The therapeutic mechanism of Yuye decoction on type 2 diabetes mellitus based on network pharmacology and experimental verification.

ETHNOPHARMACOLOGICAL RELEVANCE: Yuye decoction (YYD) has been widely used as a folk Chinese herbal formula in clinical treatment of type 2 diabetes mellitus(T2DM) for many years. However, its mechanism is still unclear. AIM OF THE STUDY: The aim of this study was to explore the potential mechanism of YYD against T2DM initially by UHPLC-MS/MS combining with network pharmacology, molecular docking techniques and experimental validation. MATERIALS AND METHODS: The main ingredients in the water extract of YYD were initially identified using UHPLC-MS/MS analysis. Combined with network pharmacology and molecular docking techniques, the YYD key compounds-core targets-key signaling pathways network was constructed and the binding activity of key components to core targets was validated. The T2DM rat model was induced by Streptozotocin combined with high glucose and high fat diets. The apoptosis cell model of mouse islet ß-cell of Min6 was induced by high-glucose and palmitic acid. Histopathological and immunofluorescence satining were used to evaluate pancreatic islet ß-cell function and apoptosis in rats. Min6 cell viability and apoptosis ratio were evaluated by CCK-8 and TUNEL staining. The predicted targets and pathways were validated by experiments in vitro and in vivo. RESULTS: The 56 compounds from YYD were identified by UHPLC-MS/MS. The potential targets of the above compounds were predicted by online compound target database, among of which 362 targets were associated with T2DM. Protein-protein interaction analysis identified the main targets such as SRC, MAPK1, PIK3R1, AKT1, HRAS and HSP90AA1, which were considered as the therapeutic targets of YYD on against T2DM. Functional enrichment analysis revealed that PI3K/AKT, FoxO and apoptosis signaling pathways were significantly enriched. Molecular docking results showed that compounds of monolinolein, neomangiferin, mangiferin, pelargonidin-3-O-glucoside and acacetin from YYD had high binding activities to PIK3R1, AKT1, Sirt1 and FoxO1. Therefore, PI3K/AKT1, Sirt1/FoxO1 and apoptotic signaling pathways were considered as predicted targets for experimental validation study. Animal experiments showed that YYD reduced blood glucose levels, improved pancreatic dysfunction and pancreatic islet ß-cells apoptosis in T2DM rats which contributed to the activation of AKT1 and FoxO1 and their related signaling molecules. These results were confirmed in Min6 cell model induced by high-glucose and palmitic acid. CONCLUSIONS: In summary, this study systematically visualized the possible therapeutic effects and mechanisms of YYD on T2DM through the network pharmacology approach and experimental study. The results indicated that YYD could prevent pancreatic islet dysfunction and reverse islet of ß-cells apoptosis possibly via PI3K/AKT1, Sirt1/FoxO1 signaling pathways.

Integrated electro-coagulation and gravity driven ceramic membrane bioreactor for roofing rainwater purification: Flux improvement and extreme operating case.

The collected roofing rainwater with high water quality and large water volume, can alleviate the crisis of water resources and fit the Low-Impact Development (LID) concept. In this work, a novel water purification technology, Electro-Coagulation coupled with Gravity-Driven Ceramic Membrane Bio-Reactor (EC-GDCMBR) was developed for the roofing rainwater purification under long-term operation (136 days). EC-GDCMBR system not only exhibited the better effluent quality, but also obtained the greater flux (~32 LMH). The reason contributed to the high permeability of ceramic membrane and large porosity of biofilm formed by floc growth (~36 µm) during the EC process, which was also proved by SEM image. The coagulation, adsorption, biodegradation, and coprecipitation of EC-GDCMBR was able to synergistically remove the particulate matter, ammonia nitrogen (NH3-N), Total Phosphorus (TP), organic substances, and heavy metal (i.e., Cr, Zn, and Cu). In particular, via the analysis of bacterial abundance, Extracellular Polymeric Substances (EPS), Assimilable Organic Carbon (AOC), Adenosine Tri-Phosphate (ATP) and Confocal Laser Scanning Microscopy (CLSM), EC could sweep most free bacteria on the ceramic membrane surface, enhancing the biological purification efficiency. Furthermore, a large amount of Pseudomonas (12.4 %-66.7 %) and Nitrospira (1.46 %-3.16 %) in the aggregates formed the biofilms, improved the NH3-N removal. During the long-term operation, there are some unavoidable problems, such as the thick and ripened biofilm of EC-GDCMBR would crack and fall off. Based on this, the current work also studied the reliability of GDCMBR under "extreme operating case", and the results showed that neither the biofilm detachment nor the biofilm breakup had a significant impact on the effluent quality. Overall, the findings of this study suggest the reliability of EC-GDCMBR for the sustainable operation of roofing rainwater purification and improve the application value of decentralized rainwater harvest device.

Chronopharmacology of simvastatin on hyperlipidaemia in high-fat diet-fed obese mice.

The chronopharmacology refers to the utilization of physiological circadian rhythms to optimize the administration time of drugs, thus increasing their efficacy and safety, or reducing adverse effects. Simvastatin is one of the most widely prescribed drugs for the treatment of hypercholesterolaemia, hyperlipidemia and coronary artery disease. There are conflicting statements regarding the timing of simvastatin administration, and convincing experimental evidence remains unavailable. Thus, we aimed to examine whether different administration times would influence the efficacy of simvastatin. High-fat diet-fed mice were treated with simvastatin at zeitgeber time 1 (ZT1) or ZT13, respectively, for nine weeks. Simvastatin showed robust anti-hypercholesterolaemia and anti-hyperlipidemia effects on these obese mice, regardless of administration time. However, simvastatin administrated at ZT13, compared to ZT1, was more functional for decreasing serum levels of total cholesterol, triglycerides, non-esterified free fatty acids and LDL cholesterol, as well as improving liver pathological characteristics. In terms of possible mechanisms, we found that simvastatin did not alter the expression of hepatic circadian clock gene in vivo, although it failed to change the period, phase and amplitude of oscillation patterns in Per2::Luc U2OS and Bmal1::Luc U2OS cells in vitro. In contrast, simvastatin regulated the expression of Hmgcr, Mdr1 and Slco2b1 in a circadian manner, which potentially contributed to the chronopharmacological function of the drug. Taken together, we provide solid evidence to suggest that different administration times affect the lipid-lowering effects of simvastatin.

Trace Elements, PPARs, and Metabolic Syndrome.

Metabolic syndrome (MetS) is a constellation of metabolic derangements, including central obesity, insulin resistance, hypertension, glucose intolerance, and dyslipidemia. The pathogenesis of MetS has been intensively studied, and now many factors are recognized to contribute to the development of MetS. Among these, trace elements influence the structure of proteins, enzymes, and complex carbohydrates, and thus an imbalance in trace elements is an independent risk factor for MetS. The molecular link between trace elements and metabolic homeostasis has been established, and peroxisome proliferator-activated receptors (PPARs) have appeared as key regulators bridging these two elements. This is because on one hand, PPARs are actively involved in various metabolic processes, such as abdominal adiposity and insulin sensitivity, and on the other hand, PPARs sensitively respond to changes in trace elements. For example, an iron overload attenuates hepatic mRNA expression of Ppar-α; zinc supplementation is considered to recover the DNA-binding activity of PPAR-α, which is impaired in steatotic mouse liver; selenium administration downregulates mRNA expression of Ppar-γ, thereby improving lipid metabolism and oxidative status in the liver of high-fat diet (HFD)-fed mice. More importantly, PPARs' expression and activity are under the control of the circadian clock and show a robust 24 h rhythmicity, which might be the reasons for the side effects and the clinical limitations of trace elements targeting PPARs. Taken together, understanding the casual relationships among trace elements, PPARs' actions, and the pathogenesis of MetS is of great importance. Further studies are required to explore the chronopharmacological effects of trace elements on the diurnal oscillation of PPARs and the consequent development of MetS.

Silibinin A decreases statin­induced PCSK9 expression in human hepatoblastoma HepG2 cells.

Hypercholesterolemia is one of the major risk factors for the occurrence and development of atherosclerosis. The most common drugs used to treat hypercholesterolemia are 3­hydroxy­3­methyl­glutaryl­CoA reductase inhibitors, known as statins. Statins induce a beneficial increase in the levels of the low density lipoprotein receptor (LDLR) and additionally upregulate proprotein convertase subtilisin/kexin type 9 (PCSK9), which leads to LDLR degradation. This process causes a negative feedback response that attenuates the lipid lowering effects of statins. Therefore, the development of PCSK9 inhibitors may increase the lipid­lowering functions of statins. In the present study, a drug­screening assay was developed using the human PCSK9 promoter, based on data from a dual­luciferase reporter assay, and the efficacies of various compounds from Traditional Chinese Medicine were examined. Among the compounds examined, SIL was demonstrated to function by targeting PCSK9. It was identified that SIL treatment decreased the expression levels of PCSK9 in HepG2 cells by decreasing the activity of the PCSK9 promoter in a dose­and time­dependent manner. Notably, SIL antagonized the statin­induced phosphorylation of the p38 MAPK signaling pathway. The present study suggested that SIL may be developed as a novel PCSK9 inhibitor that may increase the efficiency of statin treatment.

Regulation of reproduction by the circadian rhythms.

Mammals synchronize their circadian activity primarily to the cycles of light and darkness in the environment. Circadian rhythm is controlled by the central clock in the hypothalamic suprachiasmatic nucleus (SCN) and the peripheral clocks in various tissues. More importantly, the central clock can integrate photic/nonphotic signals to generate rhythmic outputs, and then drive the slave oscillators in peripheral tissues through neuroendocrine and behavioral signals. Human reproductive activities, as some other physiological functions, are controlled by the biological clocks. Accumulating lines of epidemiological and genetic evidence indicate that disruption of circadian clock can be directly involved in multiple pathological processes, including infertility. In this review, we mainly discuss the presence of a circadian clock in reproductive tissues and its roles in follicles development, ovulation, spermatogenesis, fertilization and embryo implantation, etc. As the increased shift work and assisted reproductive technologies possibly disrupt circadian rhythmicity to impact reproduction, the importance of circadian rhythms should be highlighted in the regulation of reproductive process.

Graphite oxide-supported CaO catalysts for transesterification of soybean oil with methanol.

Graphite oxide (GO) supported CaO catalysts were prepared and successfully applied to the transesterification of soybean oil with methanol. The supports and resultant catalysts were characterized using X-ray diffraction (XRD), transmission electron microscopy (TEM), N(2) adsorption, thermogravimetry (TG), X-ray photoelectron spectroscopy (XPS), temperature-programed desorption (TPD) and Fourier-transform infrared spectroscopy (FT-IR). The GO supported CaO catalysts exhibited excellent catalytic activity and were easily regenerated by simple heat-treatment. The oxygen-containing groups (i.e., hydroxyl, epoxide groups and carboxyl groups) present on the surface of GO likely act as anchoring centers for CaO. This work demonstrates that graphite oxide is an effective host material of catalytically active CaO nanoparticles for the transesterification of soybean oil with methanol to produce biodiesel.