Beating Myocardial cell cultures of neonatal rats were prepared in vitro and infected with coxsackie B-2 virus. The cells were evaluated in the post-infected period for changes in beating percentage and cytopathic effect (CPE), alterations in the electrical activities by standard microelectrode techniques, and the protective effect of Astragalus membranaceus (AM) on coxsackie B-2 virus-infected neonatal rat myocardial cell cultures was observed. The beating percentage began to decrease in the infected group at 24 hr and only 27.9 +/- 18.6% was beating at 96 hr after virus challenge, premature beats, tachycardia and fibrillation occurred commonly during the experiment. Meanwhile the CPE appeared rapidly from 1+-3+ at the same interval. Resting potential, action potential amplitude, duration and rate of uptake were shown a significant decrease through 24-96 hr (P less than 0.01). In contrast, the beating and electrical activities were nearly normal and less CPE was shown in myocardial cells treated with AM 1 hr after virus challenge through 24-96 hr (P less than 0.05). The results suggest that AM may be valuable in prophylaxis and treatment of acute coxsackie B-2 virus caused myocarditis.

This study was designed to assess the potential effects of Astragalus membranaceus (AM) on the growth performance and antioxidative stress response in bluegill sunfish (Lepomis macrochirus) exposed to challenging cold water temperature conditions. In this regard, fish with an average weight of 43 ± 1 g were divided into four groups and fed daily with an AM-free diet (control), and 1.5, 3, and 4.5 % (w/w) AM-incorporated diets for an 8-week period. Oxidative stress response, biochemical, and growth parameters were measured, and subgroups of fish were exposed to the outside challenging cold pond water temperature (1.6-9.9 °C) with an average of 7.0 ± 0.1 °C beyond the optimal temperature. The results showed that incorporating AM in the diet significantly improved growth performance parameters (body mass gain, specific growth rate, length, condition factor, and feed conversion ratio) and biochemicals (aspartate aminotransferase and alanine transaminase activities, and glucose and cortisol concentrations). In addition, markedly up-regulated superoxide dismutase, glutathione peroxidase, and catalase activities were observed in AM-treated fish groups over the control. Conclusively, feeding AM diets significantly increased (P < 0.05) growth performance and antioxidative stress profiles throughout the entire experiment, and this increase was much more pronounced at 8 weeks after the water temperature began to rise, which can be related to the nature of Bluegill fish as it is known to be a warm water fish. These findings are considered to be of great importance for sustainable aquaculture.

Objective To observe the effect of Astragalus membranaceus (AM) on expression levels of helper T cell 17 (Th17) and its related factors in patients with allergic rhinitis (AR). Methods Peripheral blood mononuclear cells (PBMCs) were sampled from 31 AR patients (recruited in the AR group) and 22 healthy subjects (recruited in the control group). PBMCs were isolated and in-vitro inter- vened by high and low dose AM injection (2 000 and 500 μg/mL) respectively for 24 h. mRNA expression levels of related orphan receptor C (RORC) were detected by real time quantitative PCR (RT-qPCR). Ex- pression levels of IL-17A and IL-22 in the supernatant were measured by ELISA. Expression levels of Th17 cells were measured by flow cytometry. Results Expression levels of Th17, RORC mRNA, IL- 17A, and IL-22 were higher in the AR group than in the control group (P <0. 01). mRNA expression levels of RORC, Th17 and its cytokines were not changed statistically in the AR group and the control group after PBMCs were intervened by low dose AM (P>0. 05). After intervened by high dose AM,mRNA expres- sion levels of RORC decreased statistically in the AR group and the control group (P <0. 05 for the AR group, P <0. 01 for the control group). Meanwhile,the expression levels of Th-17 and its cytokines de- creased in the AR group and the control group with statistical difference (P <0. 01). Conclusions Ex- cessive activation of Th17 is one of key factors-for AR. AM could further inhibit inflammation of AR and control the inflammation state of AR possibly through inhibiting the differentiation of Th17 cells and promo- ting the release of its cytokines.

A novel biotechnology approach by combining deacetylation biocatalysis with elicitation of immobilized Penicillium canescens (IPC) in Astragalus membranaceus hairy root cultures (AMHRCs) was proposed for the elevated production of astragaloside IV (AG IV). The highest AG IV accumulation was achieved in 36-day-old AMHRCs co-cultured with IPC for 60 h, which resulted in the enhanced production of AG IV by 14.59-fold in comparison with that in control (0.193 ± 0.007 mg/g DW). Meanwhile, AG IV precursors were almost transformed to AG IV by IPC deacetylation. Moreover, expression of genes involved in AG IV biosynthetic pathway was significantly up-regulated in response to IPC elicitation. Also, FTIR and SEM showed that cell wall lignification was enhanced following IPC treatment and root surface was likely to be IPC deacetylation site. Overall, dual roles of IPC (biocatalyst and elicitor) offered an effective and sustainable way for the mass production of AG IV in AMHRCs.

A cocultivation system of Astragalus membranaceus hairy root cultures (AMHRCs) and immobilized food-grade fungi was established for the enhanced production of calycosin (CA) and formononetin (FO). The highest accumulations of CA (730.88 ± 63.72 μg/g DW) and FO (1119.42 ± 95.85 μg/g DW) were achieved in 34 day-old AMHRCs cocultured with immobilized A. niger (IAN) for 54 h, which were 7.72- and 18.78-fold higher than CA and FO in nontreated control, respectively. IAN deglycosylation could promote the formation of CA and FO by conversion of their glycoside precursors. IAN elicitation could intensify the generation of endogenous signal molecules involved in plant defense response, which contributed to the significantly up-regulated expression of genes in CA and FO biosynthetic pathway. Overall, the coupled culture of IAN and AMHRCs offered a promising and effective in vitro approach to enhance the production of two health-promoting isoflavone aglycones for possible nutraceutical and pharmaceutical uses.

The present study was performed to investigate the anti-septic effects of Qi-Shao-Shuang-Gan (QSSG), a combination of Astragalus membranaceus saponins (SAM) and Paeonia lactiflora glycosides (GPL), in septic mice induced by cecal ligation and puncture. QSSG was shown to elevate the survival rate of mice, decrease infiltration of polymorphonuclear leukocytes into livers and lungs, lower serum levels of myeloperoxidase, nitric oxide, and lactate dehydrogenase, and decrease mRNA expressions of inducible nitric oxide synthase and interleukin-1β in livers. It also restored the impaired expressions of protein C (PC) mRNA in mouse livers and expressions of thrombomodulin and endothelial PC receptor mRNA in endothelial cells. Neither SAM nor GPL alone could significantly increase the survival rate of septic mice. The findings indicate that QSSG exerts protective action against polymicrobial sepsis by inhibiting systemic inflammatory response and upregulating PC pathway, and there are synergistic effects between SAM and GPL.

OBJECTIVE

To study Astragalus membranaceus absorption characteristic of nitrogen, phosphorus and potassium at different growth stages.

METHODS

Through the field experiment and the sampling analysis, the absorbing capacity of nitrogen, phosphorus and potassium as well as the growth of plant at different growth stages in A. membranaceus were analyzed.

RESULTS

The dry matter accumulation reached 88.22% of the total accumulation in 100-163 days after seedling emergence, The content of N, P, K in the stem was higher than that in the root system. Nitrogen, phosphorus and potassium absorbing capacity in the stem, the leaf and the root systems was N>> K>> P. In the whole growth period, nitrogen accumulation reached the highest, and followed with the accumulation of potassium, and the accumulation of phosphorus was the lowest. In the last phase of the exuberant growth period, the accumulation intensity of nitrogen, phosphorus and potassium reduced. During the harvesting time, the accumulation intensity of nitrogen and potassium increased, and the accumulation intensity of phosphorus remained stable.

CONCLUSIONS

The dry matter accumulation reached the maximum in 100-163 days after seedling emergence. The absorption strength of nitrogen, phosphorus and potassium in aerial part reached the maximum in 100-132 days after seeding emergence, the maximum absorption comes earlier than that of phosphorus and potassium, at that time the needs of nutrients reach the highest. For producing of 100 kg A. membranaceus 2.32 kg N, 0.323 kg P2O5, 1.62 kg K2O are needed to be absorbed from soil and fertilizer.

Streptococcus alactolyticus strain FGM, isolated from chicken cecum, was used to increase the extract yield of polysaccharides during Astragalus membranaceus fermentation. It was previously demonstrated that polysaccharides from fermented A. membranaceus by S. alactolyticus had some similar properties to those from A. membranaceus in terms of its ability to help heal hepatic fibrosis in rats and modulate immunopotentiation of broiler chicken. However, methods to increase the yield of the polysaccharides during fermentation of A. membranaceus are not well understood. In this paper, we investigated the involvement of uridine diphosphate (UDP)-glucose 4-epimerase (galE) and glucan-1,6-α-glucosidase (dexB) during A. membranaceus fermentation through real-time reverse transcription quantitative PCR. The galE and dexB genes of S. alactolyticus were cloned by homology-based cloning and the genome walking method for the first time, and the 3D structure of dexB was analyzed by Swiss-PdbViewer 4.0.1 software. The expression of both the galE and dexB genes in A. membranaceus fermentation was studied using the determined ideal reference gene ldh for transcript normalization. The results showed that these two genes were both highly induced and peaked after 12 h of fermentation. The expression level of galE was stepwise increased from 48 to 72 h, while dexB transcripts were markedly increased at 60 h and decreased by 72 h. These data suggested that dexB and galE of S. alactolyticus strain FGM were involved in the regulation of A. membranaceus fermentation and they might play some roles in the increase of polysaccharides.

OBJECTIVE

Astragalus membranaceus, one of the most common Chinese herbs, is widely used to prevent and treat a variety of diseases. Very few adverse reactions, caused by A. membranaceus, have been reported in the literature. The purpose of this article was to report a case of marked increase in carbohydrate antigen 19-9 (CA19-9) and the formation of liver and kidney cysts following oral administration of A. membranaceus.

METHODS

A 38-year-old woman was found to have a high serum CA19-9 level (156 U/mL) at her routine annual examination. On follow-up, several small cysts were found in her left kidney and liver by CT scan. Her medical history showed that she had taken Astragalus tea every day for 1 month. One month after she stopped taking it, the CA19-9 level decreased to 40·19 U/mL. Ten months later, PET-CT showed that there were no liver and kidney cysts. However, she took Astragalus powder again in the second year and 1 month later her CA19-9 level increased again to more than 1000 U/mL. Several small cysts were again seen in her left kidney and liver by enhanced CT. Her CA19-9 level gradually became normal after she stopped taking the Astragalus powder.

CONCLUSIONS

This case strongly suggests that oral administration of A. membranaceus may lead to increase in CA19-9 and the formation of liver and kidney cysts.

The aim of the study was to analyze histological and morphometric changes in the brain of rats subjected to immobilization stress and to assess the neuroprotective effect of the extract of Astragalus membranaceus (AE). The work was carried out on Wistar rats that were divided into 3 groups (6 animals in each): group 1 — intact rats; group 2 — control animals subjected to immobilization stress for 18 h; group 3 — rats treated orally with AE (50 mg/kg) for 7 days and then subjected to immobilization stress. The level of stress-induced brain damage was assessed by a morphometric method, by measuring the proportions of morphologically intact neurons, hyperchromatic shrunken neurons and neurons with the signs of degeneration in layers III–V of the cerebral cortex and CA1 region of the hippocampus in the standard area of the section. Immobilization stress in rats led to a clearly pronounced neuronal damage in the cerebral cortex and hippocampus. AE was found to limit the stress-induced neuronal damage in the cerebral cortex and the hippocampus: the proportion of hyperchromatic shrunken neurons was reduced 3 and 4.6 times, respectively, compared to control.

OBJECTIVE

To study the effects of fertilizers with the different proportional of nitrogen, phosphorus and potassium on growth and active ingredient of Astragalus membranaceus.

METHODS

Field experiment was conducted based on the D-saturation optimal design with three factors of nitrogen, phosphorus and potassium. The effects on growth and active ingredient of A. membranaceus were analyzed.

RESULTS

Fertilization promoted the seedling growth and provided abundant supply of nutrition for growth of root, yield and accumulation of active ingredient at the later growth stage, and increased the accumulation of dry matter of stem-leaf and root system. The effect of nitrogen, phosphorus and potassium application on the total dry matter accumulation of A. membranaceus was as following: nitrogen>> potassium>> phosphorus; the effect on the stem-leaf dry matter accumulation was as following: nitrogen>> phosphorus>> potassium; the effect on the root dry matter accumulation was as following: nitrogen>> potassium>> phosphorus. Nitrogen, phosphorus and potassium fertilizer increased the root yield of A. membranaceus. Obviously, the effect on the root yield was as following: nitrogen>> potassium>> phosphorus. The application of different proportional with nitrogen, phosphorus and potassium increased the content of polysaccharide and astragaloside, but had no distinct effect on the content of total flavonoids. The effect on the content of polysaccharide was as following: potassium>> phosphorus>> nitrogen, but the effect on the conten,t of astragaloside was as following: nitrogen>> potassium>> phosphorus.

CONCLUSIONS

Nitrogen and potassium fertilizer application had more important effect on growth, yield and the contents of polysaccharide and astragaloside in A. membranaceus. During medicinal plants cultivation process, it should pay attention to the application of nitrogen fertilizer and potassium fertilizer and make balance application of nitrogen, phosphorus and potassium fertilizer.

The cerebral ischemia rabbit model was made by using the occlusion of four vessels. The results showed that TXB2 and cAMP contents in brain tissues and the latter in plasma markedly increased (P < 0.05, P < 0.01), the 6-keto-PGF1 alpha in brain tissues significantly lowered (P < 0.05) in ischemia formed 30 minutes and 45 minutes after reperfusion. After intravenous injection of Astragalus membranaceus (AM) extracts (3.3 g/kg), Huoxuefang (HXFO and Yiqi Houxue Fang (YQHXF) consisted of AM and HXF before ischemia, the marked increase of TXB2 contents after reperfusion was inhibited (P < 0.05) and the 6-keto-PGF1 alpha in brain tissues after reperfusion were increased (P < 0.01) in HXF and YQHXF group, which change the AM extracts didn't have (P < 0.05). HXF could markedly inhibit the increase of cAMP in brain tissues after reperfusion P < 0.05), while the AM extracts and YQHXF couldn't (P>> 0.05). All above-mentioned suggested that the above-mentioned suggested that the balance disorder of TXA2/PGI2 in brain tissues might participate in the occurrence of cerebral reperfusion injury and YQHXF might act against this injury by means of improving the balance of TXA2/PGI2 in brain tissues, which was mainly released by HX drugs of it.

A study on the anti-herpes simplex virus activity of the suppository or ointment form of Astragalus membranaceus(AM) combined with recombinant human interferon alpha 2b(IFN) was carried out in human diploid cell culture. AM is a Chinese herb medicine and have been used in China as a tonic for thousands of years and the IFN was produced from E. coli with 95% purity. Obtained results indicated that the placebo suppository and ointment(without AM and IFN) seemed not to decrease markedly the anti-viral activity of IFN in WISH/VSV assay system. The anti-herpes simplex virus activities of suppository and ointment forms of AM and IFN were shown to be significantly higher than that of IFN alone. It is well known that chronic cervicitis is closely related to papillomavirus, cytomegalovirus as well as herpes virus infections. The AM-IFN suppository is suggested to be used in the treatment of cervicitis and the ointment in the treatment of skin herpes.

A chalcone reductase (CHR) gene was isolated from Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao (A. mongholicus). The full-length cDNA of A. mongholicus CHR, designated as Amchr (GenBank accession No. HM357239), was 1196 bp long. It had a 957 bp open reading frame encoding a 318-amino acid protein of 35 kDa, a 67 bp 5' non-coding region and a 172 bp 3'-untranslated region. The putative AmCHR protein showed striking similarity to CHR from other leguminous species. Two-dimensional structure modeling showed that AmCHR consisted of 45.28% α-helix, 10.38% extended strand and 44.34% random coil. Prediction showed that three-dimensional AmCHR was a global protein containing an aldo-ket-red domain, with a putative Asp-Tyr-Lys-His catalytic tetrad in the center. The AmCHR gene was 1251 bp long, consisting of three exons and two introns. Intron I was 125 bp and intron II was 169 bp long. Southern blot analysis indicated that Amchr belonged to a small multigene family. Under natural conditions, Amchr was expressed differentially in the root, stem and leaf tissues of A. mongholicus, with a preferential expression in the root. The recombinant AmCHR protein was successfully expressed in Escherichia coli strain BL21 with pET42a vector. The result showed that the expressed AmCHR protein had molecular weight of about 35 kDa, which matched the size of the predicted protein by bioinformatic analysis. This study opened avenues towards understanding of the function of AmCHR protein and the role of the Amchr gene in the calycosin-7-O-β-D: -glucoside branch pathway in A. mongholicus.

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

Astragalus membranaceus is a staple of Traditional Chinese Medicine being one of the oldest medicinal herbs listed in the material medica of Chinese herbal medicine. Chinese herbalists have used Astragalus to help the human body fight a variety of diseases. Modern herbalists utilize Astragalus primarily as an immunostimulant to prevent common infection and aid in the recovery following infection. Historically, the biological activities associated with Astragalus have been accounted for, at least in part, to several constituents present in the botanical including saponins and polysaccharides. We propose that in addition to these constituents, compounds from endophytic (or epiphytic) bacteria present in (or on) the roots of Astragalus may have an important biological role. Lipopolysaccharides and lipoproteins are major components of Gram-negative bacteria and highly potent activators of the innate immune response. Our data supports a direct correlation between the level of immune gene induction and the level of lipopolysaccharides/lipoproteins present in the Astragalus extract. We demonstrate that extracts from Astragalus specifically activate Toll-like and NOD-like receptors involved in the recognition and response to bacterial constituents and that removal of the lipopolysaccharide/lipoprotein from the Astragalus extract reduced the level of this response. The results support that many immune enhancing botanicals have established a symbiotic relationship with Gram-negative bacteria and that the immune enhancing effect of these botanical extracts on the body may not only be due to endogenous plant compounds, but endophytic (or epiphytic) bacterial components as well.

Keywords: Astragalus membranceus; Rhizobium; botanical; endophytic bacteria; immune; lipopolysaccharide; lipoprotein.

We investigated if fermentation with probiotic cultures could improve the production of health-promoting biological compounds in Astragalus membranaceus. We tested the probiotics Enterococcus faecium, Lactobacillus plantarum and Enterococcus faecium + Lactobacillus plantarum and applied PacBio single molecule, real-time sequencing technology (SMRT) to evaluate the quality of Astragalus fermentation. We found that the production rates of acetic acid, methylacetic acid, aethyl acetic acid and lactic acid using E. faecium + L. plantarum were 1866.24 mg/kg on day 15, 203.80 mg/kg on day 30, 996.04 mg/kg on day 15, and 3081.99 mg/kg on day 20, respectively. Other production rates were: polysaccharides, 9.43%, 8.51%, and 7.59% on day 10; saponins, 19.6912 mg/g, 21.6630 mg/g and 20.2084 mg/g on day 15; and flavonoids, 1.9032 mg/g, 2.0835 mg/g, and 1.7086 mg/g on day 20 using E. faecium, L. plantarum and E. faecium + L. plantarum, respectively. SMRT was used to analyze microbial composition, and we found that E. faecium and L. plantarum were the most prevalent species after fermentation for 3 days. E. faecium + L. plantarum gave more positive effects than single strains in the Astragalus solid state fermentation process. Our data demonstrated that the SMRT sequencing platform is applicable to quality assessment of Astragalus fermentation.

A study was carried out to examine the effects of forced running exercise in the growing stage in male ICR mice and of Astragalus membranaceus (As) on their immune functions. The mice were divided at 4 weeks of age into 4 groups. The first group of mice received forced running exercise (E-group), the second group was given As (As-group), the third group received the forced running exercise and was given As (E+As-group) and the fourth group was a control receiving no treatment. The exercise received was forced running at 15 m/min on a flat floor without any slope for 60min a day. The mice of groups E and E+As were exercised 5 times a week for 12 weeks. The mice of groups As and E+As were given As p. o. at 200 mg/kg per day (5 days/week) for 12 weeks. The results obtained were as follows: 1. After 12 weeks of forced running exercise, the weight of the anterior tibialis muscle and succinate dehydrogenase activity in the anterior tibialis muscle increased significantly in groups E and E+As compared with the control group. Thymus weight showed a tendency to increase in groups E and E+As as compared with the control group. 2. The potentiation of the phagocytic function of the reticuloendothelial system examined by the carbon clearance method was seen in groups E, As and E+As. 3. Superoxide anion production of peritoneal macrophages significantly increased in groups As and E+As, but not in group E. 4. The acid phosphatase activity of peritoneal macrophages in groups E, As and E+As significantly increased compared with the control group. 5. Interleukin 1 production by macrophages remained in all groups. 6. The proliferation of splenocytes induced by Con A in groups E, As and E+As significantly increased compared with the control group. These results suggested that forced running exercise in the growing stage in mice and the administration of As enhanced immune functions and that they might also intensify the functioning of the host defense system.

BACKGROUND

Astragalus membranaceus is used to manage the deficiency of vital energy in traditional Chinese medicine and confirmed to have many biological functions. Mesenchymal stem cells (MSCs) possess immunosuppressive effects, and are widely used for regenerative medicine and immune disorders.

OBJECTIVE

This study investigated the effects of Astragalus polysaccharides (APS) on umbilical cord-derived MSCs (UCMSCs), including morphology, surface marker expression, proliferation, differentiation, and in-vitro and in-vivo immunosuppressive capacities.

METHODS

MSCs isolated from umbilical cords were used. PG2 injection, a botanically derived drug containing a mixture of APS, was added into the culture medium to prepare PG2-treated UCMSCs. The morphology, surface marker expression, proliferation, and differentiation of UCMSCs were determined. The in-vitro immunosuppressive effects of UCMSCs were examined by peripheral blood mononuclear cell (PBMC) proliferation assay. The in-vivo effects were evaluated by circulatory inflammation-associated cytokine levels in mice with septic peritonitis induced by cecal ligation and puncture (CLP) operation.

RESULTS

Compared with control UCMSCs, UCMSCs had higher population doublings when exposed to PG2-containing medium (P = 0.003). The reduction rates of PBMC proliferation after phytohemagglutinin stimulation increased significantly when UCMSCs were treated with PG2 (P = 0.004). The serum levels of inflammation-associated cytokines, including TNF-α, IL-6, MCP-1, IFN-γ, and IL-1β, were significantly lower at 6h after CLP in the mice receiving PG2-treated UCMSCs.

CONCLUSIONS

Our results demonstrated that PG2 can enhance UCMSC proliferation and their in-vitro and in-vivo immunosuppressive effects. Consequently, UCMSCs can be obtained in earlier passages to avoid senescence, and sufficient cells can be acquired faster for clinical use. With stronger immunosuppressive effects, UCMSCs may treat immune disorders more effectively. Further studies are warranted.

Ethnopharmacological relevance: In traditional Chinese medicine (TCM) theory, "Qi" is classified as energetic essence supporting the life activities in human. "Blood" is categorized as nourishing essence and circulating in the body. "Blood" and "Qi" have an intimate relationship. Astragali Radix (AR; root of Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao) has a broad spectrum of application for "Qi-Blood" enrichment. Astragaloside IV, a major saponin in AR, has therapeutic functions in erythropoietic, cardiovascular and immune systems. However, the efficacy of astragaloside IV in erythrophagocytosis has not been elucidated.

Aim of the study: The possible functions of astragaloside IV in heme iron recycling during erythrophagocytosis in cultured macrophage were elucidated.

Methods: The translational and transcriptional expressions of heme recycling enzymes were determined after incubating of astragaloside IV for 24 hours in cultured macrophage.

Results: In astragaloside IV-treated macrophage, the expressions, both RNA and protein levels, of regulators of heme recycling, e.g. heme oxygenase-1 (HO-1), ferroportin (FPN), biliverdin reductase A and B (BVRA, BVRB), were markedly induced in dose-dependent manners. In parallel, the transcriptional activity of antioxidant response element, cloned within an expression vector as pARE-Luc and transfected in cultured macrophages, was markedly induced after a challenge with astragaloside IV in a dose-dependent manner. Moreover, the translocation of Nrf2, a transcriptional factor in regulating expression of heme recycling protein, was induced by astragaloside IV, leading to an enrichment at nucleus fraction.

Conclusion: Astragaloside IV shed lights in enhancing the expression of heme recycle proteins via Nrf2/ARE signaling pathway.

Keywords: Heme; Nrf2; astragaloside IV; erythrophagocytosis.

OBJECTIVE

Extracorporeal shock wave lithotripsy (ESWL) is has been shown to reduce renal parenchymal injury subject to application of shock wave lithotripsy in our pervious study. To investigate the protective action of three main components from Astragalus membranaceus, including total saponins of astragalus (TSA), total flavonoids of astragalus (TFA) total polysaccharide of astragalus (TPA) in alleviating shock wave induced kidney damage.

METHODS

Sixty four male rabbits were randomly assigned to a control group or to 3 groups that were premedicated with TSA TFA and TPA respectively prior to application of ESWL. Each group of animals underwent shock wave lithotripsy (18 kV) to the right kidneys and received a total of 1500 shocks. Peripheral blood samples were collected to evaluate the levels of plasma endothelin-1 (ET-1), plasma nitric oxide (NO) and serum malondialdehyde (MDA) before and after shock wave treatment. The concentrations of these markers in the treated kidney tissues were also detected 3 days, 7 days and 14 days after application of ESWL. The changes of histopathology and cells ultrastructure were observed through light microscope and electron microscope. Untreated contralateral kidneys were evaluated as controls.

RESULTS

In control serials the levels of ET-1 and MDA were elevated significantly while the level of NO was significantly decreased after application of shock wave lithotripsy (P < 0.05). The comparison between the controls and premedicated groups demonstrated that all these three components especially TSA and TFA significantly inhibited shock wave induced increasing of ET-1 and MDA (P < 0.05). TSA also significantly suppressed the decrease of NO and made the recovery time earlier compare to the results of controls (P < 0.05). However, TFA and TPA had almost no effects on the change of NO. (P>> 0.05). The results in histopathology showed noticeably damage of glomerular and tubular epithelial cells in the treated kidneys in the controls. The histological alterations in the TPA group were similar to those of the controls. These alterations were significantly milder in the TSA and TFA particular the TSA group.

CONCLUSIONS

TFA and TSA, especially TSA seemed to play the key role in alleviating ESWL induced kidney damage.

OBJECTIVE

To set up a suitable IEC-6 migration model for pharmacological research and observe the effect of complex polysaccharide from Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao to IEC-6 cell migration.

METHODS

The main conditions related to the establishment of the model, including the planting density of the cell, the observation time after scratching, the concentration of the auxiliary material Matrigel, the treatment of the serum-starvation, the concentration of alpha-difluoromethylornithine (DFMO), an inhibitor of the cell migration, were investigated respectively; and the effects of the tested medicines on the model were observed.

RESULTS

4 x 10(5) cell/mL was the suitable planting density of the cell in the 6-well plate; at the 24th hour after scratching was the appropriate time to count the migrating cells; and the proper concentration of Matrigel was 5%; the serum-starvation could evidently reduce the migrating cells, so the culture medium should contain the serum; 2.5 - 5 mmol/L DFMO was proper for inhibition of the cell migration. Complex polysaccharide from Astragalus membranaceus and spermidine both can promote cell migration.

CONCLUSIONS

The established model of IEC-6 cell migration was suitable for intestinal epithelial restitution such as the researches on pathophysiological mechanisms is the effects of the medicines on the cell migration.

Effects of a polysacharide from Chinese herbal plant Astragalus membranaceus (APS) on the expression of Epstein-Barr virus (EBV) immediate early proteins Zta, Rta and EA-D in Raji cells were examined. EBV switch from latent to lytic cycle in Raji cells was induced by 12-O-tetradecanoylphorbol-13-acetate (TPA) and sorbol butyrate (SB) and the effects of APS were examined by immunofluorescence, western blotting and flow cytometry. APS in a non-cytotoxic concentration of 30 μg/ml significantly suppressed the expression of Zta, Rta and EA-D during the EBV lytic cycle. Our observations indicate that APS is potentially useful as an anti-EBV drug.