Crocin Ameliorates Diabetic Nephropathy through Regulating Metabolism, CYP4A11/PPARγ, and TGF-ß/Smad Pathways in Mice.

INTRODUCTION: Crocin is one of the main components of Crocus sativus L. and can alleviate oxidative stress and inflammation in diabetic nephropathy (DN). However, the specific mechanism by which crocin treats DN still needs to be further elucidated. METHOD: In the present study, a mouse model of DN was first established to investigate the therapeutic effect of crocin on DN mice. Subsequently, non-targeted metabolomics techniques were used to analyze the mechanisms of action of crocin in the treatment of DN. The effects of crocin on CYP4A11/PPARγ and TGF-ß/Smad pathway were also investigated. RESULT: Results showed that crocin exhibited significant therapeutic and anti-inflammatory, and anti-oxidative effects on DN mice. In addition, the non-targeted metabolomics results indicated that crocin treatment affected several metabolites in kidney. These metabolites were mainly associated with biotin metabolism, riboflavin metabolism, and arachidonic acid metabolism. Furthermore, crocin treatment upregulated the decreased levels of CYP4A11 and phosphorylated PPARγ, and reduced the increased levels of TGF-ß1 and phosphorylated Smad2/3 in the kidneys of DN mice. CONCLUSION: In conclusion, our study validated the considerable therapeutic, anti-inflammatory, and antioxidative impacts of crocin on DN mice. The mechanism of crocin treatment may be related to the regulation of biotin riboflavin and arachidonic acid metabolism, the activation of CYP4A11/PPARγ pathway, and the inhibition of TGF-ß/Smad pathway in the kidney.

A new perspective on the inhibition of plant photosynthesis by uranium: decrease of root activity and stomatal closure.

Uranium (U) is difficult to be transported from roots to leaves, but it has been reported to inhabit photosynthesis in leaves, so how does this work? In the present study, the effects of U (0-25 µM) on the development and photosynthesis in V. faba seedlings were studied under hydroponics. The results showed that U significantly inhibited the growth and development of V. faba plants, including decreased biomass, water content, lateral root number and root activity. U also led to a large accumulation of reactive oxygen species (ROS) in the leaves which affects leaf structural traits (e.g., decreased leaf area and chlorophyll a content). When U concentration was 25 µM, the net photosynthetic rate (Pn) and transpiration rate (Tr) were inhibited, which were only 66.53% and 41.89% of the control, respectively. Further analysis showed that the stomatal density of leaves increased with the increase of U concentration, while the stomatal aperture and stomatal conductance (Gs) were on the contrary. The results of chlorophyll fluorescence showed that the non-photochemical quenching coefficient (NPQ) increased and the electron transfer rate (ETR) decreased after U exposure, but fortunately, photosystem II (PSII) suffered little damage overall. In conclusion, the accumulation of U in the roots inhibited the root activity, resulting in water shortage in the plants. To prevent water loss, leaves have to regulated stomatal closure at the cost of weakening photosynthesis. These results provide a new insight into the mechanism by which U affects plant photosynthesis.

Unraveling response mechanism of photosynthetic metabolism and respiratory metabolism to uranium-exposure in Vicia faba.

As a natural radionuclide, uranium (U) has obvious phytotoxicity, the purpose of this study is to unravel the response mechanism of U on photosynthetic and respiratory metabolism in plants. Therefore, 14-day-old Vicia faba seedlings were exposed to 0-25 µM U during 72 h. U effects on growth parameters, physiological parameters of plants, and potential phytotoxicity mechanism were investigated by physiological analysis, and metabolome and transcriptome data. U significantly inhibited photosynthesis and respiration of plants. In metabolome analysis, 53 metabolites related to carbohydrate metabolism were identified (13 up-regulated, 12 down-regulated). In transcriptome analysis, U significantly inhibited the expression of photoreactive electron transport chain (up: 0; down: 31), Calvin cycle (up: 0; down: 12) and photorespiration pathway genes (up: 0; down: 8). U significantly inhibited the expression of cellular energy metabolic pathways genes (e.g., glycolysis, TCA cycle, and oxidative phosphorylation pathways) (up 8, down 18). We concluded that U inhibited the expression of genes involved in the photosynthetic metabolic pathway, which caused the decrease of photosynthetic rate. Meanwhile, U inhibited the expression of the electron transport chain genes in the mitochondrial oxidative phosphorylation pathway, which leads to the abnormal energy supply of cells and the inhibition of root respiration rate.

[Process optimization for extraction and purification of polysaccharides from Cistanche deserticola].

In this study, taking Cistanche deserticola in Xinjiang as the experimental material, the optimal process for extracting polysaccharides from C. deserticola with water extraction was studied by using single factor and orthogonal experiment. Its effects on protein removal and polysaccharides retaining were investigated by using Sevag, enzymatic method or combination of these two methods, so as to determine the optimal method for protein removal from polysaccharides of C. deserticola; the decolorization and purification methods such as macroporous resin of AB-8 and activated Carbon were used to determine the optimal process. The results showed that the extraction rate of polysaccharides from C. deserticola was 18.40% during the optimal process of the water extraction as follows: extraction temperature 75 ℃, extraction time 165 min and solid-liquid ratio 1∶55. The protein removal rate can reach 31.40% and polysaccharide retention rate can reach 96.00% under the optimal protein removal process: temperature 50 ℃, time 2 h, and papain dosage 0.2%. The decolorization rate of activated Carbon and macroporous resin called AB-8 was 80.37% and 86.43%, and the recovery rate of polysaccharides was 77.05% and 91.93%, respectively, suggesting that macroporous resin was more suitable for decoloration. Macroporous resin named AB-8 increased the purity of the polysaccharide crude extract from 67.70% to 84.80% under the following conditions: concentration of the sample 4 g·L~(-1), concentration of the eluent 60% ethanol, and the flow rate 1 mL·min~(-1), showing significant purification effect.

Sinomenine reduces iNOS expression via inhibiting the T-bet IFN-γ pathway in experimental autoimmune encephalomyelitis in rats.

Sinomenine is a bioactive alkaloid isolated from the Chinese medicinal plant Sinomenium acutum. It is widely used as an immunosuppressive drug for treating rheumatic and arthritic diseases. In our previous studies, we found that sinomenine reduced cellular infiltration within the spinal cord and alleviated experimental autoimmune encephalomyelitis (EAE) in rats. In this study, we further investigated the mechanisms of sinomenine treatment in EAE rats. In EAE rats, treatment with sinomenine exerted an anti-inducible NO synthase (anti-iNOS) effect, which is related to the reductions of Th1 cytokine interferon-γ (IFN-γ) and its transcription factor, T-bet, in spinal cords. Moreover, sinomenine treatment of splenocytes stimulated with anti-CD3 antibody and recombinant rat interleukin 12 reduced the expression of T-bet and IFN-γ in vitro and also reduced the capability of supernatants of splenocyte culture to induce iNOS expression by primary astrocytes. However, sinomenine had no direct inhibitory effect on iNOS produced by astrocytes cultured with IFN-γ and tumor necrosis factor α in vitro. In conclusion, the anti-iNOS effect of sinomenine on EAE is mediated via the suppression of T-bet /IFN-γ pathway.

Hesperetin inhibits the maturation and function of monocyte-derived dendritic cells from patients with asthma.

Dendritic cells (DCs) are crucial regulators of allergic diseases. Hesperetin, an important bioactive compound in Chinese traditional medicine, has antioxidant and anti-allergic properties. In this study, we examined whether hesperetin influences surface molecule expression, cytokine production, the capacity to induce T cell proliferation, and the underlying signaling pathway in monocyte-derived DCs from patients with allergic asthma. The results show that hesperetin significantly suppressed Der p 1-induced HLA-DR, CD86 and CD83 expression in DCs. However, the secretion of IL-10 was not affected. Hesperetin-treated DCs exhibited a reduced ability to stimulate autologous CD4+ T cells, accompanied by less Th2 polarization. In addition, the Der p 1-induced phosphorylation of IκBα and the translocation of NF-κB p65 were inhibited in the presence of hesperetin. These novel findings provide insight into the immunopharmacological role of hesperetin in DC-based allergic diseases.

Sinomenine, an antirheumatic alkaloid, ameliorates clinical signs of disease in the Lewis rat model of acute experimental autoimmune encephalolmyelitis.

The therapeutic value of an antirheumatic alkaloid, sinomenine (SIN), was investigated in the acute experimental autoimmune encephalomyelitis (EAE) model of multiple sclerosis (MS). SIN is a bioactive alkaloid derived from the Chinese medicinal plant, Sinomenium acutum REHDER & E. H. WILSON (Family Menispermaceae). Chinese doctors have utilized this plant to treat rheumatic and arthritic diseases for over one thousand years. Experiments in which EAE-induced Lewis rats exhibit an acute monophasic episode of disease demonstrated that SIN is effective in preventing clinical signs of disease. The therapeutic effect on disease activity was observed at preonset administration times and at various doses tested. Consistent with disease activity in vivo, SIN-treated animals have reduced cellular infiltration within the spinal cord along with decreased TNF-alpha and IFN-gamma expression levels. SIN can significantly inhibit proliferation response of splenocytes induced by MBP(68-82). TNF-alpha and IFN-gamma, secreted by splenocytes induced by MBP(68-82) are inhibited by SIN by dose-dependence manner. The mRNA levels of CC chemokines, RANTES, MIP-1alpha and MCP-1, are inhibited in SIN-treated EAE rats. The data in this proof of concept study support the premise that SIN may be a promising new therapeutic intervention in MS.

Immunoassay using probe-labelling immunogold nanoparticles with silver staining enhancement via surface-enhanced Raman scattering.

This paper reports a novel immunoassay based on surface-enhanced Raman scattering (SERS) and immunogold labelling with silver staining enhancement. Immunoreactions between immunogold colloids modified by a Raman-active probe molecule (e.g., 4-mercaptobenzoic acid) and antigens, which were captured by antibody-assembled chips such as silicon or quartz, were detected via SERS signals of Raman-active probe molecule. All the self-assembled steps were subjected to the measurements of ultraviolet-visible (UV-vis) spectra to monitor the formation of a sandwich structure onto a substrate. The immunoassay was performed by a sandwich structure consisting of three layers. The first layer was composed of immobilized antibody molecules of mouse polyclonal antibody against Hepatitis B virus surface antigen (PAb) on a silicon or quartz substrate. The second layer was the complementary Hepatitis B virus surface antigen (Antigen) molecules captured by PAb on the substrate. The third layer was composed of the probe-labelling immunogold nanoparticles, which were modified by mouse monoclonal antibody against Hepatitis B virus surface antigen (MAb) and 4-mercaptobenzoic acid (MBA) as the Raman-active probe on the surface of gold colloids. After silver staining enhancement, the antigen is identified by a SERS spectrum of MBA. A working curve of the intensity of a SERS signal at 1585 cm(-1) due to the [small nu](8a) aromatic ring vibration of MBA versus the concentration of analyte (Antigen) was obtained and the non-optimized detection limit for the Hepatitis B virus surface antigen was found to be as low as 0.5 [micro sign]g mL(-1).