ABSTRACT
Based on the sarcoma receptor coactivator(Src)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt) signaling pathway, the mechanism of action of bulleyaconitine A in the treatment of bone destruction of experimental rheumatoid arthritis(RA) was explored. Firstly, key targets of RA bone destruction were collected through GeneCards, PharmGKB, and OMIM databa-ses. Potential targets of bulleyaconitine A were collected using SwissTargetPrediction and PharmMapper databases. Next, intersection targets were obtained by the Venny 2.1.0 platform. Protein-protein interaction(PPI) network and topology analysis were managed by utilizing the STRING database and Cytoscape 3.8.0. Then, Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses were conducted in the DAVID database. AutoDock Vina was applied to predict the molecular docking and binding ability of bulleyaconitine A with key targets. Finally, a receptor activator of nuclear factor-κB(RANKL)-induced osteoclast differentiation model was established in vitro. Quantitative real-time polymerase chain reaction(qRT-PCR) was used to detect the mRNA expression levels of related targets, and immunofluorescence and Western blot were adopted to detect the protein expression level of key targets. It displayed that there was a total of 29 drug-disease targets, and Src was the core target of bulleyaconitine A in anti-RA bone destruction. Furthermore, KEGG enrichment analysis revealed that bulleyaconitine A may exert an anti-RA bone destruction effect by regulating the Src/PI3K/Akt signaling pathway. The molecular docking results showed that bulleyaconitine A had better bin-ding ability with Src, phosphatidylinositol-4,5-diphosphate 3-kinase(PIK3CA), and Akt1. The result of the experiment indicated that bulleyaconitine A not only dose-dependently inhibited the mRNA expression levels of osteoclast differentiation-related genes cathepsin K(CTSK) and matrix metalloproteinase-9(MMP-9)(P<0.01), but also significantly reduced the expression of p-c-Src, PI3K, as well as p-Akt in vitro osteoclasts(P<0.01). In summary, bulleyaconitine A may inhibit RA bone destruction by regulating the Src/PI3K/Akt signaling pathway. This study provides experimental support for the treatment of RA bone destruction with bulleyaconitine A and lays a foundation for the clinical application of bulleyaconitine A.
Subject(s)
Aconitine/analogs & derivatives , Arthritis, Experimental , Arthritis, Rheumatoid , Drugs, Chinese Herbal , Animals , Proto-Oncogene Proteins c-akt/genetics , Phosphatidylinositol 3-Kinases/genetics , Molecular Docking Simulation , Signal Transduction , Arthritis, Rheumatoid/drug therapy , Arthritis, Rheumatoid/genetics , RNA, Messenger , Drugs, Chinese Herbal/pharmacologyABSTRACT
Aconitine is the main active component of Aconitum plants. Although aconitine has effects that include strengthening the heart, analgesia, anti-tumor, and immune-regulating effects, aconitine has both efficacy and toxicity, especially cardiotoxicity. Severe effects can include arrhythmia and cardiac arrest, which limits the clinical application of aconitine-containing traditional Chinese medicine. Ginsenoside Rb1(Rb1) is mainly found in plants, such as ginseng and Panax notoginseng, and has cardiovascular-protective and anti-arrhythmia effects. This study aimed to investigate the detoxifying effects of Rb1 on aconitine cardiotoxicity and the electrophysiological effect of Rb1 on aconitine-induced arrhythmia in rats. Pathological analysis, myocardial enzymatic indexes, and Western blotting were used to investigate the ameliorating effect of Rb1 on aconitine cardiotoxicity. Optical mapping was used to evaluate the effect of Rb1 on action potential and calcium signaling after aconitine-induced arrhythmia. Rb1 inhibited pathological damage caused by aconitine, decreased myocardial enzyme levels, and restored the balance of apoptotic protein expression by reducing the expression of Bax and cleaved caspase 3 and increasing the expression of Bcl-2, thereby reducing myocardial damage caused by aconitine. Rb1 also reduced the increase in heart rate caused by aconitine, accelerated action potential conduction and calcium signaling, and reduced the dispersion of action potential and calcium signal conduction. Rb1 reduced the cardiotoxicity of aconitine by attenuating aconitine-induced myocardial injury and inhibiting the aconitine-induced retardation of ventricular action potential and calcium signaling in rats.
Subject(s)
Aconitine , Calcium Signaling , Cardiotoxicity , Ginsenosides , Animals , Ginsenosides/pharmacology , Aconitine/analogs & derivatives , Cardiotoxicity/prevention & control , Rats , Calcium Signaling/drug effects , Male , Action Potentials/drug effects , Rats, Sprague-Dawley , Arrhythmias, Cardiac/chemically induced , Arrhythmias, Cardiac/prevention & control , Myocardium/metabolism , Myocardium/pathologyABSTRACT
Aconitum spp. are important medicinal plants mentioned in Ayurveda as Ativisa or Vatsanabha. The present study aims to evaluate anti-rheumatic potential in seven Aconitum species and correlation with aconitine and hypaconitine content. Anti-rheumatic potential was analyzed through inâ vitro xanthine oxidase inhibition, anti-inflammatory and ROS scavenging assays; and quantification of aconitine and hypaconitine with RP-HPLC method validated as per ICH guidelines. The findings reveal that A. palmatum possessed the most promising response (IC50 =12.68±0.15â µg/ml) followed by A. ferox (IC50 =12.912±1.87â µg/ml) for xanthin oxidase inhibition. We observed a wide variation in aconitine and hypaconitine content ranging from 0.018 %-1.37 % and 0.0051 %-0.077 % respectively on dry weight basis. Aconitine and hypaconitine showed moderate positive correlation (r=0.68 and 0.59 respectively) with anti-rheumatic potential. The study identifies potential alternative species of Aconitum that can help in sustainable availability of quality raw material.
Subject(s)
Aconitine/analogs & derivatives , Aconitum , Drugs, Chinese Herbal , Aconitine/pharmacology , Aconitine/analysis , Sikkim , Himalayas , Chromatography, High Pressure Liquid/methods , IndiaABSTRACT
This study aims to investigate the therapeutic effects of alkaloids in Tibetan medicine Bangna(Aconiti Penduli et Aconiti Flavi Radix) on osteoarthritis(OA) rats in vitro and in vivo and the underlying mechanisms. Chondrocytes were isolated from 2-3 week-old male SD rats and lipopolysaccharide(LPS) was used to induce OA in chondrocytes in vitro. Methyl thiazolyl tetrazolium(MTT) assay was used to investigate the toxicity of seven alkaloids(12-epi-napelline, songorine, benzoylaconine, aconitine, 3-acetylaconitine, mesaconitine, and benzoylmesaconine) to chondrocytes. Chondrocytes were classified into the control group, model group(induced by LPS 5 µg·mL~(-1) for 12 h), and administration groups(induced by LPS 5 µg·mL~(-1) for 12 h and incubated for 24 h). The protein expression of inflammatory factors cyclooxygenase-2(COX-2), inducible nitric oxide synthetase(iNOS), tumor necrosis factor-α(TNF-α), and interleukin-1ß(IL-1ß) in each group were detected by Western blot, and the protein expression of matrix metalloprotease-13(MMP-13), aggrecan, collagen â ¡, fibroblast growth factor 2(FGF2) by immunofluorescence staining. For the in vivo experiment, sodium iodoacetate was used to induce OA in rats, and the expression of MMP-13, TNF-α, and FGF2 in cartilage tissues of rats in each group was detected by immunohistochemistry. The results showed that the viability of chondrocytes could reach more than 90% under the treatment of the seven alkaloids in a certain dose range. Aconitine, 12-epi-napelline, songorine, 3-acetylaconitine, and mesaconitine could decrease the protein expression of inflammatory factors COX-2, iNOS, TNF-α and IL-1ß compared with the model group. Moreover, 12-epi-napelline, aconitine, and mesaconitine could down-regulate the expression of MMP-13 and up-regulate the expression of aggrecan and collagen â ¡. In addition, compared with the model group and other Bangna alkaloids, 12-epi-napelline significantly up-regulated the expression of FGF2. Therefore, 12-epi-napelline was selected for the animal experiment in vivo. Immunohistochemistry results showed that 12-epi-napelline could significantly reduce the expression of MMP-13 and TNF-α in cartilage tissues, and up-regulate the expression of FGF2 compared with the model group. In conclusion, among the seven Bangna alkaloids, 12-epi-napelline can promote the repair of OA in rats by down-regulating the expression of MMP-13 and TNF-α and up-regulating the expression of FGF2.
Subject(s)
Aconitine , Aconitum , Alkaloids , Medicine, Tibetan Traditional , Osteoarthritis , Aconitine/analogs & derivatives , Aconitine/therapeutic use , Aconitum/chemistry , Aggrecans/metabolism , Alkaloids/therapeutic use , Animals , Cells, Cultured , Cyclooxygenase 2/genetics , Cyclooxygenase 2/metabolism , Fibroblast Growth Factor 2/genetics , Fibroblast Growth Factor 2/therapeutic use , Interleukin-1beta/metabolism , Iodoacetic Acid/therapeutic use , Lipopolysaccharides , Male , Matrix Metalloproteinase 13/metabolism , NF-kappa B/metabolism , Osteoarthritis/drug therapy , Rats , Rats, Sprague-Dawley , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolismABSTRACT
This paper aims to study the difference in the intestinal absorption kinetics of main active components of Sini decoction and its separated recipes and explain the scientificity and rationality of the compatibility of Sini Decoction. A in situ intestinal perfusion rat model was established to evaluate the differences in the absorption of benzoylmesaconine, benzoylaconine, benzoylhypacoitine, mesaconitine, hypaconitine, glycyrrhizic acid, liquiritin and 6-gingerol from Sini Decoction and its separated recipes in the duodenum, jejunum and ileum by high performance liquid chromatography(HPLC). The results indicated that the Sini Decoction group was superior to the Aconiti Lateralis Radix Praeparata group in terms of absorption degree and rate for aconitum alkaloids. The absorption of benzoylmesaconine and hypaconitine in the duodenum, jejunum and ileum was faster and stronger in the Sini Decoction group(P<0.05). The absorption degree of glycyrrhizic acid in the duodenum was significantly higher in the Sini Decoction group than in the Glycyrrhizae Radix et Rhizoma group and the Glycyrrhizae Radix et Rhizoma-Zingiberis Rhizoma group(P<0.05). The absorption rate and degree of 6-gingerol in the ileum in the Sini Decoction group were significantly higher than those in the Zingiberis Rhizoma group(P<0.05). In short, Zingiberis Rhizoma and Glycyrrhizae Radix et Rhizoma can promote the absorption of aconitum alkaloids in different intestinal segments, which reflects the scientific composition of Sini Decoction.
Subject(s)
Aconitum , Alkaloids , Drugs, Chinese Herbal , Aconitine/analogs & derivatives , Animals , Catechols , Fatty Alcohols , Glycyrrhizic Acid , Intestinal Absorption , Kinetics , RatsABSTRACT
Mesaconitine (MA), one of the main diterpenoid alkaloids in Aconitum, has a variety of pharmacological effects, such as analgesia, anti-inflammation and relaxation of rat aorta. However, MA is a highly toxic ingredient. At present, studies on its toxicity are mainly focused on the heart and central nervous system, and there are few reports on the hepatotoxic mechanism of MA. Therefore, we evaluated the effects of MA administration on liver. SD rats were randomly divided into a normal saline (NS) group, a low-dose MA group (0.8 mg/kg/day) and a high-dose MA group (1.2 mg/kg/day). After 6 days of administration, the toxicity of MA on the liver was observed. Metabolomic and network toxicology methods were combined to explore the effect of MA on the liver of SD rats and the mechanism of hepatotoxicity in this study. Through metabonomics study, the differential metabolites of MA, such as L-phenylalanine, retinyl ester, L-proline and 5-hydroxyindole acetaldehyde, were obtained, which involved amino acid metabolism, vitamin metabolism, glucose metabolism and lipid metabolism. Based on network toxicological analysis, MA can affect HIF-1 signal pathway, MAPK signal pathway, PI3K-Akt signal pathway and FoxO signal pathway by regulating ALB, AKT1, CASP3, IL2 and other targets. Western blot results showed that protein expression of HMOX1, IL2 and caspase-3 in liver significantly increased after MA administration (p < 0.05). Combined with the results of metabonomics and network toxicology, it is suggested that MA may induce hepatotoxicity by activating oxidative stress, initiating inflammatory reaction and inducing apoptosis.
Subject(s)
Chemical and Drug Induced Liver Injury , Phosphatidylinositol 3-Kinases , Aconitine/analogs & derivatives , Animals , Chemical and Drug Induced Liver Injury/etiology , Interleukin-2 , Rats , Rats, Sprague-Dawley , Rats, WistarABSTRACT
Objective: To investigate the analgesic efficacy and safety of state-dependent sodium channel blocker-bulleyaconitine combined with calcium channel blocker-gabapentin on postherpetic neuralgia (PHN). Methods: A double-blind, randomized, placebo-controlled, parallel-group, multi-center study involving Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Qinghai Provincial People's Hospital, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine Affiliated to Shanghai University of Traditional Chinese Medicine, the Second Affiliated Hospital of Kunming Medical University was performed from September 2018 to December 2019. A total of 75 PHN patients were randomly divided into the experiment group (n=41) and the control group (n=34). On the basis of first-line treatment with gabapentin, the experiment group was given bulleyaconitine A tablets, while the control group was given placebo. The primary outcome was a 50% improvement in the visual analogue scale (VAS), and the effective rate of achieving the primary outcome between the two groups was compared; the Cox regression model was used to analyze the impact of related factors on the disease outcome. Secondary outcomes including scores of pain scales (ID-pain, DN4), Patient Health Questionnaire (PHQ-9), 7-item Generalized Anxiety Disorder (GAD-7) at 1, 2, 3, 4, 8, 12 weeks after treatment were applied to evaluate the efficacy and safety of the combination of bulleyaconitine A tablets with first-line drug in the treatment of PHN. Results: The effective rate was 68.3% (28/41) and the time reached the primary outcome was 28 (7, 84) days in the experiment group, while in the control group, the effective rate was 52.9% (18/34) and the time reached the primary outcome was 56 (14, 84) days. Cox regression analysis indicated that the grouping factor of oral bulleyaconitine A tablets was an independent factor for improving the outcome of PHN (HR=2.063, 95%CI: 1.059-4.018, P<0.05), and the probability of the experiment group reaching the primary outcome was 2.063 times that of the control group (P<0.05). Meanwhile, the outcome probability of the long disease course group (>6 months) was only 0.201 times that of the short disease course group (<6 months) (HR=0.201, 95%CI: 0.073-0.551, P<0.05). There was no statistically significant difference in the trend of VAS between the two groups (P>0.05). The scores of ID-pain, DN4, PHQ-9, and GAD-7 of the two groups were significantly improved compared with those before enrollment (P<0.05), but the differences between the two groups were not statistically significant (all P>0.05). Conclusion: Bulleyaconitine A tablet can promote the therapeutic efficacy of gabapentin, and improve the outcome of PHN in a short period of 3 months.
Subject(s)
Neuralgia, Postherpetic , Aconitine/analogs & derivatives , Analgesics/therapeutic use , China , Double-Blind Method , Gabapentin , Humans , Treatment OutcomeABSTRACT
OBJECTIVES: Identifying drugs with time-varying efficacy or toxicity, and understanding the underlying mechanisms would help to improve treatment efficacy and reduce adverse effects. In this study, we uncovered that the therapeutic effect of Fuzi (the lateral root of Aconitum carmichaelii Debeaux) depended on the dosing time in mice with adenine-induced chronic kidney disease (CKD). METHODS: The Fuzi efficacy was determined by biomarker measurements [i.e. plasma creatinine (CRE), blood urea nitrogen (BUN) and urinary N-acetyl-ß-D-glucosaminidase (NAG)], as well as inflammation, fibrosis and histological analyses. Circadian regulation of Fuzi pharmacokinetics and efficacy was evaluated using brain and muscle Arnt-like protein-1 (Bmal1)-deficient (Bmal1-/-) mice. KEY FINDINGS: The Fuzi efficacy was higher when the drug was dosed at ZT10 and was lower when the drug was dosed at other times (ZT2, ZT6, ZT14, ZT18 and ZT22) according to measurements of plasma CRE, BUN and urinary NAG. Consistently, ZT10 (5 PM) dosing showed a stronger protective effect on the kidney (i.e. less extensive tubular injury) as compared to ZT22 (5 AM) dosing. This was supported by lower levels of inflammatory and fibrotic factors (IL-1ß, IL-6, Tnf-α, Ccl2, Tgfb1 and Col1a1) at ZT10 than at ZT22. Pharmacokinetic analyses showed that the area under the curve (AUC) values (reflective of systemic exposure) and renal distribution of aconitine, hypaconitine and mesaconitine (three putative active constituents) for Fuzi dosing at ZT10 were significantly higher than those for herb dosing at ZT22, suggesting a role of circadian pharmacokinetics in Fuzi chronoefficacy. Drug efficacy studies confirmed that aconitine, hypaconitine and mesaconitine possessed a kidney-protecting effect. In addition, genetic knockout of Bmal1 in mice abolished the time-dependency of Fuzi pharmacokinetics and efficacy. This reinforced the existence of chronoefficacy for Fuzi and supported the role of circadian pharmacokinetics in Fuzi chronoefficacy. CONCLUSIONS: The efficacy of Fuzi against CKD depends on the dosing time in mice, which is associated with circadian pharmacokinetics of the three main active constituents (i.e. aconitine, hypaconitine and mesaconitine). These findings highlight the relevance of dosing time in the therapeutic outcomes of herbal medicines.
Subject(s)
Chronopharmacokinetics , Diterpenes , Drugs, Chinese Herbal , Renal Insufficiency, Chronic , ARNTL Transcription Factors/genetics , Aconitine/analogs & derivatives , Aconitine/analysis , Alkaloids/administration & dosage , Alkaloids/pharmacokinetics , Animals , Anti-Inflammatory Agents/administration & dosage , Anti-Inflammatory Agents/pharmacokinetics , Diterpenes/administration & dosage , Diterpenes/pharmacokinetics , Dose-Response Relationship, Drug , Drugs, Chinese Herbal/administration & dosage , Drugs, Chinese Herbal/pharmacokinetics , Kidney Function Tests/methods , Mice , Mice, Knockout , Plant Roots , Protective Agents/administration & dosage , Protective Agents/pharmacokinetics , Renal Insufficiency, Chronic/drug therapy , Renal Insufficiency, Chronic/metabolism , Treatment OutcomeABSTRACT
In this paper, UPLC-MS/MS was used to determine 8-deacetyl-yunaconitine in the plasma of rats after oral and intravenous administration. Six rats were orally (po) administered 8-deacetyl-yunaconitine (5 mg/kg), while another six rats were intravenously (iv) administered the drug (0.1 mg/kg). A standard curve of known concentrations of 8-deacetyl-yunaconitine in rat plasma was generated over the range of 0.3-600 ng/mL. The intra-day and inter-day precision of 8-deacetyl-yunaconitine in rat plasma was lower than 15 %, while the accuracy ranged between 97.7 % and 105.5 %. In addition, the matrix effect ranged between 95.3 % and 105.6 %, while the recovery was greater than 82.8 %. This determined method was then applied in measuring the pharmacokinetics of 8-deacetyl-yunaconitine in rats. The AUC(0-t) values were 73.0 ± 24.6 and 1770.0 ± 530.6 ng/mL h for intravenous and oral administration, respectively, and the bioavailability was 48.5 %. The half-life (t1/2) was determined to be 4.5 ± 1.7 h and 3.2 ± 0.7 h for intravenous and oral administration, respectively, indicating that the metabolism of the xenobiotic was quick in the rats.
Subject(s)
Drugs, Chinese Herbal , Tandem Mass Spectrometry , Aconitine/analogs & derivatives , Administration, Oral , Animals , Chromatography, High Pressure Liquid , Chromatography, Liquid , Rats , Rats, Sprague-Dawley , Reproducibility of ResultsABSTRACT
The lateral roots of Aconitum carmichaelii, known in Chinese as fuzi, are officially recognized as a materia medica in the Chinese Pharmacopoeia and used culinarily to prepare herbal soups. A strategy combining UPLC-qToF-MS analysis of A. carmichaelii and its intraspecies and interspecies chemometrics study was developed to examine the distribution of Aconitum marker metabolites. Four diterpenoid alkaloids were recognized to be important markers in fuzi, and another 15 markers were identified to differentiate A. carmichaelii from adulterant species. The detected fuzi markers, mesaconitine (47) and hypaconitine (51), are known to be the principal toxins in this herb, while fuziline (6) and benzoylmesaconine (25) are associated with its medicinal properties. Additional marker compounds have been detected in other Aconitum species that are useful for identifying adulteration. This study provides a useful resource for detecting traditional Chinese medicine (TCM) adulterants and assisting in the quality control of botanical products in TCM and beyond.
Subject(s)
Aconitum/chemistry , Alkaloids/analysis , Diterpenes/analysis , Drugs, Chinese Herbal/analysis , Aconitine/analogs & derivatives , Aconitine/analysis , Aconitum/classification , China , Chromatography, Liquid , Drug Contamination , Mass Spectrometry , Medicine, Chinese Traditional , Molecular Structure , Plant Roots/chemistryABSTRACT
Mesaconitine, one of Aconitum carmichaelii Debx bioactive compounds, was shown to evoke Ca2+ homeostasis and its related physiological effects in endothelial cell types. However, the effect of mesaconitine on Ca2+ signaling and cell viability in human brain microvascular endothelial cells is unclear. This study focused on exploring whether mesaconitine changed cytosolic Ca2+ concentrations ([Ca2+]i), affected cell viability, and established the relationship between Ca2+ signaling and viability in HBEC-5i human brain microvascular endothelial cells. In HBEC-5i cells, cell viability was measured by the cell proliferation reagent (WST-1). [Ca2+]i was measured by the Ca2+-sensitive fluorescent dye fura-2. Mesaconitine (10-100 µM) concentration dependently induced [Ca2+]i rises. Ca2+ removal reduced the signal by approximately 25%. Mesaconitine (40-100 µM) caused cytotoxicity in HBEC-5i cells. This cytotoxic response was significantly reversed by chelation of cytosolic Ca2+ with BAPTA/AM. In Ca2+-containing medium, mesaconitine-induced Ca2+ entry was inhibited by 25% by modulators of store-operated Ca2+ channels and protein kinase C (PKC). Furthermore, mesaconitine also induced Mn2+ influx suggesting of Ca2+ entry. In Ca2+-free medium, treatment with the endoplasmic reticulum Ca2+ pump inhibitor thapsigargin abolished mesaconitine-evoked [Ca2+]i rises. Conversely, treatment with mesaconitine abolished thapsigargin-evoked [Ca2+]i rises. Inhibition of phospholipase C (PLC) with U73122 abolished mesaconitine-induced [Ca2+]i rises. In sum, mesaconitine caused cytotoxicity that was triggered by preceding [Ca2+]i rises. Furthermore, mesaconitine induced [Ca2+]i rises by evoking Ca2+ entry via PKC-sensitive store-operated Ca2+ channels and PLC-dependent Ca2+ release from the endoplasmic reticulum. It suggests that Ca2+ signaling have a potential cytotoxic effect on mesaconitine-treated human brain microvascular endothelial cells.
Subject(s)
Aconitine/analogs & derivatives , Calcium Signaling/drug effects , Endothelial Cells/drug effects , Endothelial Cells/metabolism , Aconitine/administration & dosage , Aconitum , Cell Line , Cell Survival/drug effects , Humans , Plant Extracts/toxicityABSTRACT
BACKGROUND/AIMS: Acute pancreatitis-associated lung injury (APALI) is one of the most common and most dangerous form of extra-pancreatic organ damage in severe acute pancreatitis (SAP). The treatment options for SAP were limited thus far; as a result, approximately 60%-80% of patients with SAP would die within a week. Hypaconitine (HC), one of the most important active ingredients in a Mongolian traditional medicine Radix Aconiti Kusnezoffii has an excellent anti-inflammatory effect. MATERIALS AND METHODS: To ascertain whether HC has a protective effect against APALI, we investigated the therapeutic effects and the underlying mechanisms in vivo and in vitro and attempted to elucidate the mechanism in detail. In this study, APALI rats and human pulmonary microvascular endothelial cells were treated with therapeutic doses of HC after establishing a model with sodium taurocholate and lipopolysaccharide, respectively. RESULTS: Serum amylase and lipase activity, lung wet/dry weight ratio, lung myeloperoxidase activity, and pancreatic and lung histopathological changes showed that HC alleviated APALI in a dose-dependent way, which can be abolished by an aquaporin-1 (AQP-1) knockdown. The results of the reverse transcriptase polymerase chain reaction, Western blot, and immunohistochemical staining confirmed the expression of AQP-1, a kind of transmembrane protein that mainly distributed in the membranes of pulmonary cells and contributed to maintain water balance in the body by interacting with tumor necrosis factor-alpha (TNF-α), is negatively associated with APALI. On the contrary, HC treatment up-regulated AQP-1 expression and down-regulated the TNF-α expression as a consequence in APALI. CONCLUSION: These results suggest that HC has a good anti-inflammatory therapeutic effect on APALI with a possible underlying mechanism that affects the AQP-1/TNF-α pathway.
Subject(s)
Aconitine/analogs & derivatives , Acute Lung Injury/drug therapy , Anti-Inflammatory Agents/pharmacology , Aquaporin 1/metabolism , Pancreatitis/drug therapy , Tumor Necrosis Factor-alpha/metabolism , Aconitine/pharmacology , Acute Disease , Acute Lung Injury/etiology , Animals , Dose-Response Relationship, Drug , Endothelial Cells/metabolism , Humans , Lung/drug effects , Lung/pathology , Male , Microvessels/cytology , Pancreas/pathology , Pancreatitis/complications , Rats , Rats, Sprague-Dawley , Respiratory Mucosa/blood supply , Signal Transduction/drug effects , Up-Regulation/drug effectsABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Goshajinkigan (GJG), a traditional Japanese Kampo formula, has been shown to exhibit several pharmacological actions, including antinociceptive effects. Processed aconite root (PA), which is considered to be an active ingredient of GJG, has also been demonstrated to have an ameliorative effect on pain, such as diabetic peripheral neuropathic pain. We recently identified neoline as the active ingredient of both GJG and PA that is responsible for its effects against oxaliplatin-induced neuropathic pain in mice. AIM OF THE STUDY: In the present study, we investigated whether GJG, PA, and neoline could inhibit Nav1.7 voltage-gated sodium channel (VGSC) current and whether neoline could ameliorate mechanical hyperalgesia in diabetic mice. MATERIALS AND METHODS: To assess the electrophysiological properties of GJG extract formulation, powdered PA, and neoline on Nav1.7 VGSCs, whole-cell patch clamp recording was performed using human HEK293 cells expressing Nav1.7 VGSCs. In addition, the ameliorative effects of neoline on diabetic peripheral neuropathic pain were evaluated using the von Frey test in streptozotocin (STZ)-induced diabetic model mice. RESULTS: GJG extract formulation significantly inhibited Nav1.7 VGSC peak current. Powdered PA also inhibited Nav1.7 VGSC peak current. Like GJG and PA, neoline could inhibit Nav1.7 VGSC current. When diabetic mice were treated with neoline by intraperitoneal acute administration, the mechanical threshold was increased in diabetic mice, but not in non-diabetic mice, in a behavioral study. CONCLUSION: These results suggest that neoline might be a novel active ingredient of GJG and PA that is one of responsible ingredients for ameliorating mechanical hyperalgesia in diabetes via the inhibition of Nav1.7 VGSC current at least.
Subject(s)
Aconitine/analogs & derivatives , Aconitum , Analgesics/pharmacology , Diabetic Neuropathies/prevention & control , Drugs, Chinese Herbal/pharmacology , Hyperalgesia/prevention & control , NAV1.7 Voltage-Gated Sodium Channel/drug effects , Plant Roots , Voltage-Gated Sodium Channel Blockers/pharmacology , Aconitine/isolation & purification , Aconitine/pharmacology , Aconitum/chemistry , Analgesics/isolation & purification , Animals , Behavior, Animal/drug effects , Diabetes Mellitus, Experimental/complications , Diabetic Neuropathies/etiology , Diabetic Neuropathies/metabolism , Diabetic Neuropathies/physiopathology , Drugs, Chinese Herbal/isolation & purification , HEK293 Cells , Humans , Hyperalgesia/etiology , Hyperalgesia/metabolism , Hyperalgesia/physiopathology , Male , Membrane Potentials , Mice, Inbred ICR , NAV1.7 Voltage-Gated Sodium Channel/genetics , NAV1.7 Voltage-Gated Sodium Channel/metabolism , Pain Threshold/drug effects , Plant Roots/chemistry , Voltage-Gated Sodium Channel Blockers/isolation & purificationABSTRACT
Context: It is common sense that chewing a mint leaf can cause a cooling feeling, while chewing ginger root will produce a burning feeling. In Traditional Chinese Medicine (TCM), this phenomenon is referred to as 'cold/hot' properties of herbs. Herein, it is reported that TCM with different "cold/hot" properties have different effects on the variation of cells.Objective: To explore the intrinsic 'cold/hot' properties of TCM from the perspective of cellular and molecular biology.Materials and methods: A375 cells were selected using Cancer Cell Line Encyclopaedia (CCLE) analysis and western blots. Hypaconitine and baicalin were selected by structural similarity analysis from 56 and 140 compounds, respectively. A wireless thermometry system was used to measure cellular temperature change induced by different compounds. Alteration of intracellular calcium influx was investigated by means of calcium imaging.Results: The IC50 values of GSK1016790A, HC067047, hypaconitine, and baicalin for A375 cells are 8.363 nM, 816.4 µM, 286.4 µM and 29.84 µM, respectively. And, 8 µM hypaconitine induced obvious calcium influx while 8 µM baicalin inhibited calcium influx induced by TRPV4 activation. Cellular temperature elevated significantly when treated with GSK1016790A or hypaconitine, while the results were reversed when cells were treated with HC067047 or baicalin.Discussion and conclusions: The changes in cellular temperature are speculated to be caused by the alteration of intracellular calcium influx mediated by TRPV4. In addition, the 'cold/hot' properties of compounds in TCM can be classified by using cellular temperature detection.
Subject(s)
Cell Proliferation/drug effects , Drugs, Chinese Herbal/pharmacology , Keratinocytes/drug effects , Thermogenesis/drug effects , Aconitine/analogs & derivatives , Aconitine/pharmacology , Calcium/metabolism , Cell Line, Tumor , Cell Survival/drug effects , Cold Temperature , Flavonoids/pharmacology , Gene Expression/drug effects , Hot Temperature , Humans , Leucine/analogs & derivatives , Leucine/pharmacology , Medicine, Chinese Traditional/methods , Morpholines/pharmacology , Pyrroles/pharmacology , Sulfonamides/pharmacology , TRPV Cation Channels/genetics , TRPV Cation Channels/metabolismABSTRACT
Ginseng and aconite are well-known couplet medicinals. Ginsenoside Rg1 is the main active ingredient in ginseng, and aconitine (AC), benzoylaconine (BAC) and aconine (ACN) are three representative alkaloids in aconite, which belong to the diester alkaloids, monoester alkaloids and alkanolamine alkaloids respectively. The aim of this study was to investigate the pharmacokinetic effects of ginsenoside Rg1 on the three types of alkaloids and to provide evidences for their compatibility mechanism. In this study, the ginsenoside Rg1 was simultaneously intragastrically administered to rats with AC, BAC and ACN, respectively, and the rat plasma was collected at different time points. The plasma drug concentrations of the three types of alkaloids were determined by UHPLC-MS/MS, and the pharmacokinetic parameters were calculated. The results indicated that the peak concentration and area under the concentration-time curve of BAC were significantly increased (P < 0.05), those for AC were decreased (P < 0.05), and the values for ACN did not change after pretreatment with ginsenoside Rg1. It was inferred that ginsenoside Rg1 may affect the absorption and metabolism of AC and BAC and then change their pharmacokinetic parameters. Subsequently, their absorption and metabolism were further investigated using the Caco-2 cell monolayer and rat liver microsomes in vitro. The Caco-2 cell monolayer absorption assay indicated that ginsenoside Rg1 could promote the absorption of AC and BAC, and the rat liver microsomes metabolism assay indicated that ginsenoside Rg1 accelerated the metabolism of AC and did not affect the other two alkaloids. All of the results indicated that ginsenoside Rg1 may reduce the toxicity of aconite and improve its efficacy by promoting the absorption of BAC and accelerating the metabolism of AC. These results could provide evidence for the compatibility mechanism of the traditional Chinese herbal formula Shenfu Decoction.
Subject(s)
Aconitine/analogs & derivatives , Aconitine/pharmacokinetics , Chromatography, High Pressure Liquid/methods , Ginsenosides/pharmacokinetics , Tandem Mass Spectrometry/methods , Aconitine/administration & dosage , Aconitine/blood , Administration, Oral , Animals , Caco-2 Cells , Ginsenosides/administration & dosage , Ginsenosides/blood , Humans , Linear Models , Male , Microsomes, Liver/metabolism , Rats , Rats, Sprague-Dawley , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
Lappaconitine (LA), a potent analgesic drug extracted from the root of natural aconitum species, has been clinically used for years because of its effectiveness and non-addictive properties. However, it is mainly limited in oral and intravenous administration in the form of Lappaconitine Hydrobromide (LAH). In this work, Lappaconitine trifluoroacetate (LAF), a new derivative of LA, was successfully obtained by introducing organofluorine group to LA. This new compound had a lower toxicity (LD50 of 21.14 mg·kg-1), improved analgesic effect and longer half-life (T1/2 of 2.24 h) when compared with LAH. Moreover, in vitro transdermal permeation (Jss of 206.82 µg·cm-2·h-1) of LAF was 30.54% higher than that of LAH, means that LAF can be conveniently used for transdermal drug delivery (TDD). Therefore, drug membranes with PVA solution (10 wt%) containing LAF in various amounts were fabricated by electrospinning. The in vitro release tests confirmed that up to 81.43% of LAF in the PVA/LAF nanofibrous membranes could be released in 72 h, accompanied by significant analgesic effect when compared with the blank control group. In conclusion, the prepared LAF-loaded membrane is a novel formulation for the treatment of chronic and long-term pain.
Subject(s)
Aconitine/analogs & derivatives , Analgesics/administration & dosage , Nanofibers/chemistry , Polyvinyl Alcohol/administration & dosage , Trifluoroacetic Acid/administration & dosage , Aconitine/administration & dosage , Aconitum/chemistry , Administration, Cutaneous , Analgesics/pharmacology , Animals , Drug Delivery Systems , In Vitro Techniques , Magnetic Resonance Spectroscopy , Male , Materials Testing , Mice , Mice, Inbred BALB C , Microscopy, Electron, Scanning , Pain Management/methods , Photoelectron Spectroscopy , Plant Extracts/pharmacology , Rats , Rats, Sprague-Dawley , Skin/drug effects , Solubility , Spectroscopy, Fourier Transform InfraredABSTRACT
BACKGROUND: Therapeutic applications of Fuzi (lateral root of Aconitum carmichaeli Debx) are seriously concerned with its toxic effects. Strategies and approaches to reducing toxicity are of great interest. PURPOSE: We aimed to characterize the diurnal rhythm of Fuzi toxicity, and to determine the role of metabolism and pharmacokinetics in generating toxicity rhythmicity. METHODS: Toxicity was determined based on assessment of heart injury and animal survival after dosing mice with Fuzi decoction at different circadian time points. Circadian clock control of pharmacokinetics and toxicity was investigated using Bmal1-deficient (Bmal1-/-) mice. RESULTS: Fuzi exhibited a diurnal rhythmicity in cardiotoxicity (reflected by plasma CK-MB and LDH levels). The highest level of toxicity was observed at ZT10 (5 PM), while the lowest level of toxicity occurred at ZT22 (5 AM). Also, a higher mortality rate was observed at ZT10 and lower mortality rates at other times of the day. ZT10 dosing of Fuzi generated higher systemic exposures of three toxic alkaloid ingredients aconitine (AC), hypaconitine (HA) and mesaconitine (MA) compared to ZT22. This was accompanied by reduced the formation of the metabolites (N-deethyl-AC, didemethyl-HA and 2hydroxylMA) at ZT10. Bmal1 ablation resulted in an increased level of Fuzi toxicity at ZT22, while having no influences when drug was dosed at ZT10. As a consequence, circadian time-dependent toxicity of Fuzi was lost in Bmal1-deficient mice. In addition, Bmal1 ablation increased the plasma concentrations of AC, HA and MA in mice after oral gavage of Fuzi, and reduced formation of their metabolites (N-deethyl-AC, didemethyl-HA and 2hydroxylMA). Moreover, Fuzi metabolism in wild-type liver microsomes was more extensive at ZT22 than at ZT10. Bmal1 ablation abrogated circadian time-dependency of hepatic Fuzi metabolism. CONCLUSIONS: Fuzi chronotoxicity in mice was attributed to time-varying hepatic metabolism and systemic exposure regulated by circadian clock. The findings may have implications in reducing Fuzi toxicity with a chronotherapeutic approach.
Subject(s)
Aconitum/chemistry , Circadian Clocks/drug effects , Plant Extracts/pharmacokinetics , Plant Extracts/toxicity , ARNTL Transcription Factors/genetics , Aconitine/analogs & derivatives , Aconitine/pharmacokinetics , Animals , Chromatography, High Pressure Liquid/methods , Circadian Clocks/physiology , Diterpenes , Drugs, Chinese Herbal , Mice , Mice, Inbred C57BL , Mice, Mutant Strains , Microsomes, Liver/drug effects , Toxicity Tests/methodsABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: The herbs of Aconitum are the essential Traditional Chinese medicine and have played an indispensable role in many Asian countries for thousands of years to treat critical illnesses, and chronic, stubborn diseases. However, Aconitum may induce severe neurotoxicity and even death. So far the mechanism of Aconitum penetrating the blood-brain barrier (BBB) is still unclear. AIM OF THE STUDY: To determine whether influx transporters contribute to the brain uptake of the highly toxic alkaloids in Aconitum including aconitine (AC), mesaconitine (MA) and hypaconitine (HA). MATERIALS AND METHODS: The uptake of AC, MA and HA was characterized using in vitro hCMEC/D3 model and in situ mouse brain perfusion. In hCMEC/D3 cells, the effect of incubation temperature, time, initial drug concentration, energy (NaN3), extracellular and intracellular pH (FCCP and NH4Cl), the prototypical substrates/inhibitors of known organic cation transporting carriers and trans-stimulation (pre-incubating with pyrilamine and diphenhydramine) on the cellular uptake were studied. In addition, the effect of silencing OCTN1, OCTN2 and PMAT by specific siRNA was investigated. In mice, the contribution of the proton-coupled antiporter on the brain uptake of Aconitum was investigated by chemical inhibition. RESULTS: In hCMEC/D3 cells, AC, MA and HA were each taken up in a temperature-, time- and concentration-dependent manner, which were reduced by NaN3 and FCCP. Regulation of extracellular and intracellular pH as well as trans-stimulation studies showed that AC, MA and HA were transported by a proton-coupled antiporter expressed at the plasma membrane that could also transport pyrilamine and diphenhydramine. Each uptake was markedly inhibited by various cationic drugs, but insensitive to the prototypical substrates/inhibitors of identified organic cation transporting carriers, such as OCTs, PMAT, MATEs and OCTNs. In addition, silence of OCTN1, OCTN2 and PMAT had no significant inhibitory effect on the uptake of AC, MA and HA. In mice, the brain uptake of each alkaloid measured by in situ brain perfusion was suppressed by diphenhydramine when the transport capacity of P-gp/Bcrp at the BBB was chemically inhibited. CONCLUSIONS: A novel proton-coupled organic cation antiporter plays a predominant role in the blood to brain influx of AC, MA and HA at the BBB, and thus affect the safety of Aconitum species.
Subject(s)
Aconitine/analogs & derivatives , Aconitum , Antiporters/metabolism , Blood-Brain Barrier/metabolism , Organic Cation Transport Proteins/metabolism , Aconitine/pharmacology , Animals , Cell Line , Humans , Male , Mice, Inbred ICR , Organic Cation Transport Proteins/genetics , Protons , RNA, Small Interfering/geneticsABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Processed aconite root (PA), the root of Aconitum carmichaeli (Ranunculaceae), is a crude drug used in traditional Chinese or Japanese kampo medicine to treat pain associated with coldness. In our previous study, PA and its active ingredient, neoline, alleviated oxaliplatin-induced peripheral neuropathy in mice. AIM OF THE STUDY: The present study investigated the effects of PA on a murine peripheral neuropathy model induced by intraperitoneal injection of paclitaxel and partial ligation of the sciatic nerve (Seltzer model), and identified its active ingredients. MATERIALS AND METHODS: PA powder (1â¯g/kg/day) was orally administered, and either neoline or benzoylmesaconine (10â¯mg/kg/day) was subcutaneously injected into the murine model. Mechanical hyperalgesia was evaluated via the von Frey filament method. PA extract was orally administered to rats; blood samples were chronologically collected, and the plasma concentrations of Aconitum alkaloids were measured. The contents of Aconitum alkaloids in commercial PA products were also measured. RESULTS: PA extract and neoline significantly attenuated the mechanical hyperalgesia induced by either paclitaxel or partial ligation of the sciatic nerve in mice. In the plasma samples of rats treated with PA extract, higher concentrations of benzoylmesaconine and neoline were apparent among Aconitum alkaloids. The contents of benzoylmesaconine and neoline varied among PA products with different processing procedures. Subcutaneous injection of benzoylmesaconine did not attenuate the hyperalgesia induced by each paclitaxel, partial ligation of the sciatic nerve, or oxaliplatin in mice. CONCLUSIONS: The present results indicate that PA and its active ingredient, neoline, are promising agents for the alleviation of neuropathic pain. Neoline can be used as a marker compound to determine the quality of the PA products for the treatment of neuropathic pain.
Subject(s)
Aconitine/analogs & derivatives , Aconitum , Analgesics/therapeutic use , Hyperalgesia/drug therapy , Neuralgia/drug therapy , Peripheral Nerve Injuries/drug therapy , Aconitine/pharmacokinetics , Aconitine/therapeutic use , Analgesics/pharmacokinetics , Animals , Antineoplastic Agents, Phytogenic , Male , Mice , Neuralgia/chemically induced , Paclitaxel , Plant Roots , Rats, Wistar , Sciatic Nerve/injuriesABSTRACT
The unintentional ingestion of toxic compounds in herbs is not uncommon in many parts of the world. To provide timely and life-saving care in the emergency department, it is essential to develop a point-of-care analytical method that can rapidly identify these toxins in herbs. Since electrospray laser desorption ionization mass spectrometry (ELDI/MS) has been successfully used to characterize non-volatile chemical compounds without sample preparation, it was used to identify toxic herbal compounds in this study. The herbal toxins were collected either by sweeping a metallic probe across the surface of a freshly cut herb section or by directly sampling extracts of ground herbal powder. The analytes on the probe were then desorbed, ionized and detected using ELDI/MS, wherein analysis of the herbal toxins was completed within 30 s. This approach allows for the rapid morphological recognition of herbs and early point-of-care identification of herbal toxins for emergency management and is promising in providing important toxicological information to ensure appropriate medical treatment.