Subchronic toxicological evaluation of EnXtra™ (standardised extract of Alpinia galanga rhizome) in rats.

OBJECTIVES: Alpinia galanga is a commonly used ingredient in Asian food and traditional medicine. But an extract of the rhizome had never been used commercially in food supplements and functional foods. There is some evidence of safety and tolerability in humans for a proprietary A. galanga rhizome extract (EnXtra™) and it is Generally Recognised as Safe (GRAS) in the US already. However, this botanical ingredient has not been evaluated for its subchronic toxicity in rats to confirm its safety in wider food applications. METHODS: Sprague Dawley rats were orally administered the test item for 90 days by following OECD (Test Guideline: 408), with a recovery period of 28 days. Cumulative effects and No Observed Adverse Effect Level (NOAEL) were estimated. EnXtra™ was administered orally at 0, 1,000, 2,000 and 3,000 mg kg-1 body weight (b. wt.) with additional vehicle and high dose recovery groups. Observations included clinical signs, haematology, clinical chemistry, gross pathology and histopathology. RESULTS: On terminal sacrifice, no treatment-related adverse effects were observed viz., clinical signs, mortality, body weight changes and feed consumption parameters. Haematology, clinical biochemistry and thyroid hormone levels were within the normal range. Further, no treatment-related gross and microscopic pathological lesions were observed across the treatment groups. CONCLUSIONS: Based on the results of the toxicological evaluation, NOAEL of A. galanga rhizome extract (AGRE) was fixed at 3,000 mg kg-1 b. wt. per day and ADI of 1800 mg day-1 in the case of humans.

A 13-week subchronic toxicity study of a Dioscorea Rhizome water extract in rats.

Dioscorea Rhizome is widely used as a traditional herbal medicine to treat asthma, diarrhea, cough, bronchitis, spermatorrhea, leukorrhea, and rheumatoid arthritis. This study investigated the potential subchronic toxicity of a D. Rhizome water extract (DRWE) after repeated oral administration at 0, 800, 2000, and 5000 mg/kg/day in rats for 13 weeks. During the study period, clinical signs, mortality, body weight, food consumption, water consumption, urinalysis, ophthalmoscopy, hematology, serum biochemistry, gross pathology, organ weights, and histopathology were examined. The 13-week repeated oral administration of DRWE to rats resulted in an increased incidence of zona glomerulosa hypertrophy and hyperplasia in the adrenal gland at dose levels of ≥2000 mg/kg/day in both sexes. However, these findings are considered as non-adverse adaptive changes because of minimal histological changes in the lesions, which were not accompanied by any corresponding alterations in serum electrolytes and adrenal gland weight. No treatment-related adverse effects on clinical signs, body weight, food and water consumption, ophthalmic examination, urinalysis, hematology, serum biochemistry, necropsy findings, and organ weights were observed at any dose tested. Under the present experimental conditions, the no-observed-adverse-effect level of the DRWE was considered to be 5000 mg/kg/day in both sexes, and no target organs were identified.

The integrated use of in silico methods for the hepatotoxicity potential of Piper methysticum.

Herbal products as supplements and therapeutic intervention have been used for centuries. However, their toxicities are not completely evaluated and the mechanisms are not clearly understood. Dried rhizome of the plant kava (Piper methysticum) is used for its anxiolytic, and sedative effects. The drug is also known for its hepatotoxicity potential. Major constituents of the plant were identified as kavalactones, alkaloids and chalcones in previous studies. Kava hepatotoxicity mechanism and the constituent that causes the toxicity have been debated for decades. In this paper, we illustrated the use of computational tools for the hepatotoxicity of kava constituents. The proposed mechanisms and major constituents that are most probably responsible for the toxicity have been scrutinized. According to the experimental and prediction results, the kava constituents play a substantial role in hepatotoxicity by some means or other via glutathione depletion, CYP inhibition, reactive metabolite formation, mitochondrial toxicity and cyclooxygenase activity. Some of the constituents, which have not been tested yet, were predicted to involve mitochondrial membrane potential, caspase-3 stimulation, and AhR activity. Since Nrf2 activation could be favorable for prevention of hepatotoxicity, we also suggest that these compounds should undergo testing given that they were predicted not to be activating Nrf2. Among the major constituents, alkaloids appear to be the least studied and the least toxic group in general. The outcomes of the study could help to appreciate the mechanisms and to prioritize the kava constituents for further testing.

Anti-breast cancer and toxicity studies of total secondary saponin from Anemone raddeana Rhizome on MCF-7 cells via ROS generation and PI3K/AKT/mTOR inactivation.

ETHNOPHARMACOLOGICAL RELEVANCE: The rhizome of Anemone raddeana Regel (A. raddeana) is a famous traditional Chinese medicine (TCM) recorded in Chinese Pharmacopoeia for the treatment of carbuncle and swelling. Carbuncle swollen is an explanation of tumor in the theory of TCM and softening and resolving hard mass effects are one of the important pharmacological activities of A. raddeana. AIM OF THE STUDY: We investigated the potential anti-breast cancer effect and toxicological properties of alkali-ethanol extract from A. raddeana, namely total secondary saponin (TSS). MATERIALS AND METHODS: Anti-proliferative effect of total saponin of A. raddeana (ATS) and TSS were tested using MTT assay. Hoechst staining, flow cytometry analysis, DCFH-DA fluorescence microscopy and western blot were carried out to evaluate the mechanisms of action of TSS. The potential anti-breast cancer activity and toxicological properties of TSS were tested in vivo. RESULTS: ATS and TSS could inhibit the proliferation of A549, HepG2, MCF-7, MDA-MB-231 and SKBr-3 cells, especially for MCF-7 cells. Flow cytometry analysis revealed that TSS (10, 12 and 15 µg/ml) could induce cell cycle arrest on G0/G1 phase and promote apoptosis of MCF-7 cells. TSS could increase Bax/Bcl-2 ratio, elevate cytochrome c levels in cytosol and activate caspase-3/9. In addition, TSS also induced ROS generation and inactivated PI3K/AKT/mTOR pathway which may involved in the mitochondrial dysfunction of MCF-7 cells. TSS showed slight toxic at the dosage of 100 and 200 mg/kg by oral administration without any toxic potential for 28 days. TSS (50, 100 and 200 mg/kg) showed significant inhibitory effect on growth of transplanted tumor in mice. At last, twenty-three C-3 monosaccharide oleanane-type triterpene saponins were tentatively identified, which may contributed to the anti-cancer activity of TSS. CONCLUSION: This study demonstrated that TSS exhibited anti-proliferative and pro-apoptosis activities on MCF-7 cells via ROS-mediated activation of mitochondrial apoptosis pathway. TSS might be used as chemotherapeutic agent for the treatment of breast cancer with relatively low toxicity.

Integrative analysis of proteomic and metabonomics data for identification of pathways related to Rhizoma Paridis-induced hepatotoxicity.

Clinical reports on hepatotoxicity that arise from Rhizoma Paridis have recently received widespread attention. Because the hepatotoxicity mechanism is little understood, this research strived to investigate the hepatotoxicity mechanism of Rhizoma Paridis extracts based on iTRAQ quantitative proteomics and metabonomics. The extraction solutions were administrated to rats for 7 days by gavage, and the hepatotoxicity was assessed through quantification of biochemical indexes and Oil red O staining. Additionally, the mechanism of hepatotoxicity was investigated by metabonomics based upon GC-MS and iTRAQ quantitative proteomics. The biochemical and histopathological analysis stood out that Rhizoma Paridis extract could induce liver injury, which was proved by the formation of fat droplets, the changes of mitochondrial structure, and biochemical parameters. The iTRAQ proteomics and metabonomics revealed that Rhizoma Paridis-induced hepatotoxicity was chiefly connected with the abnormal activity of mitochondrion function, which brought about oxidative stress injuries and inflammation, finally causing cell apoptosis. Collectively, we have provided previously uncharacterized hepatotoxic mechanism induced by Rhizoma Paridis and a reference to ensure its safe use in the future.

Comparison of the acute toxicity, analgesic and anti-inflammatory activities and chemical composition changes in Rhizoma anemones Raddeanae caused by vinegar processing.

BACKGROUND: As the dry rhizome of Anemone raddeana Regel, Rhizoma Anemones Raddeanae (RAR), which belongs to Ranunculaceae, is usually used to treat wind and cold symptoms, hand-foot disease and spasms, joint pain and ulcer pain in China. It is well known that the efficacy of RAR can be distinctly enhanced by processing with vinegar due to the reduced toxicity and side effects. However, the entry of vinegar into liver channels can cause a series of problems. In this paper, the differences in the acute toxicity, anti-inflammatory and analgesic effects between RAR and vinegar-processed RAR were compared in detail. The changes in the chemical compositions between RAR and vinegar-processed RAR were investigated, and the mechanism of vinegar processing was also explored. METHODS: Acute toxicity experiments were used to examine the toxicity of vinegar-processed RAR. A series of studies, such as the writhing reaction, ear swelling experiment, complete Freund's adjuvant-induced rat foot swelling experiment and cotton granuloma, in experimental mice was conducted to observe the anti-inflammatory effect of vinegar-processed RAR. The inflammatory cytokines of model rats were determined by enzyme-linked immunosorbent assay (ELISA). Liquid Chromatography-Quadrupole-Time of Flight mass spectrometer Detector (LC-Q-TOF) was used to analyse the chemical compositions of the RARs before and after vinegar processing. RESULTS: Neither obvious changes in mice nor death phenomena were observed as the amount of vinegar-processed RAR in crude drug was set at 2.1 g/kg. Vinegar-processed RAR could significantly prolong the latency, reduce the writhing reaction time to reduce the severity of ear swelling and foot swelling, and remarkably inhibit the secretion of Interleukin-1ß(IL-1ß), Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) proinflammatory cytokines. The content of twelve saponins (e.g., Eleutheroside K) in RAR was decreased after vinegar processing, but six other types (e.g., RDA) were increased. CONCLUSIONS: These results revealed that vinegar processing could not only improve the analgesic and anti-inflammatory effects of RAR but also reduce its own toxicity. TRIAL REGISTRATION: Not applicable.

Leigongteng (Radix et Rhizoma Tripterygii) via compatibility with Jinqiancao (Herba Lysimachiae): its toxicity-reduced efficacy in H22-bearing mice.

OBJECTIVE: To observe toxicity-reduced effects of Leigongteng (Radix et Rhizoma Tripterygii) (LGT) via compatibility with Jinqiancao (Herba Lysimachiae) (JQC) in H22-bearing mice and investigate the possible underlying mechanism, and further explore whether JQC can enhance LGT-evoked anti-tumor effect. METHODS: H22-bearing mice were orally administered with LGT alone and its compatibility with JQC, and tumors, serum, livers and kidneys were collected to evaluate the toxicity-reduced efficacy and the possible mechanism. RESULTS: LGT evoked significantly elevated biochemical indicators including serum alanine / aspartate transaminase (ALT/AST), creatinine (Cr) and urea nitrogen (BUN) as well as pathological damage in mice, which were all obviously reversed by JQC via compatibility at the ratios from 4/1 to 1/4. Further analysis indicated that pro-inflammatory cytokine tumor necrosis factor-alpha (TNF-α), and malondialdehyde (MDA) levels significantly decreased, while anti-inflammatory cytokine interleukin (IL)-10, and glutathione (GSH), GSH-s transferase (GST), GSH peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT) levels all increased in livers and kidneys of mice. Besides, after compatibility with JQC at the ratios of 4/1, 2/1, 1/1, 1/2 and 1/4, LGT-decreased tumor weight was further decreased by 48.4%, 57.3%, 54.0%, 49.3% and 52.9%, respectively (all P < 0.01). CONCLUSION: JQC could reduce LGT-induced hepatotoxicity and nephrotoxicity, and enhance the antitumor efficacy via compatibility with JQC, and the toxicity-reduced mechanism could involve inhibiting hepatic and kidney oxidative stress and inflammation.

Cardiotoxicity evaluation of nine alkaloids from Rhizoma Coptis.

PURPOSE: Alkaloids derived from Rhizoma Coptis (RC) has been widely applied to clinical treatments in China. However, the toxicity of RC and the alkaloids from RC remained controversial. The research is designed to clarify the cardiotoxic compounds found in RC. METHODS: In this study, the real-time cellular analysis cardio system and the high-content analysis were applied to monitor the function of cardiomyocytes (CMs) in the treatment of nine alkaloids in RC. Luciferase-coupled adenosine triphosphate (ATP) assay was used to detect cell viability. RESULTS: The results showed that berberine, palmatine, berbamine, and oxyberberine were cardiotoxic, which resulted in arrhythmia and cardiac arrest on CMs in a time- and dose-dependent manner. Meanwhile, berbamine and oxyberberine caused shrinkage and detachment on CMs at 10 µM. Cytotoxicity was induced by these two compounds with decline in cell index and ATP depletion. Cardiotoxicity or cytotoxicity was not observed in the other five alkaloids within 10 µM. CONCLUSION: For the first time, the cardiotoxicity of the nine alkaloids was evaluated to clarify the cardiotoxic components in RC. Furthermore, the experimental evidences were provided to support the safety of drug application.

Mycotoxins in spices and herbs-An update.

Spices and herbs have been used since ancient times as flavor and aroma enhancers, colorants, preservatives, and traditional medicines. There are more than 30 spices and herbs of global economic and culinary importance. Among the spices, black pepper, capsicums, cumin, cinnamon, nutmeg, ginger, turmeric, saffron, coriander, cloves, dill, mint, thyme, sesame seed, mustard seed, and curry powder are the most popular spices worldwide. In addition to their culinary uses, a number of functional properties of aromatic herbs and spices are also well described in the scientific literature. However, spices and herbs cultivated mainly in tropic and subtropic areas can be exposed to contamination with toxigenic fungi and subsequently mycotoxins. This review provides an overview on the mycotoxin risk in widely consumed spices and aromatic herbs.

[Protection of Herba Scutellariae Barbatae against hepatotoxicity induced by Rhizoma Dioscoreae Bulbiferae and its mechanism].

This study was conducted to test the protective activity of ethanol extract of Herba Scutellariae Barbatae(SE) against hepatotoxicity induced by Rhizoma Dioscoreae Bulbiferae in mice and its mechanism. SE was orally given to mice at various doses, and ethyl acetate fraction of Rhizoma Dioscoreae Bulbiferae(EF, 450 mg·kg(-1)) was also orally given at the same time. After 11 days, serum levels of alanine/aspartate aminotransferase(ALT/AST), alkaline phosphatase(ALP), total protein(TP) and albumin(ALB) were measured, and liver histological examination was conducted. Liver glutathione(GSH) amount, myeloperoxidase(MPO) activity and serum tumor necrosis factor-α(TNF-α), interleukin-6(IL-6) and interferon-γ(IFN-γ) levels were measured. The expression of heme oxygenase-1(HO-1), inhibitor of kappa B(IκB) and nuclear factor κB(NF-κB) p65 were determined by Western blot. The results showed that SE(200 mg·kg-1) reversed EF-induced changes of serum ALT, AST, ALP, TP and ALB. Liver histology also suggests the protection of SE against EF-induced liver injury. SE reduced the increased MPO activity in liver and TNF-α, IL-6, IFN-γ contents in serum, and blocked the decrease in IκB expression and subsequent increase in phosphorylation and nuclear translocation of p65 induced by EF. EF increased liver GSH amount and heme oxygenase-1(HO-1) protein expression in mice. SE increased liver GSH amount, but decreased the expression of HO-1. All those results suggest that SE alleviates liver injury induced by consecutive administration of EF by alleviating inflammatory injury via inhibiting NF-κB signaling pathway and elevating antioxidant capacity.

[Synergistic protective effect of Scutellariae Radix with Phellodendri Chinensis Cortex on Dioscoreae Bulibferae Rhizoma liver toxicity in rats].

Dioscoreae Bulibferae Rhizoma (RDB) is commonly used in clinical Chinese medicine. It has been used in many kinds of traditional Chinese medicine (TCM), but the toxicity of RDB, easily leads to hepatotoxicity. The objective of the present study is to investigate the synergistic protective effect of Scutellariae Radix (SR) with Phellodendri Chinensis Cortex (PCC) on RDB caused liver toxicity in rats. SD female rats were adopted to establish the hepatotoxicity models by RDB (9 g•kg⁻¹, ig) once daily for 28 consecutive days. After 28 days, liver histological changes were observed, and the activity of transaminase and antioxidant enzymes was evaluated. Morphological and biochemical indicators evaluation showed that, Dioscoreae Bulibferae Rhizoma-induced hepatotoxicity models were successful, and the liver cells were dissolved and swelling with fatty degeneration； inflammatory cells were present in gaps; local punctiformed or lamellar hydropic degeneration was found in liver tissues, with partial necrosis. Indexes of liver function (ALT, AST and ALP) were significantly increased (P<0.05 or P<0.01). The combination of SR and PCC has protective effect on RDB-induced hepatotoxicity in rats. SR+PCC exerted the strongest protective effects against RDB-induced hepatotoxicity. SR, PCC, and SB+CP were observed to exhibit hepatoprotective effect as demonstrated by significant decrease in serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) (P<0.05 or P<0.01), and MAD level in liver tissue (P<0.001), significant increase in GSH content in liver tissue (P<0.001), and significant improvement in his to pathologic changes of liver tissues in rats. SR, PCC and their combinations could achieve liver protection effect by reducing ALT, ALP and AST level in serum, increasing GSH level and anti-oxidantability of liver tissues, and reducing hepatic tissue cells injury.

[Toxicokinetics of bakuchiol, hepatic and renal toxicity in rats after single oral administration of Psoraleae Fructus and combination with Glycyrrhizae Radix et Rhizoma].

To study the toxicokinetics of bakuchiol, hepatic and renal toxicity in rats after single oral administration of Psoraleae Fructus and combined with Glycyrrhizae Radix et Rhizoma, in order to provide scientific evidences for clinical safe medication use. A total of 35 SD rats were randomly divided into seven groups: vehicle (distilled water) control group, Glycyrrhizae Radix et Rhizoma group, positive control (aristolochic acid A) group, Psoraleae Fructus (40 g x kg(-1)) group( both male and female rats), Psoraleae Fructus and Glycyrrhizae Radix et Rhizoma (40 +20) g x kg(-1) group (both male and female rats). HPLC-UV method was used to determine the concentration of bakuchiol in rat plasma at different time points after single oral administration. Plasma alanine transaminase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), plasma creatinine (Cr), N-acetyl-ß-D-glucosaminidase (NAG) and kidney injury molecule 1 (Kim-1) were measured after administration for 24 h. The main toxicokinetics parameters of bakuchiol in rats exert significantly gender difference. When Psoraleae Fructus combination with Glycyrrhizae Radix et Rhizoma, the total area under the plasma concentration-time curve( AUC), C(max), and plasma clearance (CL) of bakuchiol were increased, respectively; CL, half-life (t½) were decreased, and T(max) were prolonged. The biochemical indicators (including ALT, AST, BUN, Cr and KIM-1 level) in different dose of Psoraleae Fructus groups, were found no statistically significant difference when compared with vehicle control group. The level of NAG in both Psoraleae Fructus and compatibility with Glycyrrhizae Radix et Rhizoma groups were significant increased (P < 0.05). There are obvious effects on toxicokinetics of bakuchiol in rats when Psoraleae Fructus combined with Glycyrrhizae Radix et Rhizoma. Renal toxicity induced by Psoraleae Fructus at high dose was observed after single oral administration and no liver damage in rats was found.

An experimental study on providing a scientific evidence for seven-time alcohol-steaming of Rhei Rhizoma when clinically used.

BACKGROUND: Rhei Rhizoma (RR) has been widely used as laxative and processed to alter its therapeutic actions or reduce its side effects. In this study, we evaluated experimentally the clinical application guideline that RR should be alcohol-steamed seven times before being used in elderly patients, as described in Dongeuibogam, the most famous book on Korean traditional medicine. METHODS: Unprocessed RR (RR-U) was soaked in rice wine, steamed and then fully dried (RR-P1). The process was repeated four (RR-P4) or seven times (RR-P7). Reversed-phase high-performance liquid chromatography was used to determine the RR-U, RR-P1, RR-P4 and RR-P7 (RRs) constituents. To evaluate the effect of RRs on liver toxicity, human hepatoma cells (HepG2) were treated with RRs at 100 µg/mL for 4 h and then cell viabilities were measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. To confirm the effects in vivo, 5-week-old male Sprague-Dawley rats were treated with RRs at 3 g/kg/day for 21 days. Body weight and serum biochemical parameters were measured and liver histology was assessed. RESULTS: The levels of sennosides decreased in processed RRs in an iteration-dependent manner, while the emodin level was unaffected. In HepG2 cells, cell viability was reduced with RR-U, while the toxicity decreased according to the number of processing cycles. The changes in body weight, relative liver weight and liver enzymes of RR-U-treated rats were reduced in processed RRs-treated rats. Histopathological analysis indicated swelling and cholestasis improved following seven times alcohol-steaming cycles. CONCLUSIONS: These results provide experimental evidence that RR-P7 almost completely reduces RR hepatotoxicity.

Acute toxicity and long-term safety evaluation of the crude extract from rhizomes of Limonium brasiliense in mice and rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Limonium brasiliense (Boiss.) Kuntze, Plumbaginaceae, popularly known as baicuru, has been used in folk medicine to treat menstrual cramps and to regulate menstrual periods. However, little is known about its safety. This study evaluated the safety through in vivo tests of the acute, long-term, and liver toxicity, and the mutagenicity of the crude extract (CE) from rhizomes of L. brasiliense. MATERIALS AND METHODS: The acute toxicity was assessed in Swiss mice, and the chronic toxicity in Wistar rats. Male and female mice received the CE orally in single doses of 1.0, 2.0, 3.0, 4.0, or 5.0 g/kg. Clinical changes and mortality rate were used as parameters to assess the acute toxicity. In the long-term evaluation, male and female Wistar rats were treated orally with daily doses of the CE (50, 100, or 200 mg/kg) for 90 days. Assessments of weight, behavior and food intake, urinalysis, biochemical and hematological analyses, as well as macro- and microscopic observations of several organs were performed. The redox state of the liver was evaluated as a means of investigating the liver toxicity, and the micronucleus test to assess mutagenicity was also performed. RESULTS: Evaluation of acute toxicity indicated no apparent clinical change in the animals; the LD50 was 4.8 g/kg. Evaluation after 90 days administration showed that the CE, even in higher doses than are considered therapeutic, appeared to be safe. The micronucleus test demonstrated a low mutagenic potential for the CE. CONCLUSION: Our results showed that treatment with the CE from L. brasiliense caused low or no toxicity, as assessed using these doses and evaluation methods.

[Study on intersection and regulation mechanism of "efficacy-toxicity network" of aconite in combination environment of Sini decoction].

OBJECTIVE: To explore the intersection and regulation mechanism of "efficacy-toxicity network" of Glycyrrhizae Radix et Rhizoma, Zingiberis Rhizoma and Aconiti Lateralis Radix Praeparata's action gene in the combination environment of Sini decoction with the network pharmacological method. METHOD: The gene interaction network of Aconiti Lateralis Radix Praeparata, Glycyrrhizae Radix et Rhizoma, Zingiberis Rhizoma were mined and established with Cytoscape software and Agilent literature search plug-in. The "efficiency-toxicity network" intersection of Aconiti Lateralis Radix Praeparata was formed according to its effects in anti-heart failure, neurotoxicity and cardiotoxicity. The target genes were clustered with Clusterviz plug-in. And the possible pathways of the "efficacy-tox- icity network" intersection of Glycyrrhizae Radix et Rhizoma, Zingiberis Rhizoma and Aconiti Lateralis Radix Praeparata were forecasted in DAVID database. RESULT: There were five genes related to neurotoxicity, cardiotoxicity and anti-heart failure function of Aconiti Lateralis Radix Praeparata, namely AKT1, BAX, HCC, IL6 and IL8, which formed 47 nodes genes in the "efficiency-toxicity network" intersection of Aconiti Lateralis Radix Praeparata. There were 29 and 27 coincident genes in the "efficiency-toxicity network" of Glycyrrhizae Radix et Rhizoma, Zingiberis Rhizoma and Aconiti Lateralis Radix Praeparata. There were 23 and 17 possible regulatory pathways. CONCLUSION: In the combination environment of Sini decoction, Glycyrrhizae Radix et Rhizoma and Zingiberis Rhizoma may regulate the efficiency-toxicity network of Aconiti Lateralis Radix Praeparata by influencing immune-inflammatory signaling pathway, apoptosis-autophagy signaling pathway, nerve cell and myocardial ischemia and hypoxia protection signaling pathways.

[Investigating mechanism of toxicity reduction by combination of Glycyrrhizae Radix et Rhizoma and Aconiti Lateralis Radix Preparata on terms of proteins self-assembly].

The combination of Glycyrrhizae Radix et Rhizoma and Aconiti Lateralis Radix Preparata can increase efficacy and decrease toxicity. This study started from the phenomena of protein self-assembly in the mixed decoction of Glycyrrhizae Radix et Rhizoma with Aconiti Lateralis Radix Preparata. The attenuated mechanism was explored between the combination of Glycyrrhizae Radix et Rhizoma and Aconiti Lateralis Radix Preparata by using the protein of Glycyrrhizae Radix et Rhizoma and aconitine which was the major toxic component of Aconiti Lateralis Radix Preparata. Glycyrrhizae Radix et Rhizoma protein with aconitine could form stable particles which particle mean diameter was (206.2 ± 2.02) nm and (238.20 ± 1.23) nm at pH 5.0 in normal temperature. Through the mouse acute toxicity experiment found that injection of aconitine monomer all mice were killed, and injection of Glycyrrhizae Radix et Rhizoma protein-aconitine particles with the same content of aconitine all mice survived. Survey the stability of Glycyrrhizae Radix et Rhizoma protein-aconitine shows that the colloid particles is stable at room temperature, and it has the possibility to candidate drug carrier. Glycyrrhizae Radix et Rhizoma protein can reduce the toxicity of aconitine through self-assembly.

Effect of roucongrong (Herba Cistanches Deserticolae) on reproductive toxicity in mice induced by glycoside of Leigongteng (Radix et Rhizoma Tripterygii).

OBJECTIVE: The purpose of this research is to study the effect of Roucongrong (Herba Cistanches Deserticolae) on reproductive toxicity in mice induced by a glycoside extracted from Leigongteng (Radix et Rhizoma Tripterygii) (GRT). METHODS: Forty-eight BALB/c mice were randomly divided into two groups in the ratio of 1:3, 12 in one group and 36 in the other. The 12-mouse group was the control group that was intragastrically administered physiological saline for 3 weeks. The 36 mice in the other group were given 30 mg x kg(-1) x d(-1) GRT for 3 weeks, then randomly divided into 3 subgroups: the model group, GRT group and Roucongrong (Herba Cistanches Deserticolae) group, with 12 mice in each group. In the model group, 0.25 mL physiological saline was intragastrically administered; in the GRT group, GRT, 0.25 mL at 30 mg x kg(-1) x d(-1) was intragastrically administered once a day; in the Roucongrong (Herba Cistanches Deserticolae) group, mice were administered Roucongrong (Herba Cistanches Deserticolae) decoction equivalent to 0.25 mL at a final dose of 10 g x kg(-1) x d(-1) crude drug (calculated as per 20 times of 0.5 g x kg(-1) x d(-1) for adults), and GRT 0.25 mL at 30 mg x kg(-1) x d(-1) daily. After another 3 weeks of exposure, expression levels of the reproduction-related genes DEAD (Asp-Glu-Ala-Asp) box polypeptide 3, Y-linked, B-cell CLL/lymphoma 6 and Signal transducer and activator of transcription 3 were evaluated. RESULTS: After 6 weeks of GRT treatment, the spermatogenic cell population in the convoluted tubule of testis was in disorder and the tubule cavity expanded. Sertoli cell and Leydig cells exhibited atrophy or disappeared. The number of sperm decreased. The spermatogenic cell level of testis for male mice was ranked in order and sperm was produced in the cavity of the spermatogenic cell. The expression levels of DDX3Y, BCL6 and STAT3 were CONCLUSION: GRT affected reproduction-related genes. Roucongrong (Herba Cistanches Deserticolae) reversed reproductive toxicity in mice induced by GRT.

[Study on new toxicity-reducing methods of pinellia rhizoma prepared by ethanol (I)-new methods and technology].

OBJECTIVE: To explore new toxicity-reducing methods of Pinellia Rhizoma prepared by ethanol and the latest technical parameters. METHOD: Pinellia Rhizoma is prepared with ethanol. The orthogonal experimental design was adopted for investigating amount of ethanol, preparing time, ethanol concentration and preparing temperature. The optimal technology was determined by the comprehensive score of toxicological indicators of PGE2 content of rat celiac percolate, with the rabbit conjunctival irritation test as the intuitive validation on toxicology reduction. The pharmacodynamics validation was used to determine the reasonability of the preparation process. RESULT: The optimal technology was that Pinellia Rhizoma was prepared by 75% ethanol at the temperature of 60 degrees C by 4 days, and then dried. The effect of relieving cough, reducing sputum and anti-inflammatory of Pinellia Rhizoma is not reduced after prepared by ethanol. CONCLUSION: The optimal technology of Pinellia Rhizoma prepared by ethanol is simple and reasonable that it can be used as the new method to reduce toxicity and keep efficacy of Pinellia Rhizoma.

The furano norclerodane diterpenoid disobulbin-D induces apoptosis in normal human liver L-02 cells.

Disobulbin-D (DBD), a hepatotoxic furano norclerodane diterpenoid, was isolated by bio-guided fractionation from the rhizome of Dioscorea bulbifera L. In working toward elucidating the cellular and molecular mechanisms of DBD toxicity, we treated normal human liver cell line L-02 cells with DBD in vitro and evaluated its toxicity in terms of cell viability, morphologic changes, induction of apoptosis/necrosis, and caspase 3 activity. The viability of L-02 cells was inhibited by DBD in a concentration and time-dependent manner. Apoptosis was supported by the Annexin V and propidium iodide assay, Hoechst 33258 staining, and the occurrence of a sub-G(1) peak. DBD can cause an increase in caspase 3 activity, and pretreatment with Ac-DEVD-CHO blocked cell death and attenuated the apoptosis, showing that DBD-induced L-02 cell apoptosis is caspase 3-dependent. These results suggest that the effects of DBD on the growth of normal human liver L-02 cells may be due to its induction of cell apoptosis, which may also explain the toxicity observed in the plants containing furano clerodane diterpenoids.

Kava hepatotoxicity in traditional and modern use: the presumed Pacific kava paradox hypothesis revisited.

Kava, a Pacific herb consumed worldwide for medicinal, recreational and cultural purposes, has been associated with rare hepatotoxicity, and there is currently a critical need to determine this causation. The previously proposed Pacific kava paradox was based on the theory that kava hepatotoxicity was not observed following use of traditional aqueous extracts in the Pacific region, but was restricted to use of Western acetonic and ethanolic extracts. Subsequent cases analyzed by the World Health Organization and published case reports revealed that traditional aqueous extracts used in New Caledonia, Australia, the USA and Germany may also be hepatotoxic; thus, there is no longer a basis to sustain the previously proposed Pacific kava paradox. It appears that the primary cause of toxicity may reside in the time before the preparation of the various kava extracts, possibly attributed to poor quality of the raw material caused by mould hepatotoxins. Rigorous testing of kava raw material is urgently advised, in addition to Pan-Pacific kava manufacturing quality standards.