and have antitumorigenic effects. Our previous studies found that formononetin triggered growth-inhibitory and apoptotic activities in MCF-7 breast cancer cells. We performed in vivo and in vitro studies to further investigate the potential effect of biochanin A in promoting cell proliferation in estrogen receptor (ER)-positive cells, and to elucidate underlying mechanisms.

METHODS

ERα-positive breast cancer cells (T47D, MCF-7) were treated with biochanin A. The MTT assay and flow cytometry were used to assess cell proliferation and apoptosis. mRNA levels of ERα, Bcl-2, and miR-375 were quantified using real-time polymerase chain reaction. Compared with the control, low biochanin A concentrations (2-6 μM) stimulated ERα-positive cell proliferation (T47D, MCF-7). The more sensitive T47D cells were used to study the relevant signaling pathway.

RESULTS

After treatment with biochanin A, ERα, miR-375, and Bcl-2 expression was significantly upregulated. Additionally, in the in vivo studies, uterine weight in ovariectomized mice treated with biochanin A increased significantly.

CONCLUSIONS

This study demonstrated that biochanin A promoted ERα-positive cell proliferation through miR-375 activation and this mechanism is possibly involving in a miR-375 and ERα feedback loop.

BACKGROUND

Astragalus membranaceus has been clinically used in cerebral ischemia treatment in China and its main component, Astragaloside IV (Ast IV) shows anti-hypoxia activity, but the underlying mechanism has not been clearly clarified. This study was aimed to investigate the effects of Ast IV on hypoxia-induced injury in PC-12 cells as well as the underlying mechanism.

METHODS

Relative miR-124 expression was detected by qRT-PCR. Hic-5 expression was analyzed by qRT-PCR and Western blot. To alter miR-124 and Hic-5 expressions, cells were respectively transfected with miR-124 mimic and pEX-Hic-5. Cell proliferation and apoptosis were measured by BrdU assay and Annexin V-fluorescein isothiocynate (FITC)/propidium iodide (PI) double staining method, respectively. Besides, apoptotic proteins and cell proliferation-associated factors were analyzed by Western blot.

RESULTS

Ast IV alleviated hypoxia-induced injury in PC-12 cells by decreasing apoptosis (P < 0.01). Ast IV inhibited up-regulation of miR-124 induced by hypoxia (P < 0.01). miR-124 mimic impaired the anti-apoptotic effect of Ast IV on PC-12 cells (P < 0.01). Hic-5 expression was significantly down-regulated in miR-124 overexpressed cells (P < 0.001) and Hic-5 overexpression activated Sp1/Survivin signaling pathway (P < 0.001).

CONCLUSIONS

Ast IV could ameliorate hypoxia-induced injury in PC-12 cells by decreasing miR-124 expression and then up-regulating Hic-5 expression.