Pharmacokinetic Analysis of Diosgenin in Rat Plasma by a UPLC-MS/MS Approach.

Diosgenin, a steroidal sapogenin, has attracted attention worldwide owing to its pharmacological properties, including antitumor, cardiovascular protective, hypolipidemic, and anti-inflammatory effects. The current diosgenin analysis methods have the disadvantages of long analysis time and low sensitivity. The aim of the present study was to establish an efficient, sensitive ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) approach for pharmacokinetic analysis of diosgenin amorphous solid dispersion (ASD) using tanshinone IIA as an internal standard (IS). Male Sprague-Dawley rats were orally administered diosgenin ASD, and orbital blood samples were collected for analysis. Protein precipitation was performed with methanol-acetonitrile (50 : 50, v/v), and the analytes were separated under isocratic elution by applying acetonitrile and 0.03% formic acid aqueous solution at a ratio of 80 : 20 as the mobile phase. MS with positive electron spray ionization in multiple reaction monitoring modes was applied to determine diosgenin and IS with m/z 415.2⟶271.2 and m/z 295.2⟶277.1, respectively. This approach showed a low limit of quantification of 0.5 ng/ml for diosgenin and could detect this molecule at a concentration range of 0.5 to 1,500 ng/ml (r = 0.99725). The approach was found to have intra- and inter-day precision values ranging from 1.42% to 6.91% and from 1.25% to 3.68%, respectively. Additionally, the method showed an accuracy of -6.54 to 4.71%. The recoveries of diosgenin and tanshinone IIA were 85.81-100.27% and 98.29%, respectively, with negligible matrix effects. Diosgenin and IS were stable under multiple storage conditions. Pharmacokinetic analysis showed that the C max and AUC0⟶t of diosgenin ASD were significantly higher than those of the bulk drug. A sensitive, simple, UPLC-MS/MS analysis approach was established and used for the pharmacokinetic analysis of diosgenin ASD in rats after oral administration.

Synthesis, in vitro ADME profiling and in vivo pharmacological evaluation of novel glycogen phosphorylase inhibitors.

A small set of indole-2-carboxamide derivatives identified from a high-throughput screening campaign has been described as a novel, potent, and glucose-sensitive inhibitors of glycogen phosphorylase a (GPa). Among this series of compounds, compound 2 exhibited moderate GP inhibitory activity (IC50 = 0.29 µM), good cellular efficacy (IC50 = 3.24 µM for HepG2 cells and IC50 = 7.15 µM for isolated rat hepatocytes), together with good absorption, distribution, metabolism, and elimination (ADME) profiles. The in vivo animal study revealed that compound 2 significantly inhibited an increase of fasting blood glucose level in adrenaline-induced diabetic mice.

Urotensin receptor antagonist urantide improves atherosclerosis-related kidney injury by inhibiting JAK2/STAT3 signaling pathway in rats.

OBJECTIVE: To investigate the role of urantide in the prevention and treatment of atherosclerotic nephropathy by antagonizing the urotensin II/urotensin receptor (UII/UT) system and regulating JAK2/STAT3 signaling pathway. METHODS: Atherosclerosis (AS) rats were treated with urantide at a concentration of 30 µg/kg for 3, 7, 14 days. RESULTS: An excessive expression of UII and its receptor G protein-coupled receptor 14 (GPR14) was seen in AS rat kidneys and the expression was significantly reduced after urantide administration. Either body weight, renal functions of urea nitrogen, urine proteins and anion gaps or expression of kidney injury-related genes Agtr1α, Nox4, Cyba and Ncf1 were improved after AS rats were treated with urantide. After antagonizing the UII/GPR14 system by using urantide, the expression of genes and proteins in the JAK2/STAT3 and ERK pathways was decreased, and the nuclear protein p-STAT3 and p-ERK were obviously decreased. p-JAK2 and p-STAT3 were decreased in the urantide group in a time-dependent manner. The UII/GPR14 system and JAK2/STAT3 signals were localized in tubules and then glomeruli to affect renal reabsorption and filtration. CONCLUSION: Urantide can effectively block the UII/GPR14 system by regulating the JAK2/STAT3 signaling pathway to prevent and treat atherosclerosis-related kidney injury. At this stage, effective inhibition of inflammatory signaling pathways is of great significance in the treatment of atherosclerosis.

Systematic review of long-term Xingnao Kaiqiao needling effcacy in ischemic stroke treatment.

OBJECTIVE: To systematically evaluate the long-term effect and safety of Xingnao Kaiqiao needling method in ischemic stroke treatment. DATA RETRIEVAL: We retrieved relevant random and semi-random controlled trials that used the Xingnao Kaiqiao needling method to treat ischemic stroke compared with various control treatments such as conventional drugs or other acupuncture therapies. Searched databases included China National Knowledge Infrastructure, Weipu Information Resources System, Wanfang Medical Data System, Chinese Biomedical Literature Database, Cochrane Library, and PubMed, from May 2006 to July 2014. SELECTION CRITERIA: Two authors independently conducted literature screening, quality evaluation, and data extraction. The quality of articles was evaluated according to the Cochrane Reviewers' Handbook 5.1, and the study was carried out using Cochrane system assessment methods. RevMan 5.2 was used for meta-analysis of the included studies. MAIN OUTCOME MEASURES: Mortality rate, recurrence rate, and disability rate were observed. RESULTS: Nine randomized and semi-randomized controlled trials treating 931 cases of ischemic stroke were included in this review. Meta-analysis results showed that there were no significant differences in mortality reduction (risk ratio (RR) = 0.58, 95% confidence interval (CI): 0.17-1.93, Z = 0.89, P = 0.37) or recurrence rate (RR = 0.55, 95%CI: 0.18-1.70, Z = 1.04, P = 0.30) of ischemic stroke patients between the Xingnao Kaiqiao needling and control treatment groups. However, the Xingnao Kaiqiao needling method had a tendency towards higher efficacy in mortality reduction and recurrence rates. The Xingnao Kaiqiao needling method was significantly better than that of the control treatment in reducing disability rate (RR = 0.51, 95%CI: 0.27-0.98, Z = 2.03, P < 0.05). CONCLUSION: The Xingnao Kaiqiao needling method has a better effect than control treatment in reducing disability rate. The long-term effect of Xingnao Kaiqiao needling against ischemic stroke is better than that of control treatment. However, the limitations of this study limit the strength of the conclusions. Randomized controlled trials with a strict, reasonable design, and multi-center, large-scale samples and follow-up are necessary to draw conclusions about Xingnao Kaiqiao needling.