Purification of triterpenoid saponins and 25R/25S-inokosterone from Achyranthes bidentata Bl. by high-speed countercurrent chromatography coupled silver nitrate coordination.

An effective method by high-speed countercurrent chromatography coordinated with silver nitrate for the preparative separation of sterones and triterpenoid saponins from Achyranthes bidentata Blume was developed. Methyl tert-butyl ether/n-butanol/acetonitrile/water (4:2:3:8, v/v/v/v) was selected for 20-hydroxyecdysone (compound 1), chikusetsusaponin IVa methyl ester (compound 4), 2'-glycan-11-keto-pigmented saponin V (compound 5), as well as a pair of isomers of 25S-inokosterone (compound 2) and 25R-inokosterone (compound 3), which were further purified by silver nitrate coordinated high-speed countercurrent chromatography. What is more, dichloromethane/methanol/isopropanol/water (6:6:1:4, v/v/v/v) was applied for calenduloside E (compound 6), 3ß-[(O-ß-d-glucuronopyranosyl)-oxy]-oleana-11,13-dien-28-oic acid (compound 7), zingibroside R1 (compound 8) and chikusetsusaponin IVa (compound 9). Adding Ag+ to the solvent system resulted in unique selectivity for 25R/25S isomers of inokosterone, which increased the complexing capability and stability of Ag+ coordinated 25S-inokosterone, as well as the α value between them. These results were further confirmed by the computational calculation of geometry optimization and frontier molecular orbitals assay. Comprehensive mass spectrometry and nuclear magnetic resonance analysis demonstrated the structures of the obtained compounds.

Chemometric guided isolation of new triterpenoid saponins as acetylcholinesterase inhibitors from seeds of Achyranthes bidentata Blume.

Achyranthes bidentata Blume (Amaranthaceae) is an annual or perennial herb widely used as ethnomedicine in Traditional Chinese Medicine for treating fever, cold, ulcers, mensural pain, dementia, and osteoporosis. In the current study, UPLC-IM-Q-TOF-MS/MS-based chemometric approach was adopted for the tentative identification of fifty-six compounds in the extract and fractions of A.bidentata seeds. Further, the chemometric-guided isolation led to the isolation of two previously undescribed oleanane-type triterpenoid saponins, named achyranosides A-B (27 and 30), along with three known compounds (31, 44, and 23) from water fraction of A. bidentata seeds. The structures of new compounds were elucidated based on the detailed analysis of NMR, HR-ESI-MS, FT-IR spectral data, and GC-FID techniques. The isolated compounds in vitro acetylcholinesterase inhibitory activity revealed the promising activity of chikusetsusaponin IVa (23) (IC50 = 63.7 µM) with mixed type of AChE inhibition in enzyme kinetic studies. Additionally, in silico binding free energy of isolated compounds disclosed the greater stability of enzyme-ligand complex owing to underlying multiple H-bond interactions. Overall, the study demonstrates the effectiveness of a chemometric-guided approach for the phytochemical exploration and isolation of new oleanane-type triterpenoid saponins from A. bidentata seeds.

Application of microscopy and spectroscopy in investigating anti-cancer potential of Achyranthes aspera L. leaves.

The genus Achyranthes belong to the family Amaranthaceae which constitutes an important group of herbs and shrubs with immense medicinal value. The present research work was conducted to investigate the anticancer potential of Achyranthes aspera L. leaves by focusing on the antioxidant, aniproliferative and antimitotic activities of leaf extracts. Plant extraction was carried out by soxhelt method with different solvents. Phytochemical characterization of the plants extracts using chemical methods identified the presence of cardiac glycosides, saponins, coumarins, proteins, tannins, flavonoids and triterpenes. Alkaloid was present in methanolic and ethanolic extract. High performance liquid chromatography showed presence of different concentration of myricetin, quercetin and kaempferol in different extracts with the highest concentration of myricetin (84.53 µg/mL) in n-butanolic extract. The extracts were then tested for antioxidant activity using 2,2-diphenylpicrylhydrazyl (DPPH) radical scavenging assay by spectrophotometric method. In DPPH radical scavenging assay, antioxidant activity of A. aspera ranged between 79.78 ± 0.034% and 58.63 ± 0.069%. Highest antioxidant activity was observed for methanolic extract and lowest for acetone. Antimitotic activity was determined by using Allium cepa assay in which microscopic investigation was carried out to observe normal and abnormal phases of mitosis. In this assay, n-butanolic extract had highest antimitotic activity with minimum mitotic index at 2 mg/mL (57 ± 0.0351%). The plant extracts also caused chromosomal and mitotic aberrations which were clearly observed under 40× and 100× magnification of compound microscope. Antiproliferative activity was determined by using yeast cell model in which light microscope with hemocytometer was used for cell counting. In case of Antiproliferative activity, the ethyl acetate extract of A. aspera had highest antiproliferative activity with lowest cell viability (22.14 ± 0.076%) at highest extract concentration (2 mg/mL) while methanol extract of A. aspera had highest antiproliferative activity with lower cell viability (24.24 ± 0.057%) at lowest extract concentration (0.25 mg/mL). The results of the study indicated that the leaves extract of A. aspera have strong potential to be used as a source of anti-cancer agent. RESEARCH HIGHLIGHTS: Achyranthes aspera L. leaves have various phytochemicals which contribute to its medicinal properties Various extracts of the leaves of A. aspera L. possess antioxidant, antimitotic and antiproliferative potential The results of the study indicated that the leaves extract of A. aspera have strong potential to be used as a source of anti-cancer agent.

Oligo/polysaccharides from Cyathula officinalis and Achyranthes bidentata: a review of structures and bioactivities.

OBJECTIVES: Oligo-/polysaccharides from Cyathula officinalis Kuan (COPs) and Achyranthes bidentata Blume (ABPs) have attracted researchers' attention in the fields of healthy food supplements and traditional Chinese medicine (Niúxi) due to their multiple bioactivities combined with their nontoxic and highly biocompatible nature. The purpose of this paper was to provide a systematic and comprehensive overview of the extraction, purification, and structural analysis methods, chemical characteristics, biological activities, and structure bioactivity relationship. Furthermore, the possible development trends and perspectives for future research, and traditional uses of Niúxi are also summarized. METHODS: All the information was gathered from a library search and scientific databases. KEY FINDINGS: Although COPs and ABPs are derived from different plants, they have similar structural features in type, structure, and glycosidic linkage patterns and biological activities in vivo and in vitro. However, there are differences in monosaccharide compositions, which can be used as an identification mark. CONCLUSIONS: As traditional Chinese herbal medicine, C. officinalis and A. bidentata have similar pharmacological activities. The COPs and ABP possess wide pharmacological effects such as antitumor, antioxidant, anti-osteoporosis, and anti-inflammatory. Meanwhile, the biological activity and structure-activity relationship of purified COPs and ABPs are less studied, future research should focus on them.

Experimental and pharmacoinformatic approaches unveil the neuropharmacological and analgesic potential of chloroform fraction of Roktoshirinchi (Achyranthes ferruginea Roxb.).

ETHNOPHARMACOLOGICAL RELEVANCE: Achyranthes ferruginea (A. ferruginea) Roxb. is a common plant used in traditional medicine in Asia and Africa. It has a variety of local names, including "Gulmanci" in Nigeria, "Dangar" in Pakistan, "Thola" in Ethiopia, and "Roktoshirinchi" in Bangladesh. It is edible and has several ethnomedical uses for a wide range of illnesses, including hysteria, dropsy, constipation, piles, boils, asthma, and shigellosis. However, the neuropharmacological and analgesic potential of A. ferruginea remains uninvestigated. AIM OF THE STUDY: To assess the neuropharmacological and analgesic potential of A. ferruginea through a multifaceted approach encompassing both experimental and computational models. MATERIALS AND METHODS: Methanol was used to extract the leaves of A. ferruginea. It was then fractionated with low to high polar solvents (n-hexane, chloroform, ethyl acetate, and water) to get different fractions, including chloroform fraction (CLF). The study selected CLF at different doses and conducted advanced chemical element and proximate analyses, as well as phytochemical profiling using GC-MS. Toxicological studies were done at 300 µg per rat per day for 14 days. Cholinesterase inhibitory potential was checked using an in-vitro colorimetric assay. Acetic acid-induced writhing (AAWT) and formalin-induced licking tests (FILT) were used to assess anti-nociceptive effects. The forced swim test (FST), tail suspension test (TST), elevated plus maze (EPM), hole board test (HBT), and light and dark box test (LDB) were among the behavioral tests used to assess depression and anxiolytic activity. Network pharmacology-based analysis was performed on selected compounds using the search tool for interacting chemicals-5 (STITCH 5), Swiss target prediction tool, and search tool for the retrieval of interacting genes and proteins (STRING) database to link their role with genes involved in neurological disorders through gene ontology and reactome analysis. RESULTS: Qualitative chemical element analysis revealed the presence of 15 elements, including Na, K, Ca, Mg, P, and Zn. The moisture content, ash value, and organic matter were found to be 11.12, 11.03, and 88.97%, respectively. GC-MS data revealed that the CLF possesses 25 phytoconstituents. Toxicological studies suggested the CLF has no effects on normal growth, hematological and biochemical parameters, or cellular organs after 14 days at 300 µg per rat. The CLF markedly reduced the activity of both acetylcholinesterase and butyrylcholinesterase (IC50: 56.22 and 13.22 µg/mL, respectively). Promising dose-dependent analgesic activity (p < 0.05) was observed in chemically-induced pain models. The TST and FST showed a dose-dependent substantial reduction in immobility time due to the CLF. Treatment with CLF notably increased the number of open arm entries and time spent in the EPM test at doses of 200 and 400 mg/kg b.w. The CLF showed significant anxiolytic activity at 200 mg/kg b.w. in the HBT test, whereas a similar activity was observed at 400 mg/kg b.w. in the EPM test. A notable increase in the amount of time spent in the light compartment was observed in the LDB test by mice treated with CLF, suggesting an anxiolytic effect. A network pharmacology study demonstrated the relationship between the phytochemicals and a number of targets, such as PPARA, PPARG, CHRM1, and HTR2, which are connected to the shown bioactivities. CONCLUSIONS: This study demonstrated the safety of A. ferruginea and its efficacy in attenuating cholinesterase inhibitory activity, central and peripheral pain, anxiety, and depression, warranting further exploration of its therapeutic potential.

Isolation of Calenduloside E from Achyranthes bidentata Blume and its effects on LPS/D-GalN-induced acute liver injury in mice by regulating the AMPK-SIRT3 signaling pathway.

BACKGROUND: Acute liver injury (ALI) is a frequent fatal liver disease with a high mortality. Calenduloside E (CE) is a pentacyclic triterpenoid derived from Achyranthes bidentata Blume. It has been found that liver injury is associated with mitochondrial dysfunction, and activation of the AMPK-SIRT3 signaling pathway protects the mitochondrial function to play a role in resistance to the disease. However, whether CE is protective against ALI through the AMPK-SIRT3 signaling pathway is unclear. PURPOSE: To clarify the influences of Calenduloside E (CE) isolated from Achyranthes bidentata Blume on LPS/D-GalN-induced Acute liver injury (ALI). METHODS: A mouse model of ALI was developed, intraperitoneal injection of 10 µg/kg LPS and 700 mg/kg D-GalN, histopathological, oxidative stress, and immune inflammation of the mice were monitored. The mechanism of CE influencing liver injury was investigated by examining the gut microbiota, mitochondrial dysfunction, and the AMPK-SIRT3 signaling pathway. The antagonistic effects of specific AMPK and SIRT3 blocker, as well as AMPKα1, AMPKα2, SIRT3 transfection-mediated silencing were investigated to confirm the role of the AMPK-SIRT3 signaling pathway in this process. RESULTS: CE relieved liver pathological damage of mice and led to reduced oxidative stress and immune inflammation in mice, affected the balance of gut microbiota in mice with liver injury, as well as energy metabolism, and regulated mRNA and protein expressions of AMPK-SIRT3 signaling pathway. In addition, in vitro studies showed that CE relieved mitochondrial respiratory and protein expressions of AMPK-SIRT3 signaling pathway in LPS/D-GalN-induced AML12 and LX2 cells, and such effect was blocked by AMPK and SIRT3 inhibitors. Furthermore, silencing of AMPKα1, AMPKα2, and SIRT3 blocked the effects of CE. Overall, the influences of CE on mice with liver injury is tuned by the AMPK-SIRT3 signaling pathway. CONCLUSION: CE mediates mitochondrial function and eventually regulate energy metabolism by regulating the AMPK-SIRT3 signaling pathway. The results of this study provide molecular evidences for application of CE in treatment of ALI and provide references to the drug development for ALI.

Deep chemical identification of phytoecdysteroids in Achyranthes bidentata Blume by UHPLC coupled with linear ion trap-Orbitrap mass spectrometry and targeted isolation / 中国天然药物

Achyranthes bidentata Blume is widely used as a traditional Chinese medicine with the effects of nourishing the liver and kidneys and strengthening muscles and bones. In this work, a rapid and simple strategy was developed for characterizing phytoecdysteroids by ultra-high-performance liquid chromatography coupled with liner ion trap-Orbitrap mass spectrometry using electrospray ionization in the negative mode. As a result, 47 phytoecdysteroids were unambiguously or tentatively characterized. Among them, seven known compounds were identified according to the reference standards along with molecular formula, retention time and fragmentation patterns, while others were mostly potential new compounds. Through targeted isolation, the structures of three new compounds were determined by NMR spectra, which were consistent with LC-MS characterization. The present study provides an efficient method to deeply characterize phytoecdysteroids.

Pharmacokinetics of Achyranthes bidentata on adjuvant arthritis rats by microdialysis and UHPLC-MS/MS / 中国中药杂志

To investigate the " drug-guide" effect of Achyranthes bidentata saponins( ABS) and geniposide( GE) in the treatment on adjuvant arthritis( AA) rats. A UHPLC-MS/MS method for the quantitative determination of GE,zingibroside R1,ginsenoside Ro and chikusetsu saponin Ⅳa in rat blood and joint dialysate was established. After single or combined administration with ABS and GE was given to AA rat model,a microdialysis sampling method for rat joint cavity and jugular vein blood vessels was established to collect microdialysis samples. Waters Acquity HSS C_(18) column was used to separate the above four components,with mobile phase as acetonitrile-0. 1% formic acid water as mobile phase for gradient elution. ESI source was adopted for mass spectra in a negative ion scanning mode. Multiple reaction monitoring( MRM) mode was applied to detect the above four components. The methodological results showed that GE,zingibroside R1,ginsenoside Ro and chikusetsu saponin Ⅳa demonstrated a good linear relationship within the concentration ranges of 2-4 000,16-4 096,14-3 584,23-5 888 μg·L-1 respectively. The precision,accuracy,stability and matrix effect of these four ingredients reached the requirements of quantitative analysis of biological samples. The pharmacokinetic results demonstrated that the combined administration of ABS and GE( 60 mg·kg~(-1)+60 mg·kg~(-1)) can increase the degree of GE in joint cavity distribution,and the AUCjoint/AUCplasmwere twice of that of single administration of GE( 60 mg·kg~(-1)),which indicated that ABS might played a vital role in GE's distribution to joint cavity. Moreover,there was no significant difference between the distribution trend of total three ABS and GE in rats. The pharmacodynamics results showed that the combined administration of ABS and GE has stronger effects on paw swelling,arthritis index and synovial pathomorphology of AA rats than single administration of GE,which suggested that ABS might improve GE's anti-inflammatory effect in AA rats. Based on the above results,ABS has a targeting effect in increasing GE's concentration in joint cavity,with a synergy in efficacy.

Studies on effects of Achyranthes bidentata on tongsaimai pellets main active ingredients chlorogenic acid, isoliquiritin, harpagoside and glycyrrhizin in vivo pharmacokinetics / 中国中药杂志

To study on the effects of Achyranthes bidentata on Tongsaimai pellets main active ingredients chlorogenic acid, isoliquiritin, harpagoside and glycyrrhizin in rats in vivo pharmacokinetic behaviors, a method for the simultaneous determination of chlorogenic acid, isoliquiritin, harpagoside and liquiritigenin in rat plasma was established by UPLC-MS/MS. The analysis was performed on a waters Acquity BEH C18 column (2.1 mm x 100 mm, 1.7 microm) with the mixture of acetonitrile and 0.1% formic acid/water as mobile phase, and the gradient elution at a flow rate of 0.3 mL x min(-1). The analytes were detected by tandem mass spectrometry with the electrospray ionization (ESI) source and in the multiple reaction monitoring (MRM) mode. It turned out that the analytes of Tongsaimai pellets groups C(max) and AUC(Q-infinity) values were higher than that with A. bidentata group, and the C(max) values of chlorogenic acid had significantly difference (P < 0.05), the AUC(0-infinity) values of chlorogenic acid and glycyrrhizin had significantly difference (P < 0.05); The T(max) and CL values of two groups had no significantly difference. Results showed that the established method was specific, rapid, accurate and sensitive for the studies of Tongsaimai pellets four main active ingredients in rat in vivo pharmacokinetic, and A. bidentata have varying degrees of effects on Tongsaimai pellets four main active ingredients in rat in vivo pharmacokinetic behaviors.

Effect of Achyranthes bidentata polysaccharides stimulated dendritic cells co-cultured with cytokine induced killer cells against SW480 cells / 中国中药杂志

Achyranthes bidentata polysaccharides (ABPS) was extracted from the root of A. bidentata. Dendritic cells (DC), which were stimulated with ABPS and/or tumor antigen SW480, were co-cultured with cytokine induced killer cells (CIK) to test the cytotoxic effect on colon cancer cell line SW480. Peripheral blood mononuclear cells (PBMNCs) which were separated from human peripheral blood were cultured to DC and CIK separately. (1) DC were divided into four groups: pure DC served as control group; ABPS (50 mg x L(-1)) stimulated DC served as experimental group; SW480 tumor antigen stimulated DC served as the second experimental group; ABPS (50 mg x L(-1)) and SW480 tumor antigen co-stimulated DC served as the third experimental group. Flow cytometry was used to detect the difference of the positive rate of molecules in the cell surface of DC, include CD80, CD86, CD1c, CD40, HLA-DR (6 samples for each group). (2) The four DC groups were mixed with CIK at the ratio 1:5 and acted as effect cells (DC + CIK groups), and the colon cancer cell line SW480 acted as target cells. The effect cells and the target cells were mixed together at the ratio 30: 1, 20:1 and 10:1 separately, and the CCK-8 kit was used to test the cytotoxic effect on colon cancer cell line SW480. (3) At the mixing ratio 30:1 of effect cells and target cells, ELISA was used to test the level of cytokines secretion, including IL-2, IL-12p70, IL-17 and TNF-alpha, in the liquid supernatant of every test group (3 duplication per sample). The results showed as following: (1) The positive rates of CD80, CD11c, HLA-DR, in the cell surface of DC which was co-stimulated by ABPS (50 mg x L(-1)) and SW480 tumor antigen, were obviously higher than the other DC groups (P < 0.05), and the positive rates of CD86, CD40 were obviously higher than the pure DC group (P < 0.05), and there was no remarkable difference with the other two DC groups. (2) At the mixing ratio 30:1, 20:1 and 10:1 of the effect cells and the target cells, the cytotoxic effect of ABPS stimulated DC + CIK group and SW480 tumor antigen stimulated DC + CIK group was obviously higher than DC + CIK group (P < 0.05), the cytotoxic effect of ABPS and SW480 tumor antigen co-stimulated DC + CIK group was obviously higher than all the other groups. (3) At the mixing ratio 30:1 of the effect cells and the target cells, the secretion levels of IL-12p70 and TNF-alpha in the liquid supernatant of the ABPS and SW480 tumor antigen co-stimulated DC + CIK group were obviously higher than all the other groups (P < 0.05), the secretion levels of IL-2 and IL-17 in the liquid supernatant of every test group have no remarkable difference. The cytotoxic effect of ABPS stimulated DC + CIK on SW480 was obviously increased. The cytotoxic effect of ABPS and SW480 tumor antigen co-stimulated DC + CIK group was obviously higher than all the other.

In vitro clonal propagation of Achyranthes aspera L. and Achyranthes bidentata Blume using nodal explants

<p><b>OBJECTIVE</b>To develop the reproducible in vitro propagation protocols for the medicinally important plants viz., Achyranthes aspera (A. aspera) L. and Achyranthes bidentata (A. bidentata) Blume using nodal segments as explants.</p><p><b>METHODS</b>Young shoots of A. aspera and A. bidentata were harvested and washed with running tap water and treated with 0.1% bavistin and rinsed twice with distilled water. Then the explants were surface sterilized with 0.1% (w/v) HgCl2 solutions for 1 min. After rinsing with sterile distilled water for 3-4 times, nodal segments were cut into smaller segments (1 cm) and used as the explants. The explants were placed horizontally as well as vertically on solid basal Murashige and Skoog (MS) medium supplemented with 3% sucrose, 0.6% (w/v) agar (Hi-Media, Mumbai) and different concentration and combination of 6-benzyl amino purine (BAP), kinetin (Kin), naphthalene acetic acid (NAA) and indole acetic acid (IAA) for direct regeneration.</p><p><b>RESULTS</b>Adventitious proliferation was obtained from A. aspera and A. bidentata nodal segments inoculated on MS basal medium with 3% sucrose and augmented with BAP and Kin with varied frequency. MS medium augmented with 3.0 mg/L of BAP showed the highest percentage (93.60±0.71) of shootlets formation for A. aspera and (94.70±0.53) percentages for A. bidentata. Maximum number of shoots/explants (10.60±0.36) for A. aspera and (9.50±0.56) for A. bidentata was observed in MS medium fortified with 5.0 mg/L of BAP. For A. aspera, maximum mean length (5.50±0.34) of shootlets was obtained in MS medium augmented with 3.0 mg/L of Kin and for A. bidentata (5.40±0.61) was observed in the very same concentration. The highest percentage, maximum number of rootlets/shootlet and mean length of rootlets were observed in 1/2 MS medium supplemented with 1.0 mg/L of IBA. Seventy percentages of plants were successfully established in polycups. Sixty eight percentages of plants were well established in the green house condition. Sixty five percentages of plants were established in the field.</p><p><b>CONCLUSIONS</b>The results have shown that use of nodal buds is an alternative reproducible and dependable method for clonal propagation of A. aspera and A. bidentata. The high rate of direct shoot-root multiplication and their high rate of post-hardening survival indicate that this protocol can be easily adopted for commercial large scale cultivation.</p>

Effect of Huogu II Formula (II) with medicinal guide Radix Achyranthis Bidentatae on bone marrow stem cells directional homing to necrosis area after osteonecrosis of the femoral head in rabbit / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of Huogu II Formula (II) with medicinal guide Radix Achyranthis Bidentatae (Ach) on bone marrow stem cells (BMSCs) homing to necrosis area after osteonecrosis of the femoral head (ONFH) frozen by liquid nitrogen in rabbit as well as to explore the mechanism of prevention and treatment for ONFH.</p><p><b>METHODS</b>The animal model of ONFH was established by liquid nitrogen frozen on the rabbit left hind leg. Forty-eight Japanese White rabbits were randomly assigned to sham-operated group, model group, Huogu II group, and Huogu II plus Ach group, with 12 rabbits in each. During the course of ONFH animal model establishment, all rabbits were subcutaneously injected with recombinant human granulocyte colony-stimulating factor [rhG-CSF, 30 μg/(kg·day) for continuous 7 days]. Meanwhile, normal saline and decoction of the two formulae were administrated by gavage, respectively. White blood cells (WBC) were counted in peripheral blood before and after injection of rhG-CSF. Materials were drawn on the 2nd and 4th weeks after model built; bone glutamine protein (BGP) and bone morphogenetic protein 2 (BMP2) levels in serum were tested. Histopathologic changes were observed by hematoxylin and eosin (HE) staining. BMP2 mRNA levels were detected with in situ hybridization (ISH) staining. 5-Bromo-2'-deoxyuridine (BrdU) and stromal cell derived factor 1 (SDF-1) were measured by immunohistochemical assay in femoral head of the left hind leg.</p><p><b>RESULTS</b>Compared with the shamoperated group, the ratio of empty lacuna, serum BGP, and SDF-1 level in the model group increased significantly, and BMP2 in both serum and femoral head decreased significantly. However, in comparison with the model group, the empty lacuna ratio of Huogu II group and Huogu II plus Ach group decreased obviously in addition to the levels of serum BGP and BMP2, and the expressions of BMP2 mRNA, BrdU, and SDF-1 increased significantly. Above changes were particularly obvious in Huogu II plus Ach group. BGP and SDF-1 on the 2nd week and empty lacuna rate and serum BMP2 level on the 4th week in Huogu II group significantly exceeded their counterparts. On the 2nd week, only in Huogu II plus Ach group that the BrdU counting rose significantly. On the 4th week, empty lacuna rate and serum BMP2 level in Huogu II plus Ach group exceeded those in Huogu II group distinctively.</p><p><b>CONCLUSIONS</b>To a certain extent, the medicinal guide Ach improves the preventive and therapeutic effects of Huogu II Formula on experimental ONFH model. The possible mechanism of this is related to its promoting effect on directional homing of BMSCs to the necrosis area.</p>

Gastroprotective effect of Achyranthes aspera Linn. leaf on rats

OBJECTIVE@#The study was aimed at evaluating the antiulcer activity of ethanolic extract of Achyranthes aspera (EEAA) leaf.@*METHODS@#The anti-ulcer assays were performed on pylorus ligation and chronic ethanol induced ulcer model. The effects of the EEAA on gastric content volume, pH, free acidity, total acidity and ulcer index were evaluated.@*RESULTS@#The percentage of ulcer protection (59.55% and 35.58%) was significantly (P < 0.001) higher in the groups treated with the high dose of EEAA (600 mg/kg), it also reduced the volume of gastric juice and total acidity whereas, gastric pH was increased significantly.@*CONCLUSIONS@#The results of this study show significant gastroprotective activity of EEAA may be due to presence of phyto-constituents like flavanoids, saponins and tannins.

A new feruloyl tyramine glycoside from the roots of Achyranthes bidentata / 中国天然药物

AIM@#To study the chemical constituents of the roots of Achyranthes bidentata Bl.@*METHODS@#The chemical constituents were isolated and purified by macroporous adsorptive resin D101, silica gel, and ODS column chromatographies and preparetive HPLC. Their structures were elucidated on the basis of 1D and 2D NMR analyses.@*RESULTS@#Two feruloyl tyramine glycosides and seven triterpenoid saponins were obtained and identified as N-trans-feruloyl-3-methoxytyramine-4'-O-β-D-glucopyranoside (1), N-trans-feruloyl-3-methoxytyra mine-4-O-β-D-glucopyranoside (2), PJS-1 (3), chikusetsusaponin IVa (4), oleanolic acid 3-O-[β-D-glucuronopy ranoside-6-O-methyl ester]-28-O-β-D-glucopyranoside (5), oleanolic acid 3-O-[β-D-glucuronopyran-oside-6-O-ethylester]-28-O-β-D-glucopyranoside (6), oleanolic acid 3-O-[β-D-glucuronopyranoside-6-O-butyl ester]-28-O-β-D-glucopyranoside (7), ginsenoside R(0) (8) and hederagenin-28-O-β-D-glucopyranosyl ester (9).@*CONCLUSION@#Compound 1 is a new feruloyl tyramine glycoside, while compounds 2 and 9 are reported from A. bidentata for the first time.

Research on relationship between tissue quantitative distribution of 3H-Achyranthes bidentata ecdysterone and channel-tropism of herbal drugs in mice / 中国中药杂志

<p><b>OBJECTIVE</b>To study the relationship between tissue quantitative distribution and pharmacokinetics of 3H-achyranthes bidentata ecdysterone and the channel-tropism of herbal drugs in mice.</p><p><b>METHOD</b>3H-achyranthes bidentata ecdysterone was used as a tracer agent and injected into mice by the caudal vein. In 36 hours, the contents of the tracer agent of samples involving 9 different tracing phases and organ or tissue were determined in order to observe the dynamic quantitative distribution and excretion and pharmacokinetics of 3H-achyranthes bidentata ecdysterone and to understand the channel-tropism of herbal drugs achyranthes bidentata.</p><p><b>RESULT</b>3H-achyranthes bidentata ecdysterone of same organs in different tracing phases and the contents of 3H-achyranthes bidentata ecdysterone in same tracing phases of different organs were significantly different (P<0.01). 3H-achyranthes bidentata ecdysterone was mainly distributed, in the liver, kidney, adrenal gland, small intestine and lung. The concentration-time profiles of achyranthes bidentata ecdysterone in rats injected into mice by the caudal vein were shown to fit a two-compartment open model with half-lives of (778.65 +/- 12.36) min, the elimination of achyranthes bidentata ecdysterone from plasma was found to be in accord with linear kinetics.</p><p><b>CONCLUSION</b>The above mentioned selective distribution of 3H-achyranthes bidentata ecdysterone basically coincides with the meridian affinity and zang fu selection of the traditional Chinese medicine drug Achyranthes bidentata. This study will provide a scientific basis for the channel-tropism of A. bidentata.</p>

Evaluación fotohemolítica in vitro de Cissus sicyoides L y Achyranthes aspera L / In vitro photohemolytic assessment of Cissus sicyoides L y Achyranthes aspera

Los eritrocitos son células útiles para la identificación de agentes potencialmente fototóxicos administrados por vía sistémica, así como para el estudio de los mecanismos de fototoxicidad que involucran procesos de estrés oxidativo. OBJETIVO: evaluar el efecto fotohemolítico de extractos blandos de partes aéreas de Cissus sicyoides L (Vitaceae) y Achyranthes aspera L (Amaranthaceae). MÉTODOS: se utilizó un protocolo in vitro que emplea como modelo biológico eritrocitos humanos, los que se irradian con luz ultravioleta durante 90 min para evaluar el daño en las membranas eritrocitarias, por detección de hemoglobina liberada al medio. RESULTADOS: se observó un leve grado de hemólisis, el efecto fotohemolítico fue inferior a los controles positivos. CONCLUSIONES: los extractos de las plantas se clasificaron como no irritantes, lo cual sugiere que la hemólisis observada puede ser causada por la inestabilidad de la membrana del eritrocito, debido a la presencia de diferentes metabolitos en los extractos estudiados

Erythrocytes are useful cells to identify potentially phototoxic agents provided by systemic administration as well as to study the phototoxicity mechanisms involving oxidative stress processes. OBJECTIVE: to evaluate the photohemolytic effect of soft extracts from aerial parts of Cissus sicyoides L (Vitaceae) and Achyranthes aspera L (Amaranthaceae). METHODS: an in vitro protocol using human erythrocytes as biological model; they were ultraviolet light-radiated for 90 minutes to evaluate damage in erythrocyte membranes on the basis of detected hemoglobin released into the medium. RESULTS: mild hemolysis was observed, being the photohemolytic effect lower than that of positive controls. CONCLUSIONS: the extracts from these plants were rated as non-irritating, which suggests that observed hemolysis may be caused by unstable erythrocyte membranes resulting from the existence of different metabolites in the studied extracts

Evaluación fotohemolítica in vitro de Cissus sicyoides L y Achyranthes aspera L / In vitro photohemolytic assessment of Cissus sicyoides L y Achyranthes aspera

Los eritrocitos son células útiles para la identificación de agentes potencialmente fototóxicos administrados por vía sistémica, así como para el estudio de los mecanismos de fototoxicidad que involucran procesos de estrés oxidativo. OBJETIVO: evaluar el efecto fotohemolítico de extractos blandos de partes aéreas de Cissus sicyoides L (Vitaceae) y Achyranthes aspera L (Amaranthaceae). MÉTODOS: se utilizó un protocolo in vitro que emplea como modelo biológico eritrocitos humanos, los que se irradian con luz ultravioleta durante 90 min para evaluar el daño en las membranas eritrocitarias, por detección de hemoglobina liberada al medio. RESULTADOS: se observó un leve grado de hemólisis, el efecto fotohemolítico fue inferior a los controles positivos. CONCLUSIONES: los extractos de las plantas se clasificaron como no irritantes, lo cual sugiere que la hemólisis observada puede ser causada por la inestabilidad de la membrana del eritrocito, debido a la presencia de diferentes metabolitos en los extractos estudiados(AU)

Erythrocytes are useful cells to identify potentially phototoxic agents provided by systemic administration as well as to study the phototoxicity mechanisms involving oxidative stress processes. OBJECTIVE: to evaluate the photohemolytic effect of soft extracts from aerial parts of Cissus sicyoides L (Vitaceae) and Achyranthes aspera L (Amaranthaceae). METHODS: an in vitro protocol using human erythrocytes as biological model; they were ultraviolet light-radiated for 90 minutes to evaluate damage in erythrocyte membranes on the basis of detected hemoglobin released into the medium. RESULTS: mild hemolysis was observed, being the photohemolytic effect lower than that of positive controls. CONCLUSIONS: the extracts from these plants were rated as non-irritating, which suggests that observed hemolysis may be caused by unstable erythrocyte membranes resulting from the existence of different metabolites in the studied extracts(AU)

Effect of phosphorus on copper tolerance in Achyranthes bidentata / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of phosphorus on copper tolerance in Achyranthes bidentata.</p><p><b>METHOD</b>A PVC pipe experiment was conducted to study the interactive effects of phosphorus (P) and copper (Cu), on growth, elemental accumulation and chemical constituents of A. bidentata. Two levels of elemental P were applied at 0 (P0) and 100 ( P100) mg x kg(-1) soil with 5 levels of Cu at 0 (Cu0), 100 (Cu100), 200 (Cu200), 200 (Cu400), 200 (Cu600) mg x kg(-1) soil, respectively.</p><p><b>RESULT AND CONCLUSION</b>The biomass production between different Cu treatments, phosphorus treatment showed significant differences. The biomass reached the maximum value as the concentration of Cu and P was 100 mg x kg(-1). Low concentration of Cu improved the growth of A. bidentata. The growth was blocked as Cu concentration reached 200 mg x kg(-1) in soil, however the contents of oleanolic acid and ecdysterone in roots of A. bidentata had not influenced by Cu. P could improved the copper tolerance in A. bidentata and increased root yield. The Cu concentration in soil of the cultivation bases must be below 200 mg x kg(-1) in order to produce good quality of medicinal material.</p>

Determination of 5-hydroxymethyl-furfural in root of Achyranthes bidentata expressing different degree of floating sugar / 中国中药杂志

<p><b>OBJECTIVE</b>To study the relation between the content of 5-hydroxymethyl-furfural (5-HMF) and the degree of floating sugar in root of Achyranthes bidentata.</p><p><b>METHOD</b>An HPLC method was applied with a Waters Symmetry C18 3.9 mm x 150 mm (5 microm) column by using methanol-water (12:88) as the mobile phase at a flow rate of 1.0 mL x min(-1) and a UV detection of 280 nm.</p><p><b>RESULT</b>Along with the degree's deepening of floating sugar, the content of 5-HMF varied with the different shades of the sample. The content was 10 times higher in the black sample (highest degree of flowing suger) than that in the yellowish sample (normal). The concentrations of 5-HMT in five yellowish samples of roots of A. bidentata were 0.162 mg x g(-1) to 0.332 mg x g(-1).</p><p><b>CONCLUSION</b>The content increasing of 5-HMF in the root of A. bidentata was related to the degree of flowing sugar.</p>

Study on drug-guide effect of Achyranthes bidentata in Sanmiao pill in arthritic rats / 中国中药杂志

<p><b>OBJECTIVE</b>To study the drug-guiding mechanism of Achyranthes bidentata from Sanmiao pill in arthritic rats.</p><p><b>METHOD</b>The rats were treated by Ferud's complet adjuvant and hot water -bath to establish rat adjuvant arthritis model, then the model rats were divided into three groups, model group, Sanmiao pill groups with A. bidentata. (18 g x kg(-1)) and without A. bidentata. (10 g x kg(-1)), respectively. The heart and foot joints were washed and homogenated to determine the berberine concentration by HPCL in different time after ig and the foot edema was tested by volume method. The pathological changes were observed and hemorheologic parameters were also tested.</p><p><b>RESULT</b>The berberine concentration of foot joint was significantly higher in 2, 4, 6 hour and 14 day in the rats with A. bidentata. The berberine concentration ratio of foot joint and heart was significantly higher in rats with A. bidentata. pharmacodynamic researches showed that A. bidentata could enhance the edema inhibition effect of Sanmiao pill. Hemorheologic researches showed that A. bidentata. could significantly improve the blood viscosity of model rats, the blood high shear viscosity, the blood low shear viscosity and the plasma viscosity were (6.47 +/- 0.57), (9.28 +/- 1.2), (1.94 +/- 0.19) mPa x s respectively.</p><p><b>CONCLUSION</b>A. bidentata. could facilitate the targeted tissue distribution of berberine. The effect was correlative with its blood viscosity improvement.</p>