According to Traditional Chinese Medicine (TCM) theories, TCM with different meridian tropism have different therapeutic effects. In view of the meridian tropism of Astragalus membranaceus (Huangqi), astragaloside IV, one of the effective phytochemicals of Huangqi, was appointed and observed its distribution in rat tissues following a single intravenous (i.v.) dose. A simple and accurate LC-ESI-MS/MS method was developed and validated for astragaloside IV quantification in heart, liver, spleen, lung and kidney using warfarin as an internal standard (IS). Chromatographic separation was performed on a Eclipse plus C18 (4.6mm×100mm, 1.8μm) when the flow rate was set at 0.300mLmin(-1) and ammonium acetate aqueous solution - acetonitrile was used as mobile phase. The intra- and inter-day precisions of the quality control samples were within 15% and accuracies were within 90.0-110%. The recoveries were more than 90.0% at high, medium and low concentrations, respectively. This method was successfully applied for distribution of astragaloside IV after intravenous (i.v.) dose of 4mgkg(-1) astragaloside IV in rats. Astragaloside IV concentration was highest in liver and kidney and remained much higher than that in other tissues over the experiment course. Lung, heart and spleen were also detected to contain astragaloside IV. The results clearly demonstrated that astragaloside IV was one of the material bases of the meridian tropism of Huangqi.

OBJECTIVE

The root of Astragalus membranaceus, regarded as a tonic in traditional Korean medicine, has been prescribed for long periods to treat chronic illness by boosting the immune system. Ultraviolet (UV) irradiation causes damage to skin connective tissue by degrading collagen, which is a major structural component of the extracellular matrix. Such damage is considered to be a cause of the wrinkling observed in premature ageing of the skin. This study has investigated the photo-protective effect of A. membranaceus on UVB radiation-induced activation of nuclear factor kappa-B (NF-κB) activity in human dermal fibroblasts.

METHODS

HS68 fibroblast cells cultured with various concentrations of A. membranaceus were exposed to UVB (40 mJ/cm²). Activation of NF-κB P65 and expression of matrix metalloproteinase-1 (MMP-1) and type 1 procollagen were measured by Western blotting. Translocation of NF-κB P65 and MMP-1 regulation were also examined by immunocytochemistry.

RESULTS

Western blotting and immunocytochemistry results showed that A. membranaceus inhibited UVB-induced translocation of NF-κB P65 and MMP-1 expression. The data suggested that A. membranaceus restored type 1 procollagen synthesis by inhibiting NF-κB P65 activity and MMP-1 expression in UVB-exposed human dermal fibroblasts.

CONCLUSIONS

A. membranaceus is a candidate for use in skin protection from UVB-induced skin inflammation and photoageing.

Inhibition of inflammatory responses, acceleration of basal cell growth and balanced synthesis of the extracellular matrix (ECM) are important in the healing of open cutaneous wounds. To evaluate the wound-healing effects of Astragali Radix (AR) (the root of Astragalus membranaceus [Fisch.]), experimental open wounds were made on the dorsal side of rats under anesthesia. Boiling water extracts of AR, soaked into a hydrophilic foam dressing, were topically applied to the wounds once a day for 11 consecutive days. The healing process was assessed by scoring macroscopic appearance and measuring the area of the open wounds. Molecular aspects of the healing skin area were also investigated via histological observation indicating cell density and linear alignment of the granulation tissue. The AR extracts significantly accelerated cutaneous wound healing by suppressing inflammation and stimulating basal cell growth in the wound area compared to epidermal growth factor as a positive control. Promotion of basal cell proliferation and angiogenesis by the AR extracts was remarkable in the early stages of wound healing, resulting in a significant reduction in the duration of the wound-healing process. We conclude that extracts of AR could be useful in enhancing cutaneous wound healing.

BACKGROUND

Apoptosis plays a critical role in the progression of diabetic cardiomyopathy (DC). Astragalus polysaccharides (APS), an extract of astragalus membranaceus (AM), is an effective cardioprotectant. Currently, little is known about the detailed mechanisms underlying cardioprotective effects of APS. The aims of this study were to investigate the potential effects and mechanisms of APS on apoptosis employing a model of high glucose induction of apoptosis in H9C2 cells.

METHODS

A model of high glucose induction of H9C2 cell apoptosis was adopted in this research. The cell viabilities were analyzed by MTT assay, and the apoptotic response was quantified by flow cytometry. The expression levels of the apoptosis related proteins were determined by Real-time PCR and western blotting.

RESULTS

Incubation of H9C2 cells with various concentrations of glucose (i.e., 5.5, 12.5, 25, 33 and 44 mmol/L) for 24 h revealed that cell viability was reduced by high glucose dose-dependently. Pretreatment of cells with APS could inhibit high glucose-induced H9C2 cell apoptosis by decreasing the expressions of caspases and the release of cytochrome C from mitochondria to cytoplasm. Further experiments also showed that APS could modulate the ratio of Bcl-2 to Bax in mitochondria.

CONCLUSIONS

APS decreases high glucose-induced H9C2 cell apoptosis by inhibiting the expression of pro-apoptotic proteins of both the extrinsic and intrinsic pathways and modulating the ratio of Bcl-2 to Bax in mitochondria.

Astragali Radix (AR), prepared from the roots of Astragalus membranaceus (FISCH. ex LINK) BUNGE or its variey, A. membranaceus (FISCH. ex LINK) BUNGE var. mongholicus (BUNGE) HSIAO., is one of the most used and valuable traditional Chinese medicines (TCMs). Historically, Hunyuan, Shanxi Province in China is the geo-authentic producing area of AR and crude AR from here called "geo-authentic." According to tradition, geo-authentic TCMs define both authenticity and quality. However, no scientific investigation has ever determined whether the superior quality of Hunyuan AR is due to the genetic characteristics or to the local environment. In our study, seeds of 30 AR samples representing the two varieties from different regions were cultivated in Hunyuan under the same conditions. A method, using ultra-performance liquid chromatography coupled with photodiode array detector and evaporative light scattering detectors, was developed to evaluate the quality through a simultaneous determination of four major isoflavonoids and four major saponins. The two AR varieties were successfully distinguished by principal component analysis while samples of the same species with different seeds origins could not be distinguished. A genetic study demonstrated that the internal transcribed spacer sequences of the nuclear ribosomal DNA in A. membranaceus var. mongholicus samples from different geographical regions were highly conservative. These results indicate that the content of active components in AR depends on the interaction of genotype and environment. At the varietal level, genetic properties appear to be more important for pharmaceutical quality than environmental factors, while on the intraspecific level environmental factors might be more important than genetic properties.

A novel polysaccharide named as AERP was extracted from industrial Astragalus membranaceus-extracted waste residue, which was composed of two components coded as AERP1 and AERP2. The structures of AERP1 and AERP2 were determined by HPLC-SEC-RID, HPLC-C18-UV, FT-IR, and NMR. The results showed that AERP1 was an acidic component with a molecular weight of 2.01 × 106 Da and glycosidic bonds of →3/5-α-araf-(1→, T-α-araf, →4,6-β-manp-(1→, →3/3,6-β-galp-(1→, →2/2,4-α-rha-(1→, →-4/4,6-α-glcp-(1→, →4-α-galpA-(1→ and →4)-6-OMe-α-galpA-(1→. AERP2 was a glucan with 2.11 × 103 Da by →4/6-α-glcp-(1→ linkage. In vitro, AERP retarded glucose diffusion significantly than each single component. In vivo, AERP had a hypoglycemic effect on db/db diabetic mice by alleviating the hyperglycemia, tissue impairment, and inhibiting cognitive impairment. AERP could alter the gut microbiota and modulate the composition of SCFAs. This study gives an opportunity for exploring the industrial waste of Astragalus membranaceus in diabetic complication therapy.

BACKGROUND

Danggui Buxue Tang (DBT), an herbal formula containing Angelica sinensis (AS) and Astragalus membranaceus (AM) (AS:AM = 1:5, designated as DBT1 here), has been used in Chinese medicine to enhance qi and blood circulation. In addition, DBT has served as a treatment for atopic dermatitis in dogs in Taiwan. It also may improve fibrosis in a rat model of pulmonary fibrosis.

OBJECTIVE

In this study, we evaluated the effect of oral administration of DBT1 in asthma in ovalbumin (OVA)-sensitized mice.

METHODS

Female BALB/c mice were sensitized and challenged with OVA and fed with DBT1 or modified formulas of DBT1, designated as DBT2 (AS:AM = 1:1) and DBT3 (AS:AM = 5:1), from days 21 to 27.

RESULTS

DBT1 suppressed airway hyperresponsiveness and eosinophil infiltration in bronchoalveolar lavage fluid (BALF) and lung, and Th2-associated cytokines and chemokines were inhibited in BALF. In addition, levels of OVA-immunoglobulin E (IgE) also were suppressed in serum. However, treatment with DBT2 or DBT3 showed no improved effects relative to DBT1 in treating asthmatic symptoms.

CONCLUSIONS

These results suggest that orally administered DBT (DBT1) can reduce allergic reactions in OVA-sensitized mice.

Radix Astragali (root of Astragalus; Huangqi) is a traditional Chinese medicine commonly used as an immunostimulant, hepatoprotective, diuretic, antidiabetic, analgesic, expectorant, and sedative drug. Although the species of Radix Astragali have been defined as Astragalus membranaceus and A. membranaceus var. mongholicus in Pharmacopoeia of China, their taxonomy remains controversial. The phylogenetic relationships among 10 Astragalus taxa, which are commonly found in China including A. membranaceus, A. membranaceus var. mongholicus, Astragalus propinquus, Astragalus lepsensis, Astragalus aksuensis, Astragalus hoantchy, Astragalus hoantchy subsp. dshimensis,Astragalus lehmannianus, Astragalus sieversianus, and Astragalus austrosibiricus, were determined using the DNA sequences of the 5S ribosomal RNA (5S rRNA) spacer, internal transcribed spacer region (ITS), and 18S rRNA coding region. The 5S rRNA spacer, ITS, and 18S rRNA, amplified by polymerase chain reaction from the isolated genomic DNAs, were sequenced. By using neighbor-joining and maximum parsimony analyses, phylogenetic trees were mapped by their sequence diversity. A. membranaceus and A. membranaceus var. mongholicus shared the greatest sequence homology. In addition, A. propinquus shared a closer relationship with A. membranaceus and A. membranaceus var. mongholicus, while other Astragalus species were less closely related. This is the first paper to show the phylogenetic relationship of Astragalus species related to Radix Astragali in China by the molecular genetic approach.

Formononetin is a naturally occurring isoflavone, which can be found in low concentrations in many dietary products, but the greatest sources of this substance are Astragalus membranaceus, Trifolium pratense, Glycyrrhiza glabra, and Pueraria lobata, which all belong to Fabaceae family. Due to its structural similarity to 17 β -estradiol, it can mimic estradiol's effect and therefore is considered as a "phytoestrogen." The aim of this study was to examine the effect of formononetin on mechanical properties and chemical composition of bones in rats with ovariectomy-induced osteoporosis. 12-week-old female rats were divided into 4 groups: sham-operated, ovariectomized, ovariectomized treated with estradiol (0.2 mg/kg) and ovariectomized treated with formononetin (10 mg/kg). Analyzed substances were administered orally for 4 weeks. Ovariectomy caused osteoporotic changes, which can be observed in bone biomechanical features (decrease of maximum load and fracture load and increase of displacements for maximum and fracture loads) and bone chemical composition (increase of water and organic fraction content, while a decrease of minerals takes place). Supplementation with formononetin resulted in slightly enhanced bone mechanical properties and bone chemistry improvement (significantly lower water content and insignificantly higher mineral fraction content). To summarize, administration of formononetin to ovariectomized rats shows beneficial effect on bone biomechanical features and chemistry; thus, it can prevent osteoporosis development.

Using a large-scale culture technique, the hairy roots of Astragalus membranaceus were produced with a yield reaching 10 g/L. The results from RP-HPLC detection showed that the contents of crude saponin and astragaloside i.v. in the hairy roots were 5.81% and 0.14%, respectively. Six isoflavonoid compounds were also determined. Polysaccharide analysis showed that the total polysaccharide content in the hairy roots was 22.97%; of this content, acidic 8.29% and soluble 14.88%. In comparison with the dry roots, the hairy roots contained higher crude saponin and soluble polysaccharide contents, similar astragaloside i.v. content and lower contents of 6 isoflavonoids, total and acidic polysaccharides, showing that the quality of both types of roots was similar. Regaining the immunity function of rats with low immunity after feeding the aqueous extract of the hairy roots produced by large-scale culture showed that its capacity was similar to the dry roots of A. membranaceus in increasing the immunity function. The results in this paper give evidence that the hairy roots may be a new source of A. membranaceus.

Dangguibohyul-tang (DBT), herbal formula composed of Astragalus membranaceus (AM) and Angelica sinensis (AS) at a ratio of 5 : 1, has been used for the treatment of various skin diseases in traditional medicine. We investigated the effect of DBT on allergic and inflammatory skin reaction in atopic dermatitis-like model compared to the single extract of AM or AS. DBT treatment showed the remission of clinical symptoms, including decreased skin thickness and scratching behavior, the total serum IgE level, and the number of mast cells compared to DNCB group as well as the single extract of AM- or AS-treated group. Levels of cytokines (IL-4, IL-6, IFN-γ, TNF-α, and IL-1β) and inflammatory mediators (NF-κB, phospho-IκBα, and phospho-MAPKs) were significantly decreased in AM, AS, and DBT groups. These results demonstrated that AM, AS, and DBT may have the therapeutic property on atopic dermatitis by inhibition of allergic and inflammatory mediators and DBT formula; a mixed extract of AM and AS based on the herb pairs theory especially might be more effective on antiallergic reaction as compared with the single extract of AM or AS.

It has been reported that Astragalus membranaceus, an Asian traditional herb, has an estrogenic effect in vitro. To examine the possible role of A. membranaceus extract with supplemental calcium (Ca) on bone status in calcium-deficient (LCa) ovariectomized (OVX) rats, a total of 48 female rats were divided into six groups: (1) normal control, (2) sham operation with LCa (sham-LCa), (3) OVX with LCa (OVX-LCa), (4) A. membranaceus supplementation with OVX-LCa (OVX-MLCa), (5) Ca supplementation with OVX (OVX-Ca), and (6) A. membranaceus and Ca supplementation with OVX (OVX-MCa). A. membranaceus ethanol extract (500 mg/kg BW) and/or Ca (800 mg/kg BW) were administered orally for 8 weeks along with a Ca-deficient diet. Results revealed that Ca supplementation with or without A. membranaceus extract significantly improved bone mineral density, biomechanical strength, and ash weight of the femur and tibia in OVX rats. High Ca with A. membranaceus combination supplementation significantly increased the ash weight of the femur and tibia and decreased urinary Ca excretion compared with supplementation of Ca alone. Uterine weight was not changed by A. membranaceus administration in OVX rats. These results suggest that A. membranaceus extract combined with supplemental Ca may be more protective against the Ca loss of bone than A. membranaceus or supplementation of Ca alone in calcium-insufficient postmenopausal women.

Astragali Radix is a widely and commonly used Chinese herbal medicine, which is derived from roots of Astragalus membranaceus var. mongholicus and Astragalus membranaceus. To find a quick and reliable method of distinguishing these two species of Astragali Radix and of determining the age of a sample, microscopic characteristics of the two species were compared using light microscopy. The results showed that the microscopic characteristics, such as number of layers of phellem, continuing lignified xylem bundles within spring wood and lignified parenchyma cells in the central part of the xylem could be used for the differentiation of the root of A. membranaceus from the root of A. membranaceus var. mongholicus. Growth rings (annual rings) were found for the first time in the roots of both species, and could determine the age of a sample. For the first time, radial fibers in both species of Astragali Radix and pipette-shaped fibers in A. membranaceus var. mongholicus were found. The structure of "rotten heart" cork tissue (decayed central xylem) and tubular cork tissue was carefully studied, and the arranged order of tissues in both "rotten heart" and tubular cork tissues is phelloderm and phellem from outside to inside, which is contrary to that in the periderm.

Twenty-four Mesorhizobium strains were isolated from desert soils in the Xinjiang region of China and were characterized by a polyphasic approach. These strains grouped into three clusters in IGS-RFLP, SDS-PAGE analysis of whole-cell proteins and BOX-PCR analysis, corresponding to genomic species V, VI and VII as found in a previous study. The results were supported by sequencing analyses of rrs, IGS, atpD and recA genes. Genospecies VII was most related to Mesorhizobium septentrionale, while genospecies V and VI were both most closely related to Mesorhizobium tianshanense, but were distinct from each other and from M. tianshanense. The DNA-DNA hybridization value between the representative strain CCBAU 83284 (genospecies VII) and the type strain of M. septentrionale was 90.1 %. Genospecies VII was thus defined as M. septentrionale. The DNA-DNA relatedness value for representative strains of genospecies V or VI with the related reference strains of recognized species were always lower than 60 %. Low values of DNA-DNA hybridization (32.79 %) between representative strains of genospecies V (CCBAU 83330(T)) and of VI (CCBAU 83306(T)) were also observed. Based upon these results, two novel species are proposed: Mesorhizobium gobiense sp. nov. represented by genospecies V (type strain, CCBAU 83330(T)=LMG 23949(T)=HAMBI 2974(T)) and Mesorhizobium tarimense sp. nov. represented by genospecies VI (type strain, CCBAU 83306(T)=LMG 24338(T)=HAMBI 2973(T)). Strain CCBAU 83278 grouped as the most peripheral member with genospecies VI in SDS-PAGE of whole-cell proteins and BOX-PCR analysis and in the phylogenetic tree of 16S-23S rRNA intergenic spacer (IGS) sequences. The results of analyses of rrs, atpD and recA gene sequences, as well as those of DNA-DNA hybridization studies, strongly supported the suggestion that this strain belonged to a species quite different from genospecies V and VI and from any other recognized species of the genus Mesorhizobium. As only one strain has been isolated to date, strain CCBAU 83278 was not proposed as a novel species in this study. Mesorhizobium gobiense sp. nov. and Mesorhizobium tarimense sp. nov. could be differentiated from each other as well as from recognized species of the genus Mesorhizobium on the basis of phenotypic characteristics. The symbiotic loci (nodC and nifH) of the two novel species formed two phylogenetic branches related to Mesorhizobium loti and M. tianshanense. The type strains of the two novel species were able to nodulate Glycyrrhiza uralensis, Lotus corniculatus, Oxytropis glabra and Robinia pseudoacacia but not Astragalus membranaceus, Leucaena leucocephala, Phaseolus vulgaris, Pisum sativum or Medicago sativa.

BACKGROUND

Recent studies have revealed the important role of free radicals in renal damage induced by high-energy shock waves (HESW). This study aimed at investigating the effects of Astragalus membranaceus, a traditional Chinese medicinal herb, on free radical-mediated HESW-induced damage to renal tubules in a live rabbit model.

METHODS

Forty-five healthy male New Zealand white rabbits were randomly divided into three groups: control group (n = 15), sham group (n = 15), and herb-treated group (n = 15). Three days prior to HESW application, the controls received verapamil (0.4 mg/kg), the shams received physiological saline (20 ml), and the herb-treated animals received Astragalus membranaceus (2.4 g/kg) intravenously. HESW (1500 shocks, 18 kV) was applied to the right kidneys of all anesthetized rabbits. We measured superoxide dismutase (SOD) and malondialdehyde (MDA) levels before and after shock treatment in blood and kidney homogenates. Histopathological changes were also observed.

RESULTS

MDA levels increased and SOD activity decreased significantly in the sham group (P < 0.05 for both) after shock treatment. MDA levels showed a much less increase in the controls (P < 0.05) and did not increase to statistically significant levels in the group receiving Astragalus membranaceus (P>> 0.05). SOD values were significantly higher in the controls than in the shams (P < 0.05). By contrast, SOD levels recovered rapidly in the rabbits receiving Astragalus membranaceus, reaching a nadir within 24 hours, and returning to baseline more quickly than in control and sham rabbits (P < 0.05). Histopathological examinations showed that renal tubular damage in the controls was less severe than in the shams, while damage in the Astragalus membranaceus group was even more mild, with rapid recovery in comparison with the controls.

CONCLUSIONS

This study provides preliminary evidence indicating that Astragalus membranaceus has strong protective effects on free radical-mediated renal tubular damage induced by HESW and that these effects are superior to the effects of verapamil.

We have extracted and roughly purified astragalosides (AS) from Astragalus membranaceus, a natural herb used as a traditional Chinese medicine, regarded to have pharmacodynamic benefits of protecting injured myocardium. We hypothesized that the astragalosides might exert beneficial effect in myocardial lesion by preserving both energy metabolism and Ca(2+) homeostasis. Sprague-Dauley (SD) rats were injected with isoproterenol (ISO) subcutaneous (s.c.) at a dose of 5 mg/kg/day consecutively for two days as models and were treated with astragalosides and trimetazidine intraperitoneally (i.p.) respectively, at a dose of 5 mg/kg/day one day prior to isoproterenol for 8 days. The histological changes were alleviated in isoproterenol-injected SD rats treated with astragalosides. Compared with isoproterenol-injected rats, the concentration of myocardial intracellular [Ca(2+)]i was decreased, L-type Ca(2+) current density and sarcoplasmic reticulum (SR) Ca(2+) load were recovered, the concentration of myocardial ATP was increased and phosphocreatine (PCr) was decreased in rats treated with astragalosides. In conclusion, the efficacious treatment of astragalosides for myocardial injury might be through regulating intracellular Ca(2+) homeostasis and energy metabolism.

Astragaloside IV (AS-IV) is one of the main active constituents of Astragalus membranaceus, which has various actions on the cardiovascular system. However, its electrophysiological mechanisms are not clear. In the present study, we investigated the effects of AS-IV on action potentials and membrane currents using the whole-cell patch clamp technique in isolated guinea-pig ventricular myocytes. AS-IV prolonged the action potential duration (APD) at all three tested concentrations. The peak effect was achieved with 1×10(-6) M, at which concentration AS-IV significantly prolonged the APD at 95% repolarization from 313.1±38.9 to 785.3±83.7 ms. AS-IV at 1×10(-6) M also enhanced the inward rectifier K(+) currents (I(K1)) and inhibited the delayed rectifier K(+) currents (I(K)). AS-IV (1×10(-6) M) strongly depressed the peak of voltage-dependent Ca(2+) channel current (I(CaL)) from -607.3±37.5 to -321.1±38.3 pA. However, AS-IV was not found to affect the Na(+) currents. Taken together, AS-IV prolonged APD of guinea-pig ventricular myocytes, which might be explained by its inhibition of I(K). AS-IV also influences Ca(2+) signaling through suppressing ICaL.

Exposure to ultraviolet (UV) light reduces levels of type I collagen in the dermis and results in human skin damage and premature skin aging (photoaging). This leads to a wrinkled appearance through the inhibition of transforming growth factor‑β (TGF‑β)/Smad signaling. UV irradiation increases type I collagen degradation through upregulating matrix metalloproteinase (MMP) expression. Astragaloside IV (AST) is one of the major active components extracted from Astragalus membranaceus. However, its multiple anti‑photoaging effects remain to be elucidated. In the present study, the effects of AST against collagen reduction in UV‑induced skin aging in human skin fibroblasts were investigated. The expression of type I procollagen (COL1), MMP‑1, TGF‑βRⅡ and Smad7 were determined using reverse transcription‑polymerase chain reaction, western blotting and ELISA, respectively. UV irradiation inhibits type I collagen production by suppressing the TGF‑β/Smad signaling pathway and increasing COL1 degradation by inducing MMP‑1 expression. Transforming growth factor‑β type II protein and COL1 mRNA decreased but MMP‑1 and Smad7 levels increased in the photoaging model group, which was reversed by topical application of AST. AST prevents collagen reduction from UV irradiation in photoaging skin by improving TGF‑β/Smad signaling suppression and inhibiting MMP‑1, thus AST may be a potential agent against skin photoaging.

Diabetic nephropathy (DN) is one of the most important causes of end‑stage renal disease. Astragaloside IV (AS-IV) is a saponin isolated from Astragalus membranaceus, which possesses various pharmacological activities. AS‑IV prevents podocyte apoptosis and ameliorates renal injury in DN; however, few studies have focused on its effects on ion channels. The transient receptor potential channel 6 (TRPC6) is an important Ca2+‑permeable ion channel in podocytes, which is involved in high glucose (HG)-induced podocyte apoptosis. The aim of the present study was to investigate whether AS‑IV prevented HG‑induced podocyte apoptosis via TRPC6. Cultured podocytes were pre‑treated with 10, 20 or 40 µM AS‑IV for 1 h prior to HG exposure for 24 h. Apoptosis, cell viability, expression of TRPC6, nuclear factor of activated T cells (NFAT2) and B‑cell lymphoma 2‑associated X protein (Bax), as well as the intracellular Ca2+ concentration were subsequently analyzed. The results indicated that HG induced podocyte apoptosis and upregulation of TRPC6, and increased intracellular Ca2+. Furthermore, enhanced NFAT2 and Bax expression was detected. Conversely, AS‑IV protected HG‑induced podocyte apoptosis, downregulated TRPC6 expression and suppressed intracellular Ca2+ in HG-stimulated podocytes. AS‑IV also suppressed NFAT2 and Bax expression. These results suggest that AS‑IV may prevent HG-induced podocyte apoptosis via downregulation of TRPC6, which is possibly mediated via the calcineurin/NFAT signaling pathway.

Han-Dan-Gan-Le (HDGL), a Chinese herb preparation composed of Stephaniat tetrandra, Salvia miltorrhiza, Radix paeoniae, Astragalus membranaceus, and Ginkgo biloba, has been used to treat human liver fibrosis. This study was designed to examine the therapeutic effect of HDGL on chemical-induced liver fibrosis in adult Wistar rats. Liver fibrosis was produced in rats by carbon tetrachloride (1.2 ml CCl(4)/kg, 2 times/week, after an initial dose of 5.0 ml CCl(4)/kg, sc), plus a diet of 20% fat, 0.05% cholesterol (continuous) and 30% alcohol in the drinking water ad libitum (every other day) for 8 weeks. HDGL (0.5 and 1.0 g/kg, ig, daily for 6 weeks) was administered to rats 72 hrs after the last dose of CCl(4) to examine its therapeutic effects on chemical-induced liver fibrosis. Upon pathological examination, the HDGL treatment had significantly reversed chemical-induced liver fibrosis and other hepatic lesions. Hepatic collagen accumulation induced by CCl(4) was markedly reduced by HDGL treatment, as evidenced by hepatic collagen content and by immunohistochemical analysis of type-I collagen in liver. HDGL appeared to stimulate the collagenolytic process in the liver, as a 30-50% increase in urinary excretion of hydroxyproline was observed with HDGL treatment as compared to rats only given CCl(4). In conclusion, HDGL can effectively reverse chemically induced liver fibrosis, and this appears to be due, at least in part, to the stimulation of hepatic collagenolysis, resulting in a resolution of hepatic fibrosis.

Anthracyclines are antitumor antibiotics with significant activity against solid and hematologic malignancies. One problem preventing more widespread use has been the development of cardiotoxicity. To determine whether antioxidant agents can reduce the cardiotoxicity of anthracyclines, a herb Astragalus membranaceus was introduced, which has been widely used for the treatment of cardiovascular diseases in China and was confirmed to be an effective antioxidant agent recently. Pre-treatment with Astragalus membranaceus significantly attenuated the daunorubicin-induced increases of reactive oxygen species (p < 0.001), apoptosis (p < 0.05) and the secretions of LDH (p < 0.01) in cultured neonatal cardiomyocytes. Astragalus membranaceus also raised the EC(50) of daunorubicin 1.24-fold. Compared with Astragalus membranaceus, N-acetyl-L-cysteine had similar effects on daunorubicin-induced cell injury, however, superoxide dismutase reduced reactive oxygen species without attenuating apoptosis. The subcellular distribution of DNR was similar to the distribution of MitoTracker Red 580 in mitochondria, which was mainly in the cytoplasm around the nuclear membrane in cultured neonatal cardiomyocytes. In conclusion, the results suggested that Astragalus membranaceus is potentially protective against daunorubicin cardiotoxicity by decreasing free radical release and apoptosis in cultured neonatal cardiomyocytes. The main subcellular distribution of daunorubicin may be in the mitochondria.

The aim of this study was to investigate the mechanism underlying the effects of Astragalus membranaceus injection (AMI) on myelopoiesis in myelosuppressed mice. At 72 h after cyclophosphamide injection (250 mg/kg), the mice were administered AMI (500 mg/kg) intraperitoneally for 6 consecutive days or an equivalent volume of saline as a control. Murine colony-forming unit-fibroblast (CFU-F) formation, production of IL-6 and GM-CSF by bone marrow stromal cells (BMSC), and bcl-2 protein and mRNA expression in BMSC were measured by CFU-F assay, ELISA, immunohistochemistry and in situ hybridization. The results indicated that AMI improved the hematopoietic microenvironment by enhancing the BMSC survival and proliferation of CFU-F, production of IL-6 as well as GM-CSF by BMSC and bcl-2 protein and mRNA expression in BMSC, which promoted myelopoiesis. The data may provide a mechanistic basis for applying this ancient Chinese herb to promote hematopoiesis as an efficacious adjuvant therapy against myelosuppression induced by anti-cancer therapy.

Roots of Astragalus membranaceus (Fish.) Bge. var. mongholicus (Bge.) Hsiao (A. membranaceus) have been long used as an auxiliary reagent supporting cancer treatment. Here, we compared the chemical composition and antitumor immunomodulating activity of polysaccharides from roots of A. membranaceus (PAMs) from five major habitats in Inner Mongolia, PR China. We revealed that compositions of monosaccharides and amino acids were comparable among PAMs from different habitats. However, amounts of selenium varied widely in roots of A. membranaceus and PAMs. PAMs selenium-dependently repressed the in vivo proliferation of transplanted H22 ascitic hepatoma and S180 sarcoma cells with low toxic impacts on tumor-bearing mice. Selenium-containing PAMs ameliorated host CD4+ T cell apoptosis and serum cytokine dysregulation induced by tumor transplantation, leading to the enhancement of cytotoxic activities of natural killer and CD8+ T cells. Moreover, PAMs also selenium-dependently improved the phagocytotic function of intra-abdominal macrophages and suppressed M2-like polarization of tumor-associated macrophages. These data suggested that the selenium content varies in the roots of A. membranaceus and PAMs from different geographical origins dramatically and selenium is an important contributor to the antitumor immunomodulation activities of PAMs.

Tumors are known to produce factors suppressing immune functions. We previously showed that a murine renal cell carcinoma (Renca) suppressed macrophage function in vitro and that this suppression was abolished by co-incubation with extracts of two Chinese medicinal herbs. We now report that these phytochemicals are capable of inhibiting growth of Renca in vivo. BALB/c mice were transplanted intraperitoneally (IP) with 1-2 x 10(5) Renca cells. One day after tumor transplant, mice were randomized into two groups. One group was treated IP, daily for 10 days, with 100 microliters of phytochemicals containing 500 micrograms each of Astragalus membranaceus and Ligustrum lucidum, while the other group received saline as controls. A cure rate of 57% was obtained with these phytochemicals when the initial tumor load was 2 x 10(5), and 100% when the initial tumor load was 1 x 10(5). Additional experiments were performed to investigate the mechanisms involved in this protection. Splenic macrophages from tumor-bearing mice were shown to have depressed chemiluminescent oxidative burst activity, and this depression was restored with phytochemical treatment. Splenocytes from mice transplanted with Renca responded less favorably to interleukin-2 (IL-2) in generating lymphokine-activated killer (LAK) cells; again this depression was restored with phytochemical treatment. Our data suggest that these phytochemicals may have exerted their antitumor effects via augmentation of phagocyte and LAK cell activities.