Effect of acute pancreatitis on the pharmacokinetics of Chinese herbal ointment Liu-He-Dan in anaesthetized rats

Ethnopharmacological relevance

Chinese herbal preparation of Liu-He-Dan ointment has been adapted for acute pancreatitis in external application for many years in West China.

Aim of the study

To investigate the effect of acute pancreatitis on the pharmacokinetics of Liu-He-Dan ointment in rats while it was used externally on belly.

Materials and methods

Twelve male Sprague-Dawley rats were randomly divided into acute pancreatitis model group (n=6) and normal group as a control (n=6). Chinese herbal Liu-He-Dan ointment was used externally on belly. Emodin, rhein, aloe emodin, physcion and chrysophanol in plasma and pancreas (at 48 h) were detected and quantified by liquid chromatography–tandem mass spectrometry. Amylase in plasma were determined with iodide process.

Results

Among the five components, only emodin, aloe emodin and physcion from Liu-He-Dan were detected in plasma and pancreas. The absorption of each component was tended to decrease in acute pancreatitis group after topically management with Liu-He-Dan ointment on rats’ abdomen. The T_max, C_max and area under curve (AUC) of each component were distinctly lower in AP group than those in normal group (p<0.05). However, the T_1/2α and mean retention time (MRT) of emodin lasted longer in acute pancreatitis group than those in normal group (p<0.05). There was no statistical difference in the MRT of aloe emodin and physcion between the two groups. Emodin could be detected in all rats’ pancreas at 48 h in both groups, while its mean pancreatic concentration was higher in acute pancreatitis model group than in normal group (0.91±0.68, 0.41±0.36, respectively). Physcion could be detected in pancreas of most acute pancreatitis models, but not in normal rats. Aloe emodin was found in all pancreas from acute pancreatitis models while only one in normal group. The level of amylase in Liu-He-Dan group was obviously lower than that in the AP model group (p=0.0055).

Conclusion

We concluded that acute pancreatitis may significantly affect the pharmacokinetics of Liu-He-Dan while external applied on belly, which indicated the dosage modification in AP. However, acute pancreatitis seems to promote the distribution of the detected components into pancreas. The ointment could help relieve the disease of pancreatitis.     

Metabolic profiling reveals therapeutic biomarkers of processed Aconitum Carmichaeli Debx in treating hydrocortisone induced Kidney-Yang deficiency syndrome rats

Ethnopharmacological relevance

Kidney-Yang Deficiency syndrome (KYDS) is a diagnostic pattern in traditional Chinese medicine (TCM) and clinical data showed that the unbalance in adrenal cortical hormone is the key issue in KYDS patients. The processed Ranunculaceae Aconitum carmichaeli debx (Bai-Fu-Pian in Chinese, BFP) is one of the most commonly used Chinese herbs for treating KYDS. The present study was conducted to explore the therapeutic biomarkers of the BFP in treating hydrocortisone administration induced KYDS rats.

Materials and methods

Thirty male Sprague-Dawley rats were randomly divided into five groups with six in each group. KYDS in rats was induced by i.p. injection of hydrocortisone at the dose of 10 mg/kg per day for 15 days as described previously. The rats with KYDS were administered orally, starting from the day of hydrocortisone administration stopped, with BFP extract at the dose of 0.32 g/kg, 0.64 g/kg and 1.28 g/kg per day respectively for 15 days. The blood samples were collected for the liquid chromatography quadruple time-of-flight mass spectrometry (LC-Q-TOF-MS) test, as well as radioimmunoassay to determine the concentrations of cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP) and adrenocorticotrophic hormone (ACTH). The metabolic responses to BFP administration were investigated by using the principal components analysis (PCA) and orthogonal partial least squares analysis (OPLS). Bioinformatics analyses were performed by using the Ingenuity Pathway Analysis (IPA). Variance analysis and linear regression analysis were used in this study.

Results

The signs and concentrations of cAMP, cGMP and ACTH in the model rats were similar to those previously described about KYDS rats and BFP treatment can reverse the changes. Seventeen significantly changed metabolites among different groups were identified. Thirteen metabolites were identified in the KYDS rats comparing to healthy rats with nine up-regulated and four down-regulated. After BFP treatment at three dosages, five up-regulated metabolites including phosphate, betaine, (4-hydroxyphenyl) acetaldehyde, 5-hydroxyindol-3-acetic acid and 5'-phosphoribosyl-N-formylglycinamide were dose-dependently reversed. The network analysis with IPA showed that four canonical pathways including superpathway of methionine degradation, purine nucleotides De Novo biosynthesis II, tyrosine synthesis and serotonin receptor signaling involved the therapeutic mechanism of BFP in treating the KYDS rats.

Conclusions

Five therapeutic biomarkers (phosphate, betaine, (4-hydroxyphenyl) acetaldehyde, 5-hydroxyindol-3-acetic acid and 5'-phosphoribosyl-N-formylglycinamide) and two corresponding canonical pathways (amino acid metabolism and purine nucleotide metabolism) were identified to be involved in the therapeutic mechanism of BFP treating the KYDS.     

Rapid-resolution liquid chromatography TOF-MS for urine metabolomic analysis of collagen-induced arthritis in rats and its applications

Aim of the study

Rheumatoid arthritis (RA) is a kind of autoimmune diseases characterized by persistent synovitis, systemic inflammation and autoantibodies. Huang-Lian-Jie-Du-Tang (HLJDT) is a traditional Chinese medicine (TCM) with anti-inflammatory activity. In the present study, a rapid-resolution liquid chromatography tandem time-of-flight mass spectrometry (RRLC-TOF-MS) based metabolomic study was developed to obtain a systematic view of the progression of RA and assess the efficacy of HLJDT and its components in collagen-induced arthritis (CIA) rats.

Materials and methods

Forty male Wistar rats were randomly divided into five groups, including model group, normal control group, dexamethasone group, HLJDT group and the mixture of 13 components of HLJDT group after immunized with bovine type II collagen. Urine samples for metabolomic study were collected on 8, 15, 22 day during the animal experiment.

Results

The pharmacological changes (swelling paws and arthritis scores) showed that prophylactic treatment with HLJDT and its components significantly suppressed the swelling of rats’ paws. By combining with partial least squares discriminant analysis, 24 potential biomarkers were identified and primarily involved in 12 metabolism pathways, such as tricarboxylic acids cycle metabolism, lipid metabolism, tryptophan metabolism and phenylalanine metabolism, which revealed a new insight into the RA network in vivo. These potential metabolites identified in CIA model need to be further investigated to prove their diagnostic and/or prognostic values for RA. Taking potential biomarkers found in the study as screening indexes, it revealed that HLJDT and its components could reverse the pathological process of RA through partly regulating the disturbed metabolic pathways.

Conclusions

This study indicated that the metabolomic strategy based on RRLC-TOF-MS is a useful tool to search potential biomarkers related to RA and to dissect the underlying mechanisms of TCM in treating RA.     

Senecio scandens Buch.-Ham.: A review on its ethnopharmacology,phytochemistry, pharmacology,and toxicity

Ethnopharmacological relevance

The aerial parts of Senecio scandens Buch.-Ham. (family Asteraceae) have a long history in traditional Chinese medicine as a treatment for various ailments, such as bacterial diarrhea, enteritis, conjunctivitis, and respiratory tract infections.

Materials and methods

A bibliographic investigation of Senecio scandens was accomplished by analyzing secondary sources, including the Chinese Medicinal plantal Classics, the Internet (Google Scholar and Baidu Scholar), and scientific databases accepted worldwide (Pubmed, Scopus, Web of Science, SciFinder, and CNKI). These sources were scrutinized for available information about the uses of Senecio scandens in traditional Chinese medicine, phytochemistry, pharmacology, and toxicology.

Results

Senecio scandens is a medicinal plant with a climbing woody stem. Phytochemical studies have shown the presence of numerous valuable compounds, such as flavonoids, alkaloids, phenolic acids, terpenes, volatile oils, carotenoids, and trace elements. Among them, PAs are the characteristic constituents, adonifoline is one of the index ingredients of Senecio scandens. Studies in modern pharmacology have demonstrated that extracts and compounds isolated from Senecio scandens show a wide spectrum of pharmacological activities, including anti-inflammatory, antimicrobial, anti-leptospirosis, hepatoprotective, anti-infusorial, antioxidant, antiviral, antitumoral, analgesic, mutagenic, and toxicological activities.

Conclusions

Phytochemical and pharmacological studies have demonstrated that the extracts of the plant possess various pharmacological activities that can be attributed to the presence of various flavonoids, phenolic acids, and alkaloids. Newer technologies for qualitative and quantitative methods of PAs need to be developed to obtain better accuracy and sensitivity. Due to the toxicity of PAs present in this medicinal plant, the regulations on PAs of Senecio scandens were varied among different countries and regions. In China, the PAs toxicity of Senecio scandens ranking criteria is not well defined in the Chinese Pharmacopoeia 2010, and there is no strict uniform requirement on the PAs in western countries. We propose that the use of Senecio scandens should be reevaluated based on a set of criteria, which includes risk–benefit analysis and severity of the toxic effects, clinical and preclinical data to ensure safe use while continuing to satisfy the need for access to the medicinal plant.     

A promising approach for understanding the mechanism of Traditional Chinese Medicine by the aggregation morphology

Objective

Previous work has found that the Traditional Chinese Medicine (TCM) form aggregates in the aqueous solution, and the activities of two Chinese herbal formulae against three cardiovascular targets were aggregates-related. This paper further studied the molecular morphology composed of aggregation and single active molecule in TCM.

Methods

We take Pueraria lobata (Willd.) Ohwi (PUE) as an example. By means of dynamic light scattering (DLS), transmission electron microscopy (TEM), and high performance liquid chromatography–mass spectrometry (HPLC–MS), the mechanism and active components of the aggregates in PUE have been studies. Besides the relationship between aggregation and therapeutic activities in the vivo level has been studied by hemorheological method.

Results

Puerarin, daidzein, daidzin, genistein, these cardiovascular bioactive compounds existed in the aggregates. Three kinds of aggregation processes by the bioactive molecules in the solution were elucidated: (1) the aggregation of single molecule oneself; (2) the aggregation between different single molecules; (3) the aggregation between different single molecules and the primary metabolites. Furthermore, the therapeutic activity of PUE solution was aggregates-related in vivo level.

Conclusions

The aggregation morphology of molecules in TCM might be a promising way to study the mechanism of TCM, even to develop an approach of new nanomedicine of TCM.     

Pharmacokinetics of aconitine as the targeted marker of Fuzi (Aconitum carmichaeli) following single and multiple oral administrations of Fuzi extracts in rat by UPLC/MS/MS

Ethnopharmacological relevance

Fuzi, which is the processed lateral roots of Aconitum Carmichaeli. Debx and is widely distributed over the southwest provinces of China, is recognised for its anti-inflammatory and analgesic effects.

Aim of the study

The pharmacokinetic properties of Fuzi are inadequately understood. Aconitine, the primary highly toxic ingredient of Fuzi, is well known as the target marker of Fuzi. The purpose of the present study is to investigate the pharmacokinetic behaviours of aconitine in vivo following single and multiple administrations of processed Fuzi extracts and to compare the pharmacokinetic characteristics of aconitine after administrations of pure aconitine or Fuzi extracts as well as compare the difference at single dose and multiple doses. The in vitro aconitine protein binding in plasma through equilibrium dialysis was also examined.

Methods

A high performance liquid chromatography (HPLC) method was developed for the determination of aconitine in Fuzi crude extracts and a fast ultra performance liquid chromatography-tandem mass spectrometry (UPLC/MS/MS) was developed to investigate the pharmacokinetic behaviour of aconitine as the targeted marker of Fuzi.

Results

The absolute bioavailability (F %) after the administration of 0.5 mg/kg aconitine and Fuzi extract (0.118 mg/kg aconitine) in rat was 8.24 ± 2.52% and 4.72 ± 2.66%, respectively. Aconitine absorption was very fast at the t_max 30.08 ± 9.73 min for pure aconitine and 58.00 ± 21.68 min for Fuzi extract administration. Aconitine was also eliminated rapidly with a short half-life (i.v., 80.98 ± 6.40 min) and a low rate of protein bounding (23.9–31.9%). No significance was observed on all the pharmacokinetics parameters following the single and multiple doses of pure aconitine (ANOVA, p > 0.05). However, the absorption of aconitine after multiple administrations of Fuzi extract was much faster than that of a single dose (t_max: 58.00 ± 21.68 vs. 20.00 ± 8.66 min, p < 0.05), and the area under the plasma concentration–time curve (AUC) was higher than that of a single dose.

Conclusions

The pharmacokinetic behaviour of processed Fuzi was determined in this paper. The aconitine has low bioavailability. No variation in the pharmacokinetic behaviours of pure aconitine was observed after single and multiple administrations. In contrast, multiple administrations of processed Fuzi extract could result in variations in its pharmacokinetic behaviour in AUC and t_max indicating that multiple dose might increase the bioavailability of aconitine, which may result in its toxicity. In addition, aconitine has a low protein bounding (23.9–31.9%), resulting in its rapid elimination.     

Hypotensive and vasorelaxant effects of the procyanidin fraction from Guazuma ulmifolia bark in normotensive and hypertensive rats

THE AIM OF THE STUDY: was to investigate the in vivo and in vitro cardiovascular activity of a procyanidin fraction (PCF) obtained from acetone extract of Guazuma ulmifolia bark which has traditionally been used as an antihypertensive agent. RESULTS: 10 mg/kg PCF doses orally administered to sugar-fed hypertensive rats decreased both the systolic arterial pressure and the heart rate, whereas the same doses intravenously administered induced arterial hypotension which was attenuated by NG-nitro-L-arginine methylester (L-NAME 31 mg/kg) pretreatment. In these experiments we employed carbachol as a positive control test. The PCF reduced the contraction induced by norepinephrine (1x10(-7) M) in isolated aortic rings of normotensive (IC50=35.3+/-12.4 ng/mL) and sugar-fed hypertensive (IC50=101.3+/-57.2 ng/mL) rats. This relaxant activity was inhibited by either vascular endothelium removal or L-NAME (30 microM) pretreatment, while indomethacin (10 microM) or atropine (10 microM) had no effect. Preliminary analysis of the PCF by HPLC-DAD-MS and FAB+ mass spectrometry allowed the detection of the main components such as the complex of procyanidin oligomers consisting mainly of tetramers and trimers. CONCLUSIONS: Guazuma ulmifolia bark possesses long-lasting antihypertensive and vasorelaxing properties linked to the endothelium related factors, where nitric oxide is involved.     

Anti-diabetic effects of the acetone fraction of Senna singueana stem bark in a type 2 diabetes rat model

Ethnopharmacological relevance

Senna singueana is currently used in the traditional treatment of diabetes mellitus in Nigeria. The present study examined the anti-diabetic activity of the Senna singueana acetone fraction (SSAF) of stem bark in a type 2 diabetes (T2D) rat model.

Materials and methods

Crude ethyl acetate extract of the Senna singueana stem bark was fractionated with various solvents and the acetone fraction was selected for in vivo studies based on the high α-glucosidase and α-amylase inhibitory activities. In the in vivo study, male Sprague-Dawley rats were induced with T2D and treated with the SSAF at 150 and 300 mg/kg body weight. Several T2D-related parameters were measured in the study.

Results

After 4 weeks of intervention, non-fasting blood glucose concentrations were significantly decreased and the glucose tolerance ability was significantly improved in the SSAF treated groups compared to the diabetic control group. Serum insulin concentrations, pancreatic β-cell function (HOMA-β) and liver glycogen were significantly (P<0.05) increased while serum alanine transaminase, alkaline phosphatase and urea were significantly decreased in the SSAF treated diabetic rats compared to the diabetic control group. Though insignificantly (P>0.05), other T2D-induced abnormalities such as food and fluid intake, body weight, serum lipids, serum fructosamine level and peripheral insulin resistance (HOMA-IR) were also partially ameliorated by the SSAF treatment.

Conclusion

Data of this study suggest that orally administered SSAF could ameliorate most of the T2D-induced abnormalities in a T2D model of rats.     

Comparative pharmacokinetics and bioavailability of four alkaloids in different formulations from Corydalis decumbens

Ethnopharmacological relevance

Corydalis decumbens, a Traditional Chinese Medicine listed in Chinese Pharmacopoeia, is clinically used for the treatment of paralytic stroke, headache, rheumatic arthritis and sciatica in China.

Aim of the study

This study was aimed to compare the pharmacokinetics and bioavailability of protopine, tetrahydropalmatine, bicuculline, and egenine in three formulations prepared from the rhizomes of Corydalis decumbens.

Meterials and methods

Alkaloid extract (CDAs-SFE) was prepared from the rhizomes of Corydalis decumbens by supercritical CO₂ fluid extraction; CDAs-SFE/HPβCD (hydroxypropyl-β-cyclodextrin inclusion complex), and CDAs-SFE/HCl (hydrochloride freeze-dried powder) were resulted from CDAs-SFE through complexation with HPβCD and hydrochloride, respectively. An UFLC–MS/MS method was developed for quantitative analysis of protopine, tetrahydropalmatine, bicuculline and egenine simultaneously in rat plasma after oral administration. The differences of pharmacokinetics and bioavailability of the four alkaloids in three formulations were determined by pharmacokinetics analyses.

Results and conclusions

The C_max, AUC and bioavailability of protopine and tetrahydropalamatine (bioactive components) in CDAs-SFE/HCl were significantly higher than in CDAs-SFE and in CDAs-SFE/HPβCD. In contrast, in CDAs-SFE/HPβCD, AUC and bioavailability of tetrahydropalamatine were significantly lower, while those of bicuculline (toxic compound) appeared to be higher than both in CDAs-SFE and in CDAs-SFE/HCl. The results indicated that CDAs-SFE/HCl was the best beneficial formulation among the three formulations for the alkaloid extract prepared from the rhizomes of Corydalis decumbens, in which protopine and tetrahydropalamatine displayed higher bioavailability, but lower for bicuculline.