Poor sperm motility is an important cause of male infertility. In an attempt to identify Chinese medicinal herbs that might improve human sperm motility in vitro, we screened water extracts of 18 herbs with a trans-membrane migration method which measured the percentage of sperm that moved across the 5 micron pores of a Nucleopore membrane from a semen-drug mixture into phosphate buffered saline during 2 hours incubation. Astragalus membranaceus was the only herb that showed a significant stimulatory effect. At 10 mg/ml, it increased the motility of sperm in semen to 146.6 +/- 22.6% of control. It also increased the motility of washed sperm to 138.2 +/- 13.8% of control. Purification of the active component(s) from this herb as well as its application in assisted reproduction technology await further investigation.

The efficacy of Astragalus membranaceus (AM) oral liquor combined with routine therapy and routine therapy alone on T-lymphocyte subsets of peripheral blood in viral myocarditis patients have been studied. The results showed that the T-lymphocyte subsets profile and OKT4/OKT8 ratio of peripheral blood were significantly lower in viral myocarditis patients than that in healthy control (P < 0.05, 0.01). Routine therapy combined with AM could significantly enhance OKT3, OKT4 and OKT4/OKT8 ratio in the above-mentioned patients (P < 0.05, 0.01). The possible pharmacodynamic mechanism of improved cell immunity in viral myocarditis patients by combining routine therapy with AM was discussed.

For the first time three different natural compounds, isolated from hairy roots of Astragalus membranaceus, cultivated in airlift bioreactor were tested for their cytotoxic potential and apoptosis induction in a panel of human tumor cell lines. Root cultures, cultivated in bioreactor gave 18.5 g l(-1) dry wt roots with the highest astragaloside production in vitro up to now - 1.64% (astragaloside I), 1.12% (astragaloside II) and 1.08% (astragaloside III). In this manner the production in airlift bioreactor can be used as means of reliable supply of cycloartane saponins to extend the research to human clinical studies.

OBJECTIVE

To explore the influence of Astragalus membranaceus (AM) on serum cytokines, Th1, including interleukin-2 (IL-2) and gamma-interferon (gamma-IFN), and Th2, including interleukin-4 (IL-4) and interleukin-10 (IL-10), in patients with herpes simplex keratitis (HSK).

METHODS

One hundred and six HSK patients were randomly divided into the AM treated group and the ribavirin treated group. Levels of serum IL-2, IL-4, IL-10 and gamma-IFN of all the patients and 62 healthy person, selected from donors for control group, were determined by sandwich enzyme-linked immunosorbent assay (ELISA) technique.

RESULTS

Levels of serum IL-4 and IL-10 in HSK patients were significantly higher and those of IL-2 and gamma-IFN were significantly lower than those in the healthy control (all P < 0.01). These parameters were significantly improved in the patients of the AM group after treatment, but with no change in patients of the ribavirin group.

CONCLUSIONS

AM can modulate the imbalance state of Th1/Th2 in HSK patients, improve their immune function disturbance, that shows important significance in treating HSK.

Twelve compounds, including six sucrose fatty acid esters (1-6), four galactosyl acylglycerols (7-10), and two sphingolipids (11 and 12), were isolated from the roots of Astragalus membranaceus . Their structures were identified on the basis of spectroscopic analysis. Among the isolated sucrose fatty acid esters, 6'-O-linoleyl sucrose (1) was identified as a new compound, and 6'-O-palmitoyl sucrose (2) and 6-O-palmitoyl sucrose (3) were isolated from nature for the first time. This is the first report on sucrose fatty acid ester components from A. membranaceus. The nuclear factor-κB (NF-κB) inhibitory activity of isolated compounds was measured in HepG2 cells stimulated with TNF-α using a luciferase reporter system. Among them, compounds 1-6 exhibited significant inhibition of NF-κB activation in a dose-dependent manner, with IC50 values ranging from 4.4 to 24.7 μM. Compounds 1-6 also exhibited inhibition of TNF-α-induced expression of iNOS and ICAM-1 mRNA and dose-dependent inhibition of iNOS promoter activity, with IC50 values ranging from 3.3 to 5.0 μM. These data demonstrate the potential of sucrose fatty acid esters from A. membranaceus to prevent and treat inflammatory diseases.

(3R)-(-)-7,2'-Dihydroxy-3',4'-dimethyl isoflavan-7-O-beta-D-glucopyranoside and (6aR, 11aR) 9,10-di-methoxypterocarpan-3-O-beta-D-glucopyranoside were separated from the ethyl acetate extract of the root of Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao by high-speed counter-current chromatography (HSCCC). A two-phase system composed of ethyl acetate-ethanol-acetic acid-water (4:1:0.25:5, v/v) was selected by analytical HSCCC. Preparative HSCCC yielded, from 100 mg of the partially purified extract, 50 mg of isoflavan glycoside and 10 mg of pterocarpan glycoside each at over 95% purity by high-performance liquid chromatography (HPLC) analysis. Their structures were identified by MS, 1H NMR and 13C NMR.

Telomeres are structures at the ends of chromosomes that shorten during cell division and eventually signal an irreversible state of growth arrest known as cellular senescence. To delay this cellular aging, human T cells, which are critical in the immune control over infections and cancer, activate the enzyme telomerase, which binds and extends the telomeres. Several different extracts from the Astragalus membranaceus root have been documented to activate telomerase activity in human T cells. The objective of this research was to compare two extracts from Astragalus membranaceus, TA-65 and HTA, for their effects on both telomerase and proliferative activity of human CD4 and CD8 T cells. Our results demonstrate that, TA-65 increased telomerase activity significantly (1.3 to 3.3-fold relative to controls) in T cell cultures from six donors tested, whereas HTA only increased telomerase levels in two out of six donors. We also demonstrate that TA-65 activates telomerase by a MAPK- specific pathway. Finally, we determine that during a three-day culture period, only the T cells treated with the TA-65 extract showed a statistically significant increase in proliferative activity. Our results underscore the importance of comparing multiple telomerase activators within the same experiment, and of including functional assays in addition to measuring telomerase activity.

Astragalus membranaceus polysaccharide-iron (III) complex (APS-iron) was synthesized and characterized. Based on single factor and response surface optimization experiments of APS-iron (III) complex synthesis, the optimum conditions of APS-iron (III) complex were obtained as follows: the reaction temperature 89.46°C, pH 8.16, reaction time 46.04min and ratio of catalyst to APS 0.75, respectively. The reaction temperature was the most significant factor, followed by pH, reaction time and the ratio of catalyst to APS in the four reaction parameters. The highest iron content (19.32%) of APS-iron (III) complex was obtained at the optimum conditions, which was characterized by fourier transform infrared (FTIR), scanning electron microscopy (SEM), antioxidant activities of the APS-iron (III) complex and iron release of APS-iron (III) complex in vitro assay. The results indicated the APS-iron (III) complex had good bioavailability and antioxidant activities in vitro assays. So, it was potential for APS-iron (III) complex as a candidate for iron supplements.

Astragalus membranaceus (AM), a traditional Chinese medicinal herb, has immunoregulatory properties in many diseases. We investigated the effects and mechanism of Astragalus membranaceus extract (AME) in the macrophage migration and immune response mediator release. The viability of Ana-1 macrophages treated with AME was evaluated by the MTT method. The secretion and mRNA levels of IL-1β and TNF-a were measured by ELISA and RT-PCR, respectively. Macrophage migration was assayed by transwell assay. The activity of heparanase (HPA) was determined by a heparin-degrading enzyme assay. Our results didn't show any toxicity of AME in macrophages. AME increased the activity of HPA, cell migration, mRNA levels and secretion of IL-1β and TNF-a in macrophages. Pretreatment with anti-HPA antibody reduced cell migration, secretion of IL-1β and TNF-a did not change the mRNA levels of IL-1β and TNF-a significantly in AME-treated macrophages. This suggests that AME may increase the release of immune response mediator and cell migration via HPA to activate immune response in macrophages.

An oxidative stress insult is one of the principal causes of Parkinson's disease. Astragaloside IV (AS-IV), a constituent extracted from Astragalus membranaceus, has been demonstrated to exert antioxidant effects. However, the mechanisms responsible for the antioxidant properties and neuroprotective effects of AS-IV remain unclear. In this study, we examined the protective effects of AS-IV against the apoptosis of human neuronal cells (SH-SY5Y cells) induced by hydrogen peroxide (H2O2). The results revealed that AS-IV pre-treatment attenuated the H2O2‑induced loss of SH-SY5Y cells in a dose-dependent manner; AS-IV exerted significant protecitve effects by decreasing the apoptotic ratio and attenuating reactive oxygen species overproduction in H2O2-exposed SH-SY5Y cells. By means of immunofluorescence staining, AS-IV was found to decrease the expression of α-synuclein and to increase the expression of tyrosine hydroxylase (TH) in the cells, which had been increased and decreased, respectively by H2O2. As shown by western blot analysis, the protective effects of AS-IV against SH-SY5Y cell injury induced by H2O2 were also mediated via the downregulation of the ratio of Bax/Bcl-2. We found that the neuroprotective effects of AS-IV were associated with the inhibition of the expression of the α-synuclein via the p38 mitogen-activated protein kinase (MAPK) signalling pathway. On the whole, our results suggest that AS-IV exerts protective effects against neurodegenerative diseases by targeting α-synuclein or TH.

In this work, Astragalus membranaceus hairy root cultures (AMHRCs) were exposed to ultraviolet radiation (UV-A, UV-B, and UV-C) for promoting isoflavonoid accumulation. The optimum enhancement for isoflavonoid production was achieved in 34-day-old AMHRCs elicited by 86.4 kJ/m(2) of UV-B. The resulting isoflavonoid yield was 533.54 ± 13.61 μg/g dry weight (DW), which was 2.29-fold higher relative to control (232.93 ± 3.08 μg/g DW). UV-B up-regulated the transcriptional expressions of all investigated genes involved in isoflavonoid biosynthetic pathway. PAL and C4H were found to be two potential key genes that controlled isoflavonoid biosynthesis. Moreover, a significant increase was noted in antioxidant activity of extracts from UV-B-elicited AMHRCs (IC50 values = 0.85 and 1.08 mg/mL) in comparison with control (1.38 and 1.71 mg/mL). Overall, this study offered a feasible elicitation strategy to enhance isoflavonoid accumulation in AMHRCs and also provided a basis for metabolic engineering of isoflavonoid biosynthesis in the future.

We isolated rat heart mitochondria and induced lipid peroxidation with ADP and FeSO4. Oxygen consumption and MDA formation were measured for quantitating the amount of lipid peroxidation. Using these methods, we screened the water extracts of 14 Chinese medicinal herbs for their effect on lipid peroxidation. It was found that Astragalus membranaceus inhibited 42.1 +/- 3.4% of oxygen consumption and 39.8 +/- 3.2% of MDA production at concentration of 2 mg dried herb/ml mitochondrial suspension. At the same concentration, Polygonum multiflorum inhibited 52.1 +/- 7.3% of oxygen consumption and 50.9 +/- 5.3% of MDA production. Other herbs did not inhibit lipid peroxidation to 50% of control at concentration up to 6 mg dried herb/ml mitochondrial suspension. Purification and identification of the active component(s) in Astragalus membranaceus and Polygonum multiflorum as well as their clinical application await further studies.

OBJECTIVE

To evaluate the preliminary efficacy and safety of the mixture of drug extracts from 5 Chinese medicinal herbs (SH), in the treatment of Human Immunodeficiency Virus (HIV) infection among people living with HIV/AIDS (PLWHA).

METHODS

Open-label study.

METHODS

Sanpatong Hospital, Chiang Mai, Thailand.

METHODS

HIV-1 infected adults with a CD4 cell count of more than 200 cell/mm3 and HIV-1 RNA>> 20,000 copies/ml.

METHODS

Patients received an oral suspension of SH, a combination of 5 Chinese medicinal herbs namely Glycyrrhiza glaba L., Artemisia capillaris Thumb., Morus alba L., Astragalus membranaceus(Fisch.) Bge., Carthamus tinctorius L., 5 g or 30 ml, in 3 divided doses after meals, plus sulfamethozaxole/ trimethoprim, 400/80 mg tablet, once daily after breakfast for 12 weeks. During the treatment and the follow up period, the absolute CD4 cell count and the plasma HIV-1 RNA were monitored. Adverse events were observed.

RESULTS

Of the 28 enrolled patients, the number of positive response patients with reduction of plasma HIV-1 RNA more than 0.5 log during the treatment and follow up period were 4-10 (14.2-35.7%) while the number of negative response patients who had plasma HIV-1 RNA rising at least 0.5 log were 2-4 (0-14.2%). The means viral load at week 0 (baseline), 12 and 20 were 4.94, 4.83 and 4.76 log copies/ml, which were slightly declined Whilst, the mean absolute CD4 cell count of week 0 (baseline), 4, 8, 12, and 20 fluctuated within the baseline, range of 382.1, 404.2, 359.4, 404.1, 360.2 cell/mm3, respectively. All subjects had good compliance without any serious adverse events.

CONCLUSIONS

Under the condition used, SH drug therapy is safe. Satisfactory positive response, by decreased viral load of more than 0.5 log, was found in 14%-35% of HIV-positive patients. However, the immunologic response, an increase of CD4 cell count was not clearly demonstrated. The clinical benefit of SH needs more thorough scientific support before being prescribed as adjunctive therapy for treating PLWHA.

OBJECTIVE

To study the effects of astragalus membranaceus injection on sperm abnormality in Cd-induced rats.

METHODS

Thirty adult male Sprague-Dawley rats were divided into 5 groups: low concentration abstragalus membranaceus group(A1), high concentration astragalus membranaceus group(A2), cyclophosphamide group (CP), cadmium chloride group (Cd) and control group (C). Astragalus membranaceus injections 5 g/(kg.d) or 10 g/(kg.d) were given intraperitoneally to the rats of the A1, A2, CP and Cd groups. After 7 days, cadmium chloride(0.2 mg Cd/kg body weight) was administrated by intraperitoneal injection stimultaneously to the A1, A2 and Cd group rats. At 22 d after cadmium chloride induction, the rats were sacrificed and the testis coefficient, testicular sperm count, daily sperm production, epididymal sperm count and percentage of sperm abnorality were observed, and the testes and epididymides were studied pathologically.

RESULTS

Testis coefficient, testicular sperm count, daily sperm production and epididymal sperm count in the A2 group [(5.68 +/- 1.19), (49.01 +/- 8.78) x 10(6)/g, (10.25 +/- 2.30) x 10(6)/(g.d), (47.51 +/- 22.51) x 10(6)/ml] increased significantly compared with those of the Cd group [(3.11 +/- 0.16), (37.59 +/- 10.63) x 10(6)/g, (5.31 +/- 0.32) x 10(6)/(g.d), (10.89 +/- 2.45) x 10(6)/ml] (P < 0.05 or P < 0.01). The percentage of sperm abnormality in the A2 group [(7.04 +/- 0.12)%] decreased significantly compared with that of the Cd group [(17.81 +/- 1.55)%] (P < 0.01).

CONCLUSIONS

As an antimutagen astragalus membranaceus has practical value in occupational protection against Cd-induced genetic damage.

ACE inhibitors (ACEi) reduce renal tubulointerstitial fibrosis but are not completely effective. Combined extract of Astragalus membranaceus and Angelica sinensis (A&A) is a traditional antifibrotic agent in China. The present investigation aimed to determine whether an ACEi (Enalapril) and A&A together have a better antifibrotic effect in unilateral ureteral obstruction (UUO) than monotherapy with either agent. Male Sprague-Dawley rats (N = 4 per group) had either sham operation or UUO alone, with A&A (combined aqueous and ethanol extract equivalent to 2.1 g dried herbs), with Enalapril (in drinking water at 200 mg/mL) or with both treatments. Kidney and liver were collected for protein extraction or fixed for histologic stains, immunohistochemistry (IHC), microscopy. Enalapril or A&A individually were antifibrotic. Transforming growth factor-beta1, fibroblast activation, collagen deposition, macrophage accumulation and tubular cell apoptosis were all decreased. The combination of the two drugs was significantly more effective than Enalapril alone in reducing tumor necrosis factor-alpha, collagen accumulation, activation of fibroblasts, and tubular cell apoptosis. In conclusion, Enalapril with A&A significantly decreased tubulointerstitial fibrosis to a greater extent than treatment with Enalapril alone. Further studies focusing on the isolation of the active constituents of A&A and the clinical application of the combination of ACEi plus A&A are warranted to determine the value of this treatment in humans.

Tumor-associated macrophages (TAMs) in tumor microenvironment regulate cancer progression and metastases. In breast cancer, macrophage infiltration is correlated with a poor prognosis. While metastatic breast cancer is poor prognostic with a severe mortality, therapeutic options are still limited. In this study, we demonstrate that KSG-002, a new herbal composition of radices Astragalus membranaceus and Angelica gigas, suppresses breast cancer via inhibiting TAM recruitment. KSG-002, an extract of radices Astragalus membranaceus and Angelica gigas at 3 : 1 ratio, respectively, inhibited MDA-MB-231 xenograft tumor growth and pulmonary metastasis in nude mice, while KSG-001, another composition (1 : 1 ratio, w/w), enhanced tumor growth, angiogenesis, and pulmonary metastasis, in vivo. KSG-002 further decreased the infiltrated macrophage numbers in xenograft tumor cohorts. In Raw264.7 cells, KSG-002 but not KSG-001 inhibited cell proliferation and migration and reduced TNF-alpha (TNF α ) production by inhibiting NF- κ B pathway. Furthermore, a combinatorial treatment of KSG-002 with TNF α inhibited a proliferation and migration of both MDA-MB-231 and Raw264.7 cells. Taken together, we conclude that KSG-002 suppresses breast cancer growth and metastasis through targeting NF- κ B-mediated TNF α production in macrophages.

OBJECTIVE

To investigate the effects on myocardial injury and sarcoplasmic reticulum (SR) Ca(2+)-ATPase of viral myocarditis mice treated with Astragaloside (AS) and Astragalus Injection (AI).

METHODS

Viral myocarditis model was created by intraperitoneal inoculation with coxsackievirus B3m (CVB3m) solution and were divided into model, AS, AI and normal control groups. The mortality, myocardial pathological changes, serum cardiac troponin I (cTnI) and the activity of myocardial Sarco/Endoplasmic Ca(2+)-ATPase (SERCA) were observed.

RESULTS

The mortality of model was higher than that of the normal control (P = 0.0042), AS and AI (P < 0.05). The serum level of cTnI of model was significantly higher than that of the normal control (P < 0.001), AS (P < 0.025) and AI (P < 0.05). The myocardial necrosis and inflammatory changes of AS and AI groups were alleviated than that of model (P < 0.01). The activity of myocardial SERCA of model were significantly lower than that of normal control (P < 0.001), AS (P < 0.01) and AI (P < 0.05).

CONCLUSIONS

AS and AI have some protecting effects on myocardial injury of viral myocarditis mice. AS is the effective component of Astragalus membranaceus in treating viral myocarditis. One of the mechanisms of Astragalus membranaceus and AS for viral myocarditis mice depriving of the myocardial injury may be due to improve the activity of myocardial SERCA in the mice.

BACKGROUND

Clinical research encompasses a wide variety of disciplines. Traditional Chinese medicine (TCM) and natural product research have made great contributions to preventing and treating illness. The number and content of original research reports evaluating TCM and natural products have not previously been described. Information in this area will identify areas of relative strength and weakness in terms of knowledge gaps with respect to clinical conditions and natural product remedies.

METHODS

Original research reports (i.e. original articles, brief reports, and research letters) published in the Chinese Medical Journal (CMJ) from January 2000 to December 2009 evaluating TCM and other natural products were reviewed. The United Kingdom Clinical Research Collaboration (UK-CRC) Health Research Classification System was used to analyze the type of health research conducted. Further analysis on the major illnesses addressed and the major herbal components utilized was conducted.

RESULTS

One hundred and seventeen original research reports involving TCM or other natural products were identified, comprising 3.82% of the CMJ output in the period covered by this study. Of the different materia medica described in these reports, 74.4% were derived exclusively from plant material, 10.3% from animals, 3.4% from fungi, 1.7% from minerals, and 10.3% were of mixed (plant/animal/fungal/mineral) composition. Twelve herbs were investigated exclusively or were constituents of 66/87 (75.9%) of the plant-based materia medica investigated. Panax ginseng was the most commonly investigated herb or constituent (14/87, 16.1%), followed by Astragalus membranaceus (9/87, 10.3%), Coptis chinensis/Berberis spp. (7/87, 8.0%) and Rheum spp. (7/87, 8.0%). Four UK-CRC health categories accounted for the majority of TCM and other natural product research (cancer, 20.9%; cardiovascular, 19.2%; oral/gastrointestinal, 9.8%; and inflammatory/immune, 9.0%). The most common research activity was "development of treatments and therapeutic interventions", which was undertaken by 103/117 (88.0%) of the research investigations reported. Human clinical trials involving natural products accounted for only 5.31% of the reported studies.

CONCLUSIONS

This is a relatively early systematic description of published research from a single journal related to TCM and other natural products. The majority of the research reports described preliminary findings and very few controlled clinical trials in human subjects were reported. Further applied studies will be required to demonstrate the clinical effectiveness, utility and cost-effectiveness of TCM and natural products in clinical practice. The UK-CRC health research classification system is a useful tool for evaluating published research output and could be applied to describe the output from other journals, national and provincial funding bodies, charities, and non-governmental organizations involved in supporting health-related research.

BACKGROUND

It has been reported that formononetin (FMN), one of the main ingredients from famous traditional Chinese medicine "Huang-qi" (Astragalus membranaceus [Fisch] Bunge) for Qi-tonifying, exhibits the effects of immunomodulation and tumor growth inhibition via antiangiogenesis. Furthermore, A. membranaceus may alleviate the retinal neovascularization (NV) of diabetic retinopathy. However, the information of FMN on retinal NV is limited so far. In the present study, we investigated the effects of FMN on the hypoxia-induced retinal NV and the possible related mechanisms.

METHODS

The VEGF secretion model of acute retinal pigment epithelial-19 (ARPE-19) cells under chemical hypoxia was established by the exposure of cells to 150 μM CoCl2 and then cells were treated with 3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole (YC-1, a potent HIF-1α inhibitor, 1.0 μg/mL) or different concentrations of FMN (0.2 μg/mL, 1.0 μg/mL, and 5.0 μg/mL). The supernatants of cells were collected 48 hours later to measure the VEGF concentrations, following the manufacturer's instruction. The mRNA expressions of VEGF, HIF-1α, PHD-2, and β-actin were analyzed by quantitative reverse transcription polymerase chain reaction, and the protein expressions of HIF-1α and PHD-2 were determined by Western blot analysis. Furthermore, the rats with retinopathy were treated by intraperitoneal administration of conbercept injection (1.0 mg/kg) or FMN (5.0 mg/kg and 10.0 mg/kg) in an 80% oxygen atmosphere. The retinal avascular areas were assessed through visualization of the retinal vasculature by adenosine diphosphatase staining and hematoxylin and eosin staining.

RESULTS

FMN can indeed inhibit the VEGF secretion of ARPE-19 cells under hypoxia, downregulate the mRNA expression of VEGFA and PHD-2, and decrease the protein expression of VEGF, HIF-1α, and PHD-2 in vitro. Furthermore, FMN can prevent hypoxia-induced retinal NV in vivo.

CONCLUSIONS

FMN can ameliorate retinal NV via the HIF-1α/VEGF signaling pathway, and it may become a potential drug for the prevention and treatment of diabetic retinopathy.

Astragali Radix (AR) is a commonly used herbal drug in traditional chinese medicine and is widely used for the treatment of diabetes, cardiovascular diseases, nephropathy, and neuropathy. The main source of AR in China is the dried root of Astragalus membranaceus var. mongholicus (Bge.) Hsiao, and both cultivated and wild ARs are used clinically. A systematic comparison of cultivated AR (GS-AR) and wild AR (SX-AR) should be performed to ensure the clinical efficacy and safety. In this study, the chemical composition of the two different ARs, which were collected in the Shanxi (wild) and Gansu (cultivated) provinces, were compared by NMR-based metabolic fingerprint coupled with multivariate analysis. The SX-AR- and GS-AR-induced metabolic changes in the endogenous metabolites in mice were also compared. The results showed that SX-AR and GS-AR differed significantly not only in the primary metabolites but also in the secondary metabolites. However, alterations among the endogenous metabolites in the serum, lung, liver, and spleen were relatively small. This study provided a novel and valuable method for the evaluation of the consistency and diversity of herbal drugs, and further studies should be conducted on the difference in polysaccharides as well as the biological effects between the two kinds of AR.

BACKGROUND

Astragalus membranaceus Bunge is one of the oldest and most frequently used crude herbs in traditional Chinese medicine. The total flavonoids of Astragalus (TFA) are the main active components isolated from Astragalus membranaceus Bunge. Our recent study has shown its potential immunomodulatory and anti-inflammatory effects in vivo and in vitro. However, its anti-arthritic effects and mechanisms of action involved have not been elucidated. The aim of this study was to evaluate the protective effects and possible mechanisms of TFA on Freund's complete adjuvant (FCA)-induced arthritis in rats.

METHODS

Wistar rats were intradermally injected FCA into the right hind metatarsal footpads to establish adjuvant-arthritic model. The rats were intragastrically administered daily with TFA at 25, 50 and 100mg/kg for 28days after FCA induction. Body weight, primary paw swelling, arthritis index, thymus and spleen indices were measured. The levels of serum tumor necrosis factor (TNF)-α, interleukin (IL)-1β, prostaglandin (PG)E2, osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) were determined using ELISA. Histopathological changes and scores in joint tissues were examined using hematoxylin and eosin (H&E). The expression of nuclear factor (NF)-κB p65 in synovial tissues was assayed using immunohistochemical method.

RESULTS

TFA significantly increased body weight, attenuated primary paw swelling and arthritis index, decreased thymus and spleen indices of rats induced by FCA. Furthermore, TFA significantly inhibited serum TNF-α, IL-1β, PGE2 and RANKL production, and promoted serum OPG production and OPG/RANKL ratio of rats induced by FCA. Histopathological examination indicated that TFA significantly attenuated inflammatory cell infiltration, synovial hyperplasia, pannus formation, and bone and cartilage damage. Immunohistochemical assay indicated that TFA inhibited NF-κB p65 expression in synovial tissues of rats induced by FCA.

CONCLUSIONS

These results suggest that TFA exerts potential protective effects against FCA-induced arthritis in rats by regulating OPG/RANKL/NF-κB pathway.

BACKGROUND

Astragaloside Ⅳ (ASG-Ⅳ, (Fig. 1) is the most active component of Chinese sp. Astragalus membranaceus Bunge (Fabaceae) that has showed antioxidant, antiapoptotic and antiviral activities among others. It is reported to play an important role in cardiac fibrosis (CF), but the mechanism remains unclear.

OBJECTIVE

To investigate the mechanism of ASG-Ⅳ on inhibiting myocardial fibrosis induced by hypoxia.

METHODS

We studied the relationship between anti-fibrotic effect of ASG-Ⅳ and transient receptor potential cation channel, subfamily M, member 7 (TRPM7) by in vivo and in vitro experiments.

METHODS

In vivo, CF was induced by subcutaneous isoproterenol (ISO) for 10 days. Rat hearts were resected for histological experiment and reverse transcription real-time quantitative poly merase chain reaction (RT-qPCR). In vitro, molecular and cellular biology technologies were used to confirm the anti-fibrosis effect underlying mechanism of ASG-Ⅳ.

RESULTS

Histological findings and the collagen volume fraction showed that ASG-Ⅳ decreased fibrosis in heart tissues. Hypoxia could stimulate the proliferation and differentiation of cardiac fibroblast which indicated that the degree of fibrosis was increased significantly. Anoxic treatment could also obviously up-regulate the expression of TRPM7 protein and current. ASG-Ⅳ groups showed the opposite results. Knock-down TRPM7 experiment further confirmed the role of TRPM7 channel in hypoxia-induced cardiac fibrosis.

CONCLUSIONS

Our results suggest that the inhibition of hypoxia-induced CF in vivo and in vitro by ASG-IV is associated with reduction of the expression of TRPM7. The moderate inhibition of the TRPM7 channel may be a new strategy for treating cardiac fibrosis.

BACKGROUND

The mechanism of mucosal damage induced by ischemia-reperfusion (IR) after hemorrhagic shock is complex; mast cells (MC) degranulation is associated with the mucosal damage. Astragalus membranaceus can protect intestinal mucosa against intestinal oxidative damage after hemorrhagic shock, and some antioxidant agents could prevent MC against degranulation. This study aimed to observe the effects of astragalus membranaceus injection on the activity of intestinal mucosal mast cells (IMMC) after hemorrhage shock-reperfusion in rats.

METHODS

Thirty-two Wistar rats were randomly divided into the normal group, model group, low dosage group, (treated with Astragalus membranacaus injection, 10 g crude medication/kg) and high dosage group (treated with Astragalus membranacaus injection, 20 g crude medication/kg). The rat model of hemorrhagic shock-reperfusion was induced by hemorrhage for 60 minutes followed by 90 minutes of reperfusion. The animals were administrated with 3 ml of the test drug solution before reperfusion. At the end of study, intestinal pathology, ultrastructure of IMMC, and expression of tryptase were assayed. The levels of malondisldehyde (MDA), TNF-alpha, histamine, and superoxide dismutase (SOD) activity in intestine were detected, and the number of IMMC was counted.

RESULTS

The Chiu's score of the rats in the model group was higher than in other groups (P < 0.01). The Chiu's score in the high dosage group was higher than that in the low dosage group (P < 0.05). Hemorrhage-reperfusion induced IMMC degranulation: Astragalus membranaceus injection attenuated this degranulation. Expression of tryptase and the number of IMMC in the model group increased compared with the other groups (P < 0.01) and was significantly reduced by the treatments of Astragalus membranaceus injection at both doses. There was no significant difference between the two treatment groups (P>> 0.05). MDA content and concentration of TNF-alpha in the model group were higher than that in the other three groups (P < 0.05), and the concentration of TNF-alpha in the low dosage group was higher than that in the high dosage group (P < 0.05). SOD activity and the concentration of histamine in the model group were lower than the other three groups (P < 0.05). There was a negative correlation between the Chiu's score and the concentration of histamine and a positive correlation between the Chiu's score and the concentration of TNF-alpha and between the SOD activity and the concentration of histamine in the four groups (P < 0.05).

CONCLUSIONS

Astragalus membranaceus injection may reduce the damage to small intestine mucosa by inhibiting the activated IMMC after hemorrhagic shock.