Effect of Oregon grape root extracts on P-glycoprotein mediated transport in in vitro cell lines.

Purpose: This study aims to investigate the potential of Oregon grape root extracts to modulate the activity of P-glycoprotein. Methods: We performed 3H-CsA or 3H-digoxin transport experiments in the absence or presence of two sources of Oregon grape root extracts (E1 and E2), berberine or berbamine in Caco-2 and MDCKII-MDR1 cells. In addition, real time quantitative polymerase chain reaction (RT-PCR) was performed in Caco-2 and LS-180 cells to investigate the mechanism of modulating P-glycoprotein. Results: Our results showed that in Caco-2 cells, Oregon grape root extracts (E1 and E2) (0.1-1 mg/mL) inhibited the efflux of CsA and digoxin in a dose-dependent manner. However, 0.05 mg/mL E1 significantly increased the absorption of digoxin. Ten µM berberine and 30 µM berbamine significantly reduced the efflux of CsA, while no measurable effect of berberine was observed with digoxin. In the MDCKII-MDR1 cells, 10 µM berberine and 30 µM berbamine inhibited the efflux of CsA and digoxin. Lastly, in real time RT-PCR study, Oregon grape root extract (0.1 mg/mL) up-regulated mRNA levels of human MDR1 in Caco-2 and LS-180 cells at 24 h. Conclusion: Our study showed that Oregon grape root extracts modulated P-glycoprotein, thereby may affect the bioavailability of drugs that are substrates of P-glycoprotein.

Chromosome-level genome of Entada phaseoloides provides insights into genome evolution and biosynthesis of triterpenoid saponins.

As a medicinal herbal plant, Entada phaseoloides has high levels of secondary metabolites, particularly triterpenoid saponins, which are important resources for scientific research and medical applications. However, the lack of a reference genome for this genus has limited research on its evolution and utilization of its medicinal potential. In this study, we report a chromosome-scale genome assembly for E. phaseoloides using Illumina, Nanopore long reads and high-throughput chromosome conformation capture technology. The assembled reference genome is 456.18 Mb (scaffold N50 = 30.9 Mb; contig N50 = 6.34 Mb) with 95.71% of the sequences anchored onto 14 pseudochromosomes. E. phaseoloides was estimated to have diverged from the Leguminosae lineage at ~72.0 million years ago. With the integration of transcriptomic and metabolomic data, gene expression patterns and metabolite profiling of E. phaseoloides were determined in different tissues. The pattern of gene expression and metabolic profile of the kernel were distinct from those of other tissues. Furthermore, the evolution of certain gene families involved in the biosynthesis of triterpenoid saponins and terpenes was analysed and offers new insights into the formation of these two metabolites. Four CYP genes, one UGT gene and related transcription factors were identified as candidate genes contributing to regulation of triterpenoid saponin biosynthesis. As the first high-quality assembled reference genome in the genus Entada, it will not only provide new information for the evolutionary study of this genus and conservation biology of E. phaseoloides but also lay a foundation for the formation and utilization of secondary metabolites in medicinal plants.

LC-Q-TOF/MS-oriented systemic metabolism study of pedunculoside with in vitro and in vivo biotransformation.

As a triterpene saponin, pedunculoside is one of the most abundant, representative and active components in plants of genus Ilex (Aquifoliaceae). Pedunculoside has been used to treat myocardial ischemia, ameliorate hyperlipidemia and prevent liver injury. In this paper, a systemic in vitro liver microsomes / S9 and intestinal bacteria incubation, and in vivo animal experiment were performed, using LC-Q-TOF/MS analysis and a three-step data processing protocol. As a result, Bifidobacterium adolescentis and Bifidobacterium breve were identified to potentially metabolize pedunculoside among the intestinal bacteria tested. A total of 11 metabolites were found and tentatively identified, with 6 in both microsomal and bacterial incubation systems, and 9 after rats orally administered with pedunculoside. The metabolites detected involving both phase I and phase II metabolism, mainly through deglycosylation (hydrolyzation), dehydrogenation, hydroxylation and conjugation, and some of them underwent more than one-step metabolic reactions. Most of the metabolites have not been reported before. In vitro, liver microsome and intestinal bacteria prefer to metabolize pedunculoside in totally different ways; while in vivo, intestinal tract is the most important site for the metabolism and excretion of pedunculoside, where both intestinal bacteria and the host metabolic enzymes participate in its metabolism and disposition. The importance of intestinal bacteria should be highlighted. This study would contribute to a better understanding of pedunculoside metabolism, which can provide scientific evidence for its pharmacodynamic mechanism research and prove its clinical application.

C-Phycocyanin inhibits hepatic gluconeogenesis and increases glycogen synthesis via activating Akt and AMPK in insulin resistance hepatocytes.

C-Phycocyanin (C-PC), a kind of blue protein isolated from Spirulina platensis, can ameliorate hyperglycemia, but its effects on gluconeogenesis and glycogenesis are unknown. In the present study, we investigated the effects and underlying mechanisms of C-PC on gluconeogenesis and glycogenesis in insulin resistant hepatocytes. Insulin resistance was induced by high glucose (HG) in human hepatocellular carcinoma (HepG2) cells. C-PC ameliorated glucose production and phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) expression in HG-induced insulin resistant HepG2 cells. It also increased glucose uptake, glycogen content and glycogen synthase (GS) activation in HG-induced insulin resistant HepG2 cells. The data revealed the mechanism of C-PC in improving glucose homoeostasis via activating the IRS/PI3 K/Akt and SIRT1/LKB1/AMPK signaling pathway in insulin resistant hepatocytes. C-PC could be a promising leading compound for the development of a hypoglycemic agent.

A comprehensive in vitro and in vivo metabolism study of hydroxysafflor yellow A.

As the most important marker component in Carthamus tinctorius L., hydroxysafflor yellow A (HSYA) was widely used in the prevention and treatment of cardiovascular diseases, due to its effect of improving blood supply, suppressing oxidative stress, and protecting against ischemia/reperfusion. In this paper, both an in vitro microsomal incubation and an in vivo animal experiment were conducted, along with an LC-Q-TOF/MS instrument and a 3-step protocol, to further explore the metabolism of HSYA. As a result, a total of 10 metabolites were searched and tentatively identified in plasma, urine, and feces after intravenous administration of HSYA to male rats, although no obvious biotransformation was found in the simulated rat liver microsomal system. The metabolites detected involving both phase I and phase II metabolism including dehydration, deglycosylation, methylation, and glucuronic acid conjugation. A few of the metabolites underwent more than one-step metabolic reactions, and some have not been reported before. The study would contribute to a further understanding of the metabolism of HSYA and provide scientific evidence for its pharmacodynamic mechanism research and clinical use.

Clinical Observation of Chinese Medicinal Recipe Psoriasis No.1 Formula in Treating Blood-heat Type of Psoriasis / 广州中医药大学学报

Objective To observe the clinical efficacy and safety of Chinese medicinal recipe Psoriasis No.1 Formula in treating blood-heat type of psoriasis. Methods Eighty patients with blood-heat syndrome were randomly divided into trial group and control group,40 cases in each group. The trial group was given oral use of the decoction of Psoriasis No . 1 Formula (mainly composed of Radix Rehmanniae, Rhizoma Imperatae, Cortex Moutan,Rhizoma Smilacis Glabrae,Radix Glycyrrhizae,Flos Sophorae,Radix Arnebiae seu Lithospermi, Radix Paeoniae Rubra, Folium Isatidis, Periostracum Cicadae, Radix Scutellariae, and Radix Angelicae Sinensis). The control group was given Compound Qingdai Capsules orally. The treatment for the two groups covered 8 weeks. Psoriasis Area Severity Index(PASI)scores,and contents of interleukin-17(IL-17)and tumor necrosis alpha (TNF-α)were observed before and after treatment. And clinical efficacy and adverse reactions were compared between the two groups after treatment. Results (1) PASI scores of the two groups were significantly lowered after treatment(P < 0.01 compared with those before treatment),and the trial group had stronger effect on decreasing PASI scores than the control group(P<0.05).(2)The total effective rate of the trial group was 87.5％,higher than the control group(67.5％),but the difference was insignificant(P > 0.05). (3)After treatment , contents of IL-17 and TNF-α of the two groups were markedly decreased(P < 0.01 compared with those before treatment),but the difference was insignificant between the two groups(P > 0.05). (4)No severe adverse effect was found in the two groups during the treatment. Conclusion Chinese medicinal recipe Psoriasis No.1 Formula is effective and safe in treating blood-heat type of psoriasis, and its effect is superior to that of Compound Qingdai Capsules.

Key Findings from Preclinical and Clinical Drug Interaction Studies Presented in New Drug and Biological License Applications Approved by the Food and Drug Administration in 2014.

Regulatory approval documents contain valuable information, often not published, to assess the drug-drug interaction (DDI) profile of newly marketed drugs. This analysis aimed to systematically review all drug metabolism, transport, pharmacokinetics, and DDI data available in the new drug applications and biologic license applications approved by the U.S. Food and Drug Administration in 2014, using the University of Washington Drug Interaction Database, and to highlight the significant findings. Among the 30 new drug applications and 11 biologic license applications reviewed, 35 new molecular entities (NMEs) were well characterized with regard to drug metabolism, transport, and/or organ impairment and were fully analyzed in this review. In vitro, a majority of the NMEs were found to be substrates or inhibitors/inducers of at least one drug metabolizing enzyme or transporter. In vivo, when NMEs were considered as victim drugs, 16 NMEs had at least one in vivo DDI study with a clinically significant change in exposure (area under the time-plasma concentration curve or Cmax ratio ≥2 or ≤0.5), with 6 NMEs shown to be sensitive substrates of cytochrome P450 enzymes (area under the time-plasma concentration curve ratio ≥5 when coadministered with potent inhibitors): paritaprevir and naloxegol (CYP3A), eliglustat (CYP2D6), dasabuvir (CYP2C8), and tasimelteon and pirfenidone (CYP1A2). As perpetrators, seven NMEs showed clinically significant inhibition involving both enzymes and transporters, although no clinically significant induction was observed. Physiologically based pharmacokinetic modeling and pharmacogenetics studies were used for six and four NMEs, respectively, to optimize dosing recommendations in special populations and/or multiple impairment situations. In addition, the pharmacokinetic evaluations in patients with hepatic or renal impairment provided useful quantitative information to support drug administration in these fragile populations.

Chronic unpredictable stress impairs endogenous antioxidant defense in rat brain.

Many studies have shown that chronic stress can cause neuronal damage and depression, but this exact mechanism still remains unknown. Neurons are vulnerable to lipid peroxidation-induced damage because the major part of neuronal cell membrane is polyunsaturated fatty acids that are substrate for reactive oxygen species. Since endogenous antioxidant defense systems normally eliminate production of reactive oxygen species, deficient antioxidant defense can cause oxidative stress-induced damage. In the present study, to understand the role of endogenous antioxidant defense in chronic stress-induced neuronal damage, we analyzed lipid peroxidation, total antioxidant capacity, and activities of catalase and glutathione peroxidase in frontal cortex, hippocampus and striatum of rats exposed to chronic unpredictable stress. We found that chronic unpredictable stress for four weeks in rats induced depressive-like behaviors such as anhedonia, despair and decreased exploration. Malondialdehyde, a lipid peroxidation product, is increased, but total antioxidant capacity, glutathione peroxidase activity and catalase activity are decreased in brain of rats exposed to chronic unpredictable stress. Our findings suggest that down regulation of endogenous antioxidant defense induces lipid peroxidation contributing a role to chronic stress and depression.

[Study on quality evaluation of Dafo Longjing tea based on near infrared spectroscopy].

Seven quantitative analysis models for Dafo Longjing tea, including tea color, liquor color, aroma, taste, infused leaf, total points of five factors and total points of six factors, were built by applying near infrared spectroscopy combined with partial least squares (NIRS-PLS), in order to find a objective and scientific method for tea quality evaluation. Results showed that both the calibration samples and the prediction samples of the seven models had acquired a high fitting degree when the number of principal components was below 10, and the value of Rc, RMSEC, Rp and RMSEP were between 90.48%-98.43%, 1.14%-2.09%, 90.00%-96.65%, and 1.52% 2.84%, respectively. Among them, the total points of five factors model had the best prediction performance, and the value of Rp and RMSEP was 96.65% and 1.52%, respectively. Moreover, it was also found that the prediction precision of total points models were higher than that of single factor ones. It seems that the quality evaluation of Dafo Longjing tea could be realized by using NIRS-PLS to some extent.

Ent-kaurane diterpenoids from Rabdosia rubescens and their cytotoxic effects on human cancer cell lines.

Two new ent-kaurane diterpenoids, 16,17-exo-epoxide-oridonin ( 1) and 11,15- O,O-diacetyl-rabdoternins D ( 2), together with thirteen known ones, were isolated from the aerial parts of Rabdosia rubescens. Their structures were established on the basis of high-field 1D and 2D NMR methods supported by HRMS. All diterpenoids were tested for cytotoxicity against human Hep G2, COLO 205, MCF-7, and HL-60 cancer cells. The compounds oridonin ( 3), 14- O-acetyl-oridonin ( 4), 1,14- O,O-diacetyl-oridonin ( 5), rosthorin ( 6), effusanin E ( 7), and ponicidin ( 8), as well as six alpha-methylene gamma-ketone bearing diterpenoids, were modestly active in these assays.

Sesquiterpene lactones from Inula britannica and their cytotoxic and apoptotic effects on human cancer cell lines.

Three new sesquiterpenes (1-3), together with four known sesquiterpene lactones, were isolated from the flowers of Inula britannica var. chinensis. Structures were established on the basis of high-field 1D and 2D NMR methods supported by HRMS. All sesquiterpene lactones were tested for cytotoxicity as well as apoptotic ratio in human COLO 205, HT 29, HL-60, and AGS cancer cells. Compounds 3 and 4, two alpha-methylene gamma-lactone-bearing sesquiterpenes, were modestly active in these assays.

[Prospective, multi-centered, randomized and controlled trial on effect of Shenle Capsule in treating patients with IgA nephropathy of Fei-Pi qi-deficiency syndrome].

OBJECTIVE: To observe the effect and safety of Chinese compound Shenle Capsule (SC) in treating IgA nephropathy patients of Fei-Pi qi-deficiency syndrome (FPQD). METHODS: A prospective, multi-centered, randomized, double-blinded, double-mimetic, controlled trial was conducted in 70 IgA nephropathy patients of FPQD syndrome, who were randomly assigned to two groups treated with SC and fosinopril respectively for 12 weeks. The changes of TCM syndrome score, 24 h urine protein quantity (UP), serum creatinine (SCr), hepatic function before and after treatment and the adverse reaction were observed. RESULTS: Compared with those before treatment, the level of UP and the TCM syndrome scores decreased significantly in both groups (P < 0.05 or P < 0.01), while the level of albumin increased significantly after treatment, and the level of total cholesterol, triglycerid and renal function remained unchanged (P > 0.05). There was no significant difference in all the above parameters between the two groups. According to the curative criterion for Chinese medicine and Western medicine, the total effective rate was 83.3% and 66.7% respectively in the SC treated patients, 82.4% and 58.8% respectively in the fosinopril treated patients, showing insignificant difference between them. In addition, no severe adverse event was found. CONCLUSION: SC treatment showed effects similar to that of fosinopril in reducing proteinuria and improving TCM syndrome in IgA nephropathy patients of FPQD syndrome, and could be well tolerated by patients.

Effects of movement exercise combined with electroacupuncture on expression of Nestin in hippocampus den-tate gyrus after cerebral ischemia-reperfusion / 中华物理医学与康复杂志

Objective To observe the effect of movement exercise combined with electroaeupuneture on the expression of Nestin in the hippocampus dentate gyrus (DG) after cerebral ischemia-repeffusion.Methods Fifty- four Wistar rats were used and randomly divided into a control group (Group A),an exercise training group (Group B),and an exercise training combined with electroacupuncture group (Group C).The middle cerebral arteries (MCA) of all the rats were occluded for 1 h,followed by reperfusion for 7,14 and 21 davs.Immunohistochemistry method was used to detect the expression of Nestin in the hippoeampus dentate gyrus.Results The number of Nes- tin-positive cells peaked in DG in all groups on the 7th day after cerebral isehemia-reperfusion.The number of Nes- tin-positive cells in DG ipsilateral to the ischemia-reperfusion lesion were significantly more than those in the opposite side at various time points (P