Anemarrhena Research Articles

Herbal Medicine and False-Positive Results on Lymphocyte Transformation Test

In vitro mitogenic activity of 16 herbs and 3 Kampo (herbal medicine) formulae have been reported in experimental studies. It is not known how many herbs and Kampo formulae in total have mitogenic activity. Lymphocyte transformation test (LTT) is generally utilized to diagnose drug-induced liver injury. In LTT, mitogenic activity is assessed by measuring 3H-thymidine incorporation. The objective of the present study was to determine which herbs and which Kampo formulae caused false-positivity on LTT. We examined 2496 summaries of all admission records from 1979 to 1999 in our department. We selected patients in whom liver injuries were diagnosed as definitely unrelated to Kampo medication. In these patients, LTT was performed for some herbs contained in the suspect Kampo medicines, resulting in positive LTT for 17 herbs: Evodiae Fructus (Goshuyu), Zizyphi Fructus (Taiso), Ginseng Radix (Ninjin), Zingiberis Rhizoma (Shokyo), Hoelen (Bukuryo), Aconiti Tuber (Bushi), Angelicae Radix (Toki), Cnidii Rhizoma (Senkyu), Rehmanniae Radix (Jio), Ephedrae Herba (Mao), Anemarrhenae Rhizoma (Chimo), Cinnamomi Cortex (Keihi), Bupleuri Radix (Saiko), Artemisiae Capillari Spica (Inchinko), Persicae Semen (Tonin), Moutan Cortex (Botanpi) and Paeoniae Radix (Shakuyaku). These results were considered false-positive, because the results were observed in the "definitely unrelated" patients. Mitogenic activity inherent to some herbs and Kampo formulae may sometimes cause false-positivity on LTT in clinical situations. These examples suggest that LTT for Kampo formulae may be unreliable as a diagnostic method for drug-induced liver injury.

Effect of timosaponin A-III, from Anemarrhenae asphodeloides Bunge (Liliaceae), on calcium mobilization in vascular endothelial and smooth muscle cells and on vascular tension

The effects of timosaponin A-III (TA-III), from Rhizoma Anemarrhenae, on Ca 2+ mobilization in vascular endothelial cells and smooth muscle cells and on vascular tension have been explored. TA-III increased intracellular Ca 2+ concentrations ([Ca 2+] i) in endothelials cells at a concentration larger than 5 μM with an EC 50 of 15 μM, and increased [Ca 2+] i in smooth muscle cells at a concentration larger than 1 μM with an EC 50 of 8 μM. Within 5 min, the [Ca 2+] i signal was composed of a gradual rise, and the speed of rising depended on the concentration of TA-III. The [Ca 2+] i signal was abolished by removing extracellular Ca 2+ and was recovered after reintroduction of Ca 2+. The TA-III-induced [Ca 2+] i increases in smooth muscle cells were partly inhibited by 10 μM nifedipine or 50 μM La 3+, but was insensitive to 10 μM verapamil and diltiazem. TA-III (10–100 μM) inhibited 0.3 μM phenylephrine-induced vascular contraction, which was abolished by pretreatment with 100 μM N ω-nitro-L-arginine (L-NNA) or by denuding the aorta. TA-III also increased [Ca 2+] i in renal tubular cells with an EC 50 of 8 μM. Collectively, the results show for the first time that TA-III causes [Ca 2+] i increases in the vascular system. TA-III acted by causing Ca 2+ influx without releasing intracellular Ca 2+. TA-III induced relaxation of phenylephrine-induced vascular contraction via inducing release of nitric oxide from endothelial cells.

Growth inhibition and apoptosis of gastric cancer cell lines by Anemarrhena asphodeloides Bunge.

In this study, we aimed to determine the growth inhibition and the induction of apoptotic cell death brought about by the herb Anemarrhena asphodeloides Bunge in gastric cancer cell lines, and to clarify the mechanism of this apoptosis. Water-soluble ingredients of A. asphodeloides, and the gastric cancer cell lines, MKN45 and KATO-III, were used in vitro. Growth inhibition, induction of cell death, morphological features, the presence of DNA ladders, increases in caspase-3-like activity, the effects of a caspase-3 inhibitor on apoptotic cell death, and the release of cytochrome c by A. asphodeloides were analyzed. A. asphodeloides inhibited the growth and decreased the viability of the gastric cancer cell lines. The viability of normal skin fibroblasts in the presence of low concentrations of A. asphodeloides was higher than that of gastric cancer cells. Apoptotic bodies and DNA ladders were observed to be induced in MKN45 and KATO-III by A. asphodeloides. The caspase 3 inhibitor, Ac-DEVD-CHO, inhibited the apoptotic cell death of gastric cancer cells induced by A. asphodeloides. The caspase 3-like activity in MKN45 and KATO-III cells increased after the addition of A. asphodeloides. Cytochrome c was released from mitochondria into the cytosol 8 h after the addition of A. asphodeloides, and reached a peak at 16 h. The peak of cytochrome c release was earlier than that of caspase 3-like activity. We concluded that A. asphodeloides inhibited the growth of the gastric cancer cell lines MKN45 and KATO-III and induced apoptosis. The apoptosis of MKN45 and KATO-III cells induced by A. asphodeloides was associated with the release of cytochrome c from the mitochondria, followed by an increase in caspase 3-like activity.

Testosterone 5alpha-reductase inhibitory active constituents from Anemarrhenae Rhizoma.

The diethyl ether extract of Anemarrhenae Rhizoma (rhizomes of Anemarrhena asphodeloides Bunge) showed testosterone 5alpha-reductase inhibitory activity. Two major constituents, cis-hinokiresinol (1) and 2,6,4'-trihydroxy-4-methoxybenzophenone (2) were identified as the active principles. The inhibitory activity of 1 was superior to that of ethinylestradiol, but that of 2 was weak.

Antidiabetic activity of a xanthone compound, mangiferin.

Mangiferin (MF) isolated from Anemarrhena asphodeloides Bunge rhizome, was tested for antidiabetic activity in KK-Ay mice, an animal model of type-2 diabetes. MF lowered the blood glucose level of KK-Ay mice 3 weeks after oral administration (p < 0.01). However, no effect on the blood glucose level in normal mice was seen, indicating that MF could be useful in treating type-2 diabetes. In addition, MF improved hyperinsulinemia and, on insulin tolerance test, reduced blood glucose levels of KK-Ay mice. From these findings, it seems likely that MF exerts its antidiabetic activity by decreasing insulin resistance.

Antidiabetic activity of the rhizoma of Anemarrhena asphodeloides and active components, mangiferin and its glucoside.

The antidiabetic activity of the rhizoma of Anemarrhena asphodeloides was investigated in KK-Ay mice, an animal model of genetic type 2 diabetes. The water extract of the rhizoma (AA) (90 mg/kg) reduced blood glucose levels from 570 +/- 29 to 401 +/- 59 mg/dl 7 h after oral administration (p<0.05) and also tended to reduce serum insulin levels in KK-Ay mice. AA-treated KK-Ay mice had significantly reduced blood glucose levels in an insulin tolerance test. Based on these results, the antidiabetic mechanism of AA may be due to decreased insulin resistance. In addition, the active components of AA were confirmed to be mangiferin and its glucoside.

In vitro synergism between nyasol, an active compound isolated from Anemarrhena asphodeloides, and azole agents against Candida albicans.

The antifungal activity of nyasol (NYS) alone or with various antifungal agents was measured in vitro against Candida albicans, Aspergillus fumigatus, and Trichophyton mentagrophytes. NYS is a compound recently purified from a medicinal plant, Anemarrhena asphodeloides. Among 12 agents, miconazole (MCZ), ketoconazole (KCZ), clotrimazole (CTZ), and cerulenin showed marked synergistic effects against C. albicans. The fractional inhibition concentration (FIC) indices against 4 strains of C. albicans were 0.067-0.31 for MCZ plus NYS, 0.078-0.31 for KCZ plus NYS, and 0.098-0.13 for CTZ plus NYS. These values indicate the possibility of using NYS as an adjuvant to azole agents in the chemotherapy of candidiasis.

Effect of six steroidal saponins isolated from anemarrhenae rhizoma on platelet aggregation and hemolysis in human blood

Six steroidal saponins were isolated from Anemarrhena asphodeloides Bunge (Liliaceae), a traditional chinese medicine, and named anemarrhenasaponin I (An-I), anemarrhenasaponin Ia (An-Ia), timosaponin B-I (TB-I), timosaponin B-II (TB-II), timosaponin B-III (TB-III), and timosaponin A-III (TA-III). The effects of these six compounds on platelet aggregation and hemolysis in human blood were studied. All these compounds provoked remarkable inhibiting effect on platelet aggregation, and activated partial thromboplastin times (APTT) are sensitive to the presence of these six compounds. Using an in vitro system, APTT was delayed with the increment of the concentrations of these six compounds. In these six compounds, only timosaponin A-III appeared a strong effect on hemolysis, and anemarrhenasaponin Ia had a slight effect on hemolysis, other had no effect on hemolysis. These results suggested that these steroidal saponins isolated from Anemarrhena asphodeloides Bunge (Liliaceae) might be used as a novel antithrombotic therapeutic agents in post-myocardial infarction.

Effect of Steroidal Saponins of Anemarrhenae Rhizoma on Superoxide Generation in Human Neutrophils

Effect of six steroidal saponins isolated from Anemarrhenae rhizoma on superoxide generation in human neutrophils was investigated. The steroidal saponins examined were anemarrhenasaponin-I (An-I), anemarrhenasaponin-Ia (An-Ia), timosaponin B-I (TB-I), timosaponin B-II (TB-II), timosaponin B-III (TB-III) and timosaponin A-III (TA-III). An-I, An-Ia, and TB-III suppressed the superoxide generations induced by N-formyl-methionyl-leucyl-phenylalanine (fMLP) and arachidonic acid (AA) in a concentration-dependent manner, but enhanced that induced by phorbol 12-myristate 13-acetate (PMA). While TB-II also suppressed and enhanced the superoxide generations induced by fMLP and PMA, respectively, the compound significantly enhanced the AA-induced superoxide generation. TB-I enhanced the fMLP-induced superoxide generation in a low concentration range (peak at 40 μM), gave no effect on the PMA-induced superoxide generation and weakly enhanced the AA-induced superoxide generation. TA-III enhanced the fMLP-induced superoxide generation more than twice as much as that by TB-I in the same concentration range. However, TA-III enhanced the PMA-induced superoxide generation and most significantly suppressed the AA-induced superoxide generation.

Norlignans with Hyaluronidase Inhibitory Activity fromAnemarrhena asphodeloides

Assay-guided fractionation of an MeOH extract of ANEMARRHENA ASPHODELOIDES Bunge has furnished two hyaluronidase inhibitory norlignans, CIS-hinokiresinol ( 1) and 1,3-di- P-hydroxyphenyl-4-penten-1-one ( 2), and an inactive norlignan 4'-methyl- CIS-hinokiresinol ( 3). Compound 2 is a new compound.

Detection of antifungal activity in Anemarrhena asphodeloides by sensitive BCT method and isolation of its active compound.

Antifungal activity was detected from Anemarrhena asphodeloides by the Bio-Cell Tracer (BCT) method. An active fraction was separated by silica gel column chromatography and reverse-phase HPLC. The molecular weight was determined by GC-MS, and the molecular structure was analyzed by IR, (1)H NMR, and (13)C NMR. The isolated compound was found to be identical to nyasol, (Z)-1, 3-bis(4-hydroxyphenyl)-1,4-pentadiene, which formerly appeared in the literature without any remark on the antifungal activity. This compound showed antimicrobial activity against 38 strains of fungi and five strains of bacteria. The minimum inhibitory concentration (MIC) ranged from 12.5 to 200 microg mL(-)(1), except for two strains based on the broth dilution method.

New antidiabetic compounds, mangiferin and its glucoside.

Mangiferin (MF) and its glucosides (mangiferin-7-O-beta-glucoside) (MG) isolated from Anemarrhena asphodeloides Bunge rhizome, were tested for their antidiabetic activity in KK-Ay mice, an animal model of non-insulin-dependent diabetes mellitus (NIDDM). MF and MG lowered the blood glucose level of KK-Ay mice after oral administration. However, no affect on the blood glucose level in normal mice was seen, indicating that MF and MG are useful in treating NIDDM. In addition, MF or MG improved hyperinsulinemia in KK-Ay mice. From these findings, it seems likely that MF and MG exert their its antidiabetic activity by increasing insulin sensitivity.

Hypoglycaemic Effects ofShokatsu-Cha(Xiao-Ke-Ca) in Streptozotocin-induced Diabetes Mellitus

Shokatsu-cha is a novel formula for diabetic mellitus based on the traditional Chinese medicine and composed of Dioscoreae rhizoma, Adenophorae radix, Rehmanniae radix, Anemarrhenae rhizoma, Platycodi radix, Salviae miltiarrhizae radix, Epimedii folium and Lycii fructus. The hypoglycaemic effect of Shokatsu-cha was evaluated in streptozotocin (STZ)-induced diabetic mice and rats. When orally administered once a day for 10 days to the mice intraperitoneally injected with STZ, the Shokatsu-cha extract suppressed the increase of the blood glucose level during administration. In the in situ intestine circulation method, the Shokatsu-cha extract and its ethanol-eluted fraction inhibited tbe glucose absorption in STZ-induced diabetic rats. The ethanol-eluted fraction also inhibited the increase of blood glucose level in an oral glucose tolerance test using diabetic mice. This study indicates that a part of the hypoglycaemic effect of Shokatsu-cha is based on its inhibitory action on intestinal glucose absorption in diabetes.

Combined potentiating effect of byakko-ka-ninjin-to, its constituents, rhizomes of Anemarrhena asphodeloides, tomosaponin A-III, and calcium on pilocarpine-induced saliva secretion in streptozocin-diabetic mice.

The effects on pilocarpine-induced saliva secretion by a hot aqueous extract of Byakko-ka-ninjin-to (BN), its constituents, rhizomes of Anemarrhena asphodeloides, three saponins (pseudoproto-timosaponin-AIII (An-S-1), proto-timosaponin-AIII (An-S-2) and timosaponin-AIII (An-S-3)) and calcium were examined in streptozocin (STZ)-induced diabetic and normal mice. The hot aqueous extracts of BN (250 and 500 mg/kg, i.p.) and Anemarrhena (170 and 340) mg/kg, i.p.) significantly promoted salivary flow in the diabetic animals, but suppressed it in the normal controls. An-S-2 and An-S-3 but not An-S-1 (10 mg/kg, i.p.), significantly promoted salivary How in the diabetic animals. The potency order was An-S-3 >> An-S-2 >> extract. The hot aqueous extracts of BN and Anemarrhena increased the protein content of saliva in a dose-dependent manner. Combination of An-S-3 (0.1 mg/kg, i.p.) with CaCl2 (2 and 4 mg/kg, i.p.) potentiated salivary flow compared with the respective effect of each on its own. These results demonstrated that 1) An-S-3 was mainly responsible for saliva secretion of the hot aqueous extract, and 2) the effect of An-S-3 was potentiated by combination with calcium, suggesting combined effects of Byakko-ka-ninjin-to containing Anemarrhena asphodeloides and gypsum fiber (calcium).

Isolation of Pseudoprototimosaponin AIII from Rhizomes of Anemarrhena asphodeloides and Its Hypoglycemic Activity in Streptozotocin-Induced Diabetic Mice

A hot-H2O extract of rhizomes of Anemarrhena asphodeloides, the Japanese sino-medicine "chimo," lowered the blood glucose level in alloxan-diabetic mice. Hypoglycemic activity-guided fractionation isolated a new glycoside, pseudoprototimosaponin AIII [1], which was compared with chemically known prototimosaponin AIII [2]. These compounds exhibited hypoglycemic effects in a dose-dependent manner in streptozotocin-diabetic mice but showed no effects on glucose uptake and insulin release, suggesting that the hypoglycemic mechanism may be due to inhibition of hepatic gluconeogenesis and/or glycogenolysis.

知母のサポニン成分について

Three steroidal saponins 3, 4 and 5a were newly isolated from Anemarrhenae Rhizoma. Compounds 3 and 4 were identical with desgalactotigonin and F-gitonin, respectively. Compound 5a was established as (25S)-26-O-beta-D-glucopyranosyl-22-hydroxy-5 beta-furostane-3 beta,26-diol 3-O-beta-D-glucopyranosyl-(1----2)-O-beta-D-galactopyranoside on the basis of chemical and spectroscopic evidence.

放射線障害防護薬剤に関する研究(第33報)放射線皮膚障害に対する各種化合物の防護機構

The radiation protective mechanisms on skin injury induced by soft X-irradiation were investigated by use of various radiation protective agents such as sulfur compounds (MEA, MEG, thiourea), nucleic acid constitutional compounds (adenosine, inosine), antioxidative compounds (sesamol, ferulic acid, ascorbic acid), crude drugs (Rosae Fructus, Anemarrhenae Rhizoma, Trapae Fructus, Forsythiae Fructus, Aloe arborescens). Scavenge action of activated oxygen, inhibitory effect of lipid peroxidation, induction of antioxidative protein and protective effect against damage of deoxyribonucleic acid and superoxide dismutase by X-irradiation were evaluated as the radiation protective mechanisms, and relationship between these results and protective effect of skin injury induced by radiation was studied.

Studies on chemical protectors against radiation. XXVI. Protective effect of various extracts on crude drugs on skin injury induced by X-irradiation

The protective potency against skin injury on mice induced by X-irradiation was studied by use of 72 extracts of crude drugs. The protective potency was determined according to the degrees on skin injury after irradiation of 1100R, 30 k Vp soft X-ray. As a result of these study, 16 kinds of crude drugs such as Rosae Fructus, Aloe arborescens (Herba), Citri Leiocarpae Exocarpium, Schizonepetae Spica, Evodiae Fructus, Bupleuri Radix, Corni Fructus, Perillae Herba, Anemarrhenae Rhizoma, Menthae Herba, Trapae Fructus, Angelicae Dahuricae Radix, Sinomeni Caulis et Rhizoma, Ephedrae Herba, Acer nikoense (Cortex), Forsythiae Fructus, revealed protective potencies on skin injury.

Zhi-mu saponin inhibits α-fetoprotein gene expression in developing rat liver

1. 1. A saponin isolated from the Chinese herb zhi-mu ( Anemarrhena asphodeloides Bunge) modifies α-fetoprotein production when injected into newborn rats. 2. 2. The serum level of AFP was determined quantitatively by immunorocket electrophoresis. 3. 3. AFP serum levels were reduced to 60% of the control by zhi-mu saponin (ZMS). 4. 4. The lower AFP level in drug treated rat serum is not due to a change in the pattern of serum AFP variants. 5. 5. AFP mRNA levels in ZMS-treated rat livers, measured by RNA dot hybridization, decreased to about 50% of control levels after 4 days treatment. 6. 6. Results from tritium labeled dexamethasone competition assays suggest that ZMS may act on AFP gene expression through glucocorticoid receptor mediated action.

Screening for molluscicidal activity in crude drugs.

Thirty-four extracts of crude drugs and medicinal plants have been screened for activity against Oncomelania nosophora, the intermediate host of the Japanese strain of Schistosoma japonicum. Strong molluscicidal activity was found in the MeOH extract of Anemarrhenae Rhizoma. Although timosaponin A-III, one of the main saponins of Anemarrhenae Rhizoma, showed very strong killing activity, markogenin-3-O-beta-D-glucopyranosyl (1-2)-O-beta-D-galactopyranoside having the same glycosidic linkage was found to be inactive.