[Molecular mechanisms of Gupi Xiaoji decoction inducing apoptosis of human hepatoma HepG2 cells].

Objective: To investigate the molecular mechanisms of Gupi Xiaoji decoction on apoptosis of human hepatoma cells HepG2. Methods: HepG2 cells were divided into 4 groups: control group (Control), blank serum group (Blank), Gupi Xiaoji Yin serum group (GPXJY) and cisplatin group (Positive). Eight duplicate holes were set in each group. After treated with Gupi Xiaoji Decoction-containing serum or cisplatin for 24 hours, the cell viability, the number of viable cells, the state of apoptosis, the cell cycle and the mitochondrial membrane potential were detected, and the level of lipid peroxidation (MDA) and glycolysis rate of the cells were detected. The expressions of apoptotic Bax, Bcl-2, and Caspase-3 proteins, and the contents of triacylglycerol (TG), cholesterol (TC), pyruvate and glucose in the cell supernatant were detected. Results: Compared with the control group, in the GPXJY group, the inhibition rate was increased (P＜0.05), the number of cells was decreased, the number of apoptosis-positive cells was increased (P＜0.01), the number of cells in the G1 phase was increased significantly (P＜0.05), and the cell membrane potential was decreased (P＜0.05,P＜0.01), the glycolytic function was inhibited significantly, the MDA level was increased, the expressions of Bax and Caspase-3 in the GPXJY group were increased, and the expression of Bcl-2 was decreased (P＜0.05, P＜0.01). In cell supernatant, the TC, TG and glucose contents were decreased significantly, and the pyruvate content was increased significantly (P＜0.05,P＜0.01). Conclusion: Gupi Xiaoji Decoction can induce apoptosis of HepG2 cells and may play a role in energy metabolism.

Pharmacokinetics of ginsenoside Rg1 in rat medial prefrontal cortex, hippocampus, and lateral ventricle after subcutaneous administration.

The present study aimed to investigate pharmacokinetics of Rg1 in rat medial prefrontal cortex (mPFC), hippocampus (HIP), and lateral ventricle (LV) after subcutaneous injection. For the first time, intracerebral pharmacokinetics of Rg1 was studied in freely moving rats by microdialysis technique. Rg1 concentrations in dialysates were detected by a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method and were revised using in vivo probe-recovery in HIP and LV. The pharmacokinetic parameters were then determined using non-compartmental models. Since the in vivo recoveries remained stable in HIP and LV during 9 h dialysis, average recoveries were used to revise dialysate concentrations. After dosing, Rg1 was soon detected in brain extracellular fluid (bECF) and cerebrospinal fluid (CSF). The elimination of Rg1 was significantly slower in mPFC than in HIP and LV, and significantly greater AUC was obtained in mPFC than in HIP. Rg1 kinetics in bECF and CSF indicate that Rg1 can go across the blood-brain barrier (BBB) and the blood-cerebrospinal fluid barrier (BCSFB), and then immediately distribute to learning and memory-related regions in brain, which may lead to rapid pharmacological onset. There may be active transport and target-mediated disposition of Rg1 in the CNS, which need to be further clarified.

Comparative bioavailability of ferrous succinate tablet formulations without correction for baseline circadian changes in iron concentration in healthy Chinese male subjects: a single-dose, randomized, 2-period crossover study.

BACKGROUND: Ferrous succinate is used for the treatment of iron deficiency anemia. Determining the bioavailability of iron products is a challenge, because iron is naturally present in the blood and some tissues. Therefore, bioequivalence evaluation of ferrous formulations can be affected by the presence of endogenous iron species. Little information regarding the pharmacokinetics of ferrous supplements is available in the healthy Chinese population. OBJECTIVES: The aim of the study was to assess the comparative bioavailability of 200-mg of a test (ferrous succinate,100 mg × 2, Hunan Huana Pharmaceutical Co., Ltd., Hunan, China) and reference (Sulifei, 100 mg × 2, Nanjing Jinling Pharmaceutical Co., Ltd., Nanjing, China) formulation in healthy Chinese male subjects. The study was conducted to meet Chinese State Food and Drug Administration regulatory requirements for approval of a generic formulation of ferrous succinate. METHODS: This study utilized a single-dose randomized, 2-period, crossover design with alternate assignment of subjects to treatment (a single 200-mg [100 mg × 2]) or reference formulation groups. Both groups underwent a 4-day diet equilibration before 2 days of treatment and, finally, a 4-day washout period for the bioequivalence study. Blood samples were collected at 8:00 am on every diet equilibration day, 0 (baseline), 1, 2, 3, 4, 4.5, 5, 6, 9, 12, 24, and 36 hours after drug administration. Iron concentrations were determined using an inductively coupled plasma mass spectrometry. Subjects in both groups were given a standardized diet, with known amounts of iron and calories. The formulations were assumed to be bioequivalent if the 90% CI ratios for C(max) were within 70% to 143% and AUC(0-last) were within 80% to 125%-criteria established by the Chinese Food and Drug Administration. Tolerability was monitored throughout the study by assessing clinical parameters (vital signs, chemistry laboratory) and subject reports. RESULTS: Twenty healthy Han Chinese male subjects (mean age, 26.8 years [range, 20-39 years]; weight, 61.9 kg [range, 52-72 kg]; body mass index, 21.5 kg/m(2) [range,19.1-23.8 kg/m(2)]; and baseline iron values, 1271 µg/L [range 1113-1429 µg/L]) were enrolled and completed the study. Without baseline correction for endogenous iron compound, the mean C(max) measurements of iron for the test and reference formulations were 2981 [621.1] and 3028 [707.4] µg/L, respectively; AUC(0-last) values were 48,460 [9242] and 48,390 [8420] µg/L/h, respectively; T(max) values were 4.3 [1.6] and 3.7 [1.3] hours. The 90% CIs for the ratios of C(max) and AUC(0-last) were 89.9% to 109.2% and 92.5% to 107.7%. No significant difference was found between groups with regard to pharmacokinetic parameters. Both test and reference formulations were well tolerated, with only 2 (10%) subjects who received the reference formulation complaining of mild heartburn that resolved after approximately 1 hour. Another subject (5%) complained of nausea 10 minutes after the test administration, which resolved within 2 hours. The relative bioavailability of the test-reference preparations was 101.6%. CONCLUSION: In this single-dose crossover study in healthy Chinese male subjects, the test and reference formulations of ferrous succinate 200-mg (100 mg × 2,) tablets met the criteria for assuming bioequivalence as defined by the Chinese Food and Drug Administration. Both formulations were well tolerated. Chinese Clinical Trials registration number: ChiCTR-TRC-11001646.

Preparation, optimization and characteristic of huperzine a loaded nanostructured lipid carriers.

The objective of the present work was to study the preparation, optimization and characteristics of Huperzine A (Hup A), an effective Traditional Chinese Medicine treatment of Alzheimer's disease (AD), loaded nanostructured lipid carriers (NLC). NLC were successfully prepared by a modified method of melt ultrasonication followed by high pressure homogenization using Cetyl Palmitate (CP) as the solid lipid, Miglyol((R))812 as the liquid lipid, Soybean phosphatidylcholine (Spc) and Solutol HS15 as the emulsifiers. The best formulation was optimized with a three-factor, three-level Box-Behnken design. Independent variables studied were the amount of the mixed lipid, the amount of the emulsifier mixture and lipid/drug ratio in the formulation. The dependent variables were the particle size, entrapment efficiency (EE) and drug loading (DL). Properties of NLC such as the morphology, particle size, zeta potential, EE, DL and drug release behavior were investigated, respectively. As a result, the designed nanoparticles showed nearly spherical particles with a mean particle size of 120 nm and -22.93 + or - 0.91 mV. The EE (%) and DL (%) could reach up to 89.18 + or - 0.28% and 1.46 + or - 0.05%, respectively. Differential scanning calorimetry (DSC) of Hup A loaded NLC indicated no tendency of recrystallisation. In vitro release studies showed a burst release at the initial stage and followed by a prolonged release of Hup A from NLC up to 96 h. The results suggest that the presented Hup A loaded NLC system is a potential delivery system for improving drug loading capacity and controlled drug release.

Preparation and characterization of nanostructured lipid carriers loaded traditional Chinese medicine, zedoary turmeric oil.

BACKGROUND: The objective of this work was to study the preparation and characteristics of zedoary turmeric oil (ZTO), a traditional Chinese oily medicine, loaded with nanostructured lipid carriers (NLCs). METHOD: Aqueous dispersions of NLC were successfully prepared by melt-emulsification technique using Crodamol SS as the solid lipid, Miglyol 812N as the liquid oil, and soybean phosphatidylcholine (SbPC) as the emulsifier. Properties of NLC such as the particle size and its distribution, the transmission electron microscope (TEM), drug entrapment efficiency (EE), and drug release behavior were investigated, respectively. The Germacrone blood concentration after intravenous administration of ZTO-NLC was determined and compared with that of ZTO-injection. RESULT: As a result, the drug EEs were improved by adding the liquid lipid into the solid lipid of nanoparticles (SLNs). In vitro drug release experiments indicated that the prepared NLC could enhance the drug release rate over the SLN, and the drug release rate could be adjusted by the liquid lipid content in lipid nanospheres. X-ray and differential scanning calorimetry (DSC) measurements revealed that imperfect crystallization occurred in the inner core of the NLC particles. CONCLUSION: The results suggest that the presented NLC system might be a promising intravenous dosage form of water-insoluble oily drugs.