Integrating Network Pharmacology and Experimental Validation to Reveal the Mechanism of Vine Grape Tea Polyphenols on Hyperuricemia-Induced Renal Injury in Mice.

Hyperuricemia (HUA) is a metabolic disease and contributes to renal injury (RI). Vine grape tea polyphenols (VGTP) have been widely used to treat HUA and RI. However, the potential mechanism of VGTP activity remains unclear. To explore the underlying mechanism of VGTP treatment for HUA-induced RI based on network pharmacology that is confirmed by an in vivo study. All ingredients of VGTP were retrieved using a Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and Comparative Toxicogenomics Database systems. The related targets of HUA and RI were obtained from GeneCards and National Center for Biotechnology Information (NCBI) databases. Some ingredients and targets were selected for molecular docking verification. One hour after administering potassium oxonate (300 mg/kg), VGTP (50, 100, and 200 mg/kg/d) was orally administered to HUA mice for 4 weeks. Histopathology and western blotting were performed in renal tissue. Our results showed that VGTP significantly reduced blood urea nitrogen, creatinine, uric acid, and significantly improved the RI and fibrosis of HUA mice. There were 54 active ingredients and 62 targets of HUA-induced RI. Further studies showed that VGTP decreased the expression of Bax, cleaved caspase 3, transforming growth factor-ß (TGF-ß1), CHOP, p-STAT3, and P53, and increased Bcl-2 expression in renal tissue. The related signaling pathways have apoptosis, TGF-ß1, P53 and STAT, and endoplasmic reticulum stress (ERS). In this study, VGTP exerted antihyperuricemic and anti fibrosis effects by regulating the apoptosis and ERS signaling pathways. VGTP is expected to become a drug for combating HUA and RI.

l-Arginine treatment maintains postharvest quality in blueberry fruit by enhancing antioxidant capacity during storage.

The postharvest quality of blueberry fruit is largely limited by deterioration. l-arginine (Arg) is a functional nontoxic amino acid with high biological activities. This study investigated the positive effects and the underlying mechanism of Arg treatment on the quality of postharvest blueberries. Arg effectively mitigated fruit decay and improved the quality of blueberries, including weight loss, firmness, and soluble solid content. Mechanistically, Arg-mediated activation of the anti-oxidative defense system reduced reactive oxygen species-mediated oxidative damage. Moreover, Arg treatment decreased the activities and gene expression of phospholipase D, lipoxygenase, and lipase-inhibiting membrane lipid peroxidation during the prolonged storage of blueberries. Meanwhile, Arg treatment increased nitric oxide (NO) content and NO synthase activity. Furthermore, correlation and principal component analyses revealed the enhancement of Arg treatment on antioxidant capacity. This study suggests that Arg treatment can maintain the postharvest quality of blueberries by improving antioxidant capacity.

High Purity Struvite Recovery from Hydrothermally-Treated Sludge Supernatant Using Magnetic Zirconia Adsorbent.

Recovery of phosphorus from sludge will help to alleviate the phosphorus resource crisis. However, the release of phosphorus from sludge is accompanied by the leaching of large amounts of coexisting ions, i.e., Fe, Al, Ca, and organic matter, which decreases the purity of sludge-derived products. In this study, an adsorption-desorption process using magnetic zirconia (MZ) as the adsorbent is proposed to obtain a high purity recovery product. The process involves selective adsorption of phosphate from the hydrothermally treated sludge supernatant (HTSS) using MZ, followed by desorption and precipitation to obtain the final product: struvite. The results indicated that at a dosage of 15 g/L, more than 95% of phosphorus in the HTSS could be adsorbed by MZ. Coexisting ions (Ca2+, Mg2+, Fe3+, Al3+, SO42-, NO3-, Cl-, etc.) and organic matter (substances similar to fulvic and humic acid) in the HTSS had a limited inhibitory effect on phosphate adsorption. Using a binary desorption agent (0.1 mol/L NaOH + 1 mol/L NaCl), 90% of the adsorbed phosphorus could be desorbed. Though adsorption-desorption treatment, struvite purity of the precipitated product increased from 41.3% to 91.2%. Additionally, MZ showed good reusability, maintaining a >75% capacity after five cycles. X-ray photoelectron spectroscopy (XPS) indicated that MZ adsorbed phosphate mainly by inner-sphere complexation. This study provided a feasible approach for the recovery of phosphorus from sludge with high purity.

Case Report: A Case of Late-Onset Combined Methylmalonic Acidemia and Hyperhomocysteinemia Induced by a Vegetarian Diet.

Methylmalonic acidemia is a rare autosomal recessive metabolic disease. However, because of the atypical clinical symptoms, the type of late-onset methylmalonic academia is often misdiagnosed. Especially when the blood vitamin B12 and folic acid levels are normal, it is not easy to think of this disease. Herein we report a 9-year-old girl who developed normally on a relatively balanced diet before 7 years of age. However, she presented with fatigue and attention deficit when she followed a vegetarian diet. Laboratory examination showed moderate macrocytic anemia, high levels of homocysteine, high level of propionylcarnitine/acetylcarnitine, urinary methylmalonic acid and methyl citrate. Gene mutation analysis showed c.609G > A and c.80A > G compound heterozygous mutations in the MMACHC gene, supported late-onset combined methylmalonic academia with homocysteinemia. Then treatment performed with add meat to the diet, vitamin B12, folic acid betaine and L-carnitine supplement. One week later, the child's clinical symptoms and the laboratory examinations were significantly improved.

Flavonoid biosynthesis in four Dendrobium species based on transcriptome sequencing and metabolite analysis.

BACKGROUND: Dendrobium is a genus of plants used as traditional Chinese herbal medicines, with high economic and medicinal value. METHODS AND RESULTS: To reveal the mechanism of flavonoid biosynthesis in Dendrobium, the metabolites and transcriptomes of four Dendrobium species (D. chrysotoxum, D. nobile, D. fimbriatum, and D. denneanum) were analyzed comprehensively. Ultra-high-performance liquid chromatography-tandem mass spectrometry analysis revealed ten flavonoid compounds in Dendrobium. In total, 100,096 unigenes were obtained from the transcript database of the four Dendrobium species. Among the identified differentially expressed genes, 51 were associated with flavonoid biosynthesis, and 670 differentially expressed transcription factors were predicted, including 194 MYB, 87 bHLH, and 100 WRKY family transcription factors, respectively. Transcriptome analysis showed that the expression levels of structural genes such as chalcone synthase (CHS), cinnamate-4-hydroxylase (C4H), and flavonoid 3'-hydroxylase (F3'H) were lower in D. chrysotoxum, D. nobile, and D. fimbriatum than those in D. denneanum, which may be the main reason for the low flavonoid contents in D. chrysotoxum, D. nobile, and D. fimbriatum. CONCLUSIONS: The expression level of structural genes corresponded to the accumulation level of flavonols in the different Dendrobium species. The results deepen the understanding of the molecular mechanism of flavonoid biosynthesis in Dendrobium and provide novel insights into the synthesis and accumulation of flavonoids in Dendrobium.

The Light-Induced WD40-Repeat Transcription Factor DcTTG1 Regulates Anthocyanin Biosynthesis in Dendrobium candidum.

Dendrobium candidum is used as a traditional Chinese medicine and as a raw material in functional foods. D. candidum stems are green or red, and red stems are richer in anthocyanins. Light is an important environmental factor that induces anthocyanin accumulation in D. candidum. However, the underlying molecular mechanisms have not been fully unraveled. In this study, we exposed D. candidum seedlings to two different light intensities and found that strong light increased the anthocyanin content and the expression of genes involved in anthocyanin biosynthesis. Through transcriptome profiling and expression analysis, we identified a WD40-repeat transcription factor, DcTTG1, whose expression is induced by light. Yeast one-hybrid assays showed that DcTTG1 binds to the promoters of DcCHS2, DcCHI, DcF3H, and DcF3'H, and a transient GUS activity assay indicated that DcTTG1 can induce their expression. In addition, DcTTG1 complemented the anthocyanin deficiency phenotype of the Arabidopsis thaliana ttg1-13 mutant. Collectively, our results suggest that light promotes anthocyanin accumulation in D. candidum seedlings via the upregulation of DcTTG1, which induces anthocyanin synthesis-related gene expression.

Restoration of Mimecan Expression by Grape Seed Procyanidin B2 Through Regulation of Nuclear Factor-kappaB in Mice With Diabetic Nephropathy.

Grape seed procyanidin B2 (GSPB2) exerts a variety of potent protective pharmacological effects on diabetic complications. The renal protective effects of GSPB2 and the target protein mimecan regulated by GSPB2, discovered in a previous quantitative proteomic analysis, were assessed in mice with diabetic nephropathy Twenty-four db/db mice were divided into 2 groups of the vehicle-treated and GSPB2-treated (30 mg/kg/d) diabetic groups. All animals were observed for 10 weeks. Treatment with GSPB2 resulted in an improvement in body weight increase and serum levels of triglyceride, total cholesterol, advanced glycation end products, and urinary albumin excretion in comparison with the vehicle-treated diabetic mice (P < .05), although these levels were still higher than those in the control group. Treatment with GSPB2 significantly reduced the extent of glomerular basement membranes thickening, mesangial expansion, and glomerular area as well. Mimecan protein expressions in diabetes mellitus were decreased approximately by 28% when compared with those in the control group (P < .05), and restored remarkably after GSPB2 treatment (P < .05). The expression of nuclear factor-κB (NF-κB) p65 in nuclear extracts, markedly higher in the diabetic mice than in the controls, was significantly suppressed by GSPB2. The findings of this study revealed that mimecan might become a new therapeutic target in the future and indicated that GSPB2 had beneficial effects not only on oxidative stress, but also on renal fibrosis, particularly in the diabetic kidney.

Ligustrazine improves blood circulation by suppressing Platelet activation in a rat model of allergic asthma.

Chuan-xiong (Ligusticum wallichii) is a traditional herbal medicine in Eastern Asia, but the effect of its active component ligustrazine remains unclear. We explored its effect and possible mechanism in a well-characterized rat model of allergic asthma. Ligustrazine suppressed bronchial hyper-responsiveness to methacholine, and suppressed lung inflammation in asthmatic rats. Ligustrazine exhibited potent immuno-modulatory and anti-inflammatory effects: it suppressed lymphocyte and eosinophil mobilization, and reduced cytokine IL-5 and IL-13 production significantly in lung tissues from asthmatic rats (P<0.05). Further histological examinations clearly demonstrated that ligustrazine improved blood circulation and ameliorated platelet activation, aggregation and adhesion, which induced sustained infiltration and activation of various inflammatory cells, including lymphocytes and eosinophils, followed by synthesis and release of a variety of pro-inflammatory mediators and cytokines. The present study suggests that ligustrazine is a potent agent for the treatment of allergic asthma due to its strong anti-inflammatory and immuno-modulatory properties.

Protective effects of Sapindus saponins in spontaneously hypertensive rats.

OBJECTIVES: To investigate the protective effects of Sapindus saponins in spontaneously hypertensive rats, and the possible cellular and molecular mechanisms. METHODS: Thirty-two 16-week-old spontaneously hypertensive rats were randomly divided into four groups (8 in each group): model group (placebo), positive control group (27 mg/kg of Captopril Tablets), Sapindus saponins groups (27 mg/kg and 108 mg/kg, respectively). Another 8 healthy Wistar-Kyoto strain (WKY) rats were used as the normal group. The animals were treated for 8 weeks. Blood pressure of rats was determined by non-invasive blood pressure meter (BP-6). Furthermore, the contents of angiotensin II (Ang II) in plasma and myocardial tissue were determined by enzyme-linked immunosorbent assay (ELISA), the gene expression of receptor angiotensin type 1 (AT1R) in aorta was determined by quantitative realtime polymerase chain reaction (qRT-PCR). The protein expression of transforming growth factor-ß1 (TGF-ß1) and AT1R in heart was determined by immunohistochemical staining. The protein expression of p-phosphorylation of p38 mitogen-activated protein kinase (p-p38MAPK) was determined by Western blotting. The contents of interleukin (IL)-1, IL-6 and tumor necrosis factor (TNF) in serum were determined by radioimmunoassay. And the histopathological and morphological changes of aorta and heart tissue samples were assessed semi-quantitatively by hematoxylin-eosin (HE) or Masson staining. RESULTS: Thirty minutes after single or continuous treatment, systolic blood pressure (SBP) was reduced significantly in Sapindus saponins groups. And the contents of AngII, IL-1, IL-6 and TNF-α in serum, the expression of AT1R mRNA, p-p38MAPK and TGF-ß1 were significantly suppressed dose-dependently (P<0.05 or P<0.01). With the Sapindus saponins treatment, compared with those of the model group, the cardiac and aortic pathological changes were ameliorated significantly. CONCLUSIONS: Our findings suggest that Sapindus saponins might have protective effects in spontaneously hypertensive rats, the cellular and molecular mechanisms of which might be relevant to the regulation of inflammatory responses mediated by p-p38MAPK signal pathway based on activated Ang II and AT1R.

Delisheng Injection (), a Chinese medicinal compound, enhanced the effect of cis-platinum on lung carcinoma cell line PGCL3.

OBJECTIVE: To investigate the effect of Delisheng Injection (, DLS), a Chinese medicinal compound, DLS combined with cis-platinum (DDP), an active agent used in lung cancer chemotherapy, on a human highly metastatic giant lung carcinoma cell line PGCL3. METHODS: The suspended PGCL3 cells at 10(5) /mL cultured in 96-well tissue culture plates were divided into 4 groups: DLS treatment group (2 µL/mL, 5 µL/mL, 10 µL/mL, 25 µL/mL), DDP treatment group (1 µg/mL, 2 µg/mL, 5 µg/mL, 15 µg/mL), combined DLS with DDP treatment group (DLS:DDP 2 µL/mL:1 µg/mL, 5 µL/mL:2 µg/mL, 10 µL/mL:5 µg/mL, 25 µL/mL:15 µg/mL) and a control group. The cytotoxicity of DLS with different concentrations (2 µL/mL, 5 µL/mL, 10 µL/mL, 25 µL/mL) on PGCL3 cells was determined by 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay. Effect of DLS on adhesion of PGCL-3 cells was tested by cell-matrigel adhesion assay. Chemotactic movement model of transwell camerula was used to determine the effect of DLS on invasion and migration of PGCL-3 cells. RESULTS: Compared with the control group, DLS (2 µL/mL, 5 µL/mL, 10 µL/mL, 25 µL/mL) could significantly decrease cell proliferation, adhesion, invasion and migration abilities (P <0.05). Cell adhesion, invasion and migration abilities were significantly decreased after combination treatment of DLS:DDP (2 µL/mL:1 µg/mL, 5 µL/mL:2 µg/mL, 10 µL/mL:5 µg/mL, 25 µL/mL:15 µg/mL) compared with DDP single-agent treatment (1 µg/mL, 2 µg/mL, 5 µg/mL, 15 µg/mL, P<0.05), respectively. CONCLUSIONS: DLS single-agent has a satisfying inhibition effect in PGCL3 cell line and DLS might enhance the inhibition effect of DDP on cancer metastasis. Our research provided a experimental basis about the treatment on highly metastatic lung caner.

[Effect of Sapindus saponins on myocardial inflammation and left ventricular remodeling in spontaneously hypertensive rats].

OBJECTIVE: To investigate the effect of Sapindus saponins on myocardial inflammation and left ventricular remodeling in spontaneously hypertensive rats. METHODS: Forty 16-week-old spontaneously hypertensive rats were randomly divided into five groups, placebo as model group, captopril tablets (27 mg/kg) as positive control, low-dose Sapindus saponins (27 mg/kg), medium-dose (54 mg/ kg) and high-dose (108 mg/kg) groups. And another eight healthy Wistar-Kyoto strain (WKY) rats were used as the normal group. The animals were treated for eight weeks, and the detection indexes were as follows: (1) Calculated left ventricular mass index (LVMI); (2) Observed the morphological changes on left ventricular myocardial tissue by HE staining; (3) Observed the collagen distribution in left ventricular myocardial by Masson staining; (4) Detected the protein expression of TGF-beta1 by immunohistochemical assay. RESULTS: Sapindus saponins could effectively reverse the left ventricular hypertrophy phenomenon in SHR, lowered LVMI, inhibited the myocardial cell hypertrophy and hyperplasia of collagen fibers, and blocked the expression level of TGF-beta1 in myocardial when compared with the SHR model group, there were significant differences (P < 0.05 or P < 0.01). CONCLUSION: Sapindus saponins can reserve the left ventricular remodeling in pathological conditions, its possible mechanism may be related to the inhibition of myocardial tissue inflammation factor of TGF-beta1.

[Effects of sapindus saponins on inflammatory response mediated by Ang II/p38MAPK pathway and cardiac hypertrophy in spontaneously hypertensive rats].

OBJECTIVE: To investigate the effects of sapindus saponins on myocardial inflammation mediated by Ang II/ p38MAPK signal pathway and cardiac hypertrophy in spontaneously hypertensive rats. And also to explore the correlation of cardiac hypertrophy and inflammation. METHOD: Thirty-two 16-week-old spontaneously hypertensive rats (SHR) were randomly divided into four groups, one with placebo as model group, one with captopril tablets (27 mg x kg(-1)) as positive control, one with low-dose sapindus saponins (27 mg x kg(-1)), one with high-dose (108 mg x kg(-1)). And another eight healthy Wistar-Kyoto strain (WKY) rats were used as the normal group. The animals were treated for eight weeks, and the indicators detected were as follows: (1) left ventricular mass index (LVMI); (2) the content of Ang II and hs-CRP in plasma were determined by ELISA; (3) the protein expression of AT1R and VEGF were determined by immunohistochemical method; (4) the protein expression of p-p38MAPK in myocardial cells was determined by Western blot. RESULT: Sapindus saponins reduced LVMI, and blocked the expression level of Ang II, AT1R, p-p38MAPK, VEGF and hs-CRP in myocardial tissue. Vs the SHR model group, there were significant differences (P < 0.05 or P < 0.01). CONCLUSION: Our findings suggested that sapindus saponins could inhibited cardiac hypertrophy, the possible mechanisms may be related to the inhibition on inflammatory response mediated by Ang II/p38MAPK pathway.

[Research of sapindus saponins on endothelial function in spontaneously hypertensive rats].

OBJECTIVE: To investigate the regulation on endothelial function of sapindus saponins in spontaneously hypertensive rats by studying the reactivity on different vasoconstrictor and dilator, and the content of the active substances. METHOD: Forty 16-week-old spontaneously hypertensive rats were randomly divided into five groups, one with placebo as model group, one with captopril tablets (27 mg x kg(-1)) as positive control, one with low-dose sapindus saponins (27 mg x kg(-1)), one with medium-dose (54 mg x kg(-1)), one with high-dose (108 mg x kg(-1)). And another eight healthy Wistar-Kyoto strain (WKY) rats were used as the normal group. The animals were treated for eight weeks, and the indicators to be detected were as follows: (1) the response of thoracic aorta on different vasoconstrictors Ang II (1 x 10(-9) -1 x 10(-5) mol x L(-1)), PE (1 x 10(-8) 1 x 10(-4) mol x L(-1)), KCl (20 -120 mmol x L(-1)); (2) the endothelium-dependent or non-endothelium-dependent vasodilation response of thoracic aorta on Ach (1 x 10-(10)-1 x 10(-5) mol x L(-1)) or SNP (1 x 10(-8)-1 x 10(-3) mol x (L(-1); (3) the content of NO, 6-KPG1alpha, ET-1 and TXB2 in serum was determined by Elisa. RESULT: In SHR model group, the response of thoracic aorta on Ang II, PE and KCl was increased, the endothelium-dependent vasodilation on Ach was reduced, but the effects on SNP was not obvious, the content of ET-1 and TXB2 was increased, and the content of NO and 6-KPG1alpha was reduced, Vs the normal control group, there were significant differences (P < 0.05 or P < 0.01); in the treatment groups, the response of thoracic aorta on Ang II, PE and KCl was reduced, the endothelium-dependent vasodilation of thoracic aorta on Ach was improved, the content of ET-1 and TXB2 was reduced, and the content of NO and 6-KPG1alpha was increased, Vs the SHR model group, there were significant differences (P < 0.05 or P < 0.01). CONCLUSION: Our findings suggested that sapindus saponins protected the endothelial function in SHR, the mechanisms were relevant to the protection of endothelial function.

Action of a Novel PDE4 inhibitor ZL-n-91 on lipopolysaccharide-induced acute lung injury.

In the present study, we investigated the effect of classic PDE4 inhibitor rolipram and novel PDE4 inhibitor ZL-n-91 on LPS-induced acute lung injury (ALI) in mice and its mechanism. ALI was induced in ICR mice by instilling intratracheally with LPS, and mice were divided into seven groups: control (Saline), LPS group, ZL-n-91 (3 microg, 10 microg, and 30 microg kg(-1), ip), Rolipram (1.0 mg kg(-1), ip) and dexamethasone (0.5 mg kg(-1), ip). After the 6h of instilling intratracheally with LPS in mice, total leukocyte number, neutrophil number and protein content in BALF increased rapidly, a large number of neutrophil infiltration around the pulmonary vessel and airway, the lung wet weight/dry weight (w/d)ratio raised significantly. MPO activity, TNF-alpha level and cAMP-PDE, PDE4 activity in lung homogenate raised significantly. P(a)O(2), P(a)CO(2) and PH value in peripheral arterial blood also changed obviously, P(a)O(2) and PH value dropped slightly and P(a)CO(2) increased significantly in LPS group. ZL-n-91 (3 microg, 10 microg, 30 microg kg(-1)) dose-dependently reduced the total leukocyte number, neutrophil number and total protein content in BALF, MPO activity, TNF-alpha level and cAMP-PDE, PDE4 activity in lung homogenate, but the effect of ZL-n-91 in pathological changes and lung wet w/d ratio is slight; Rol and Dex significantly reduced lung wet w/d ratio and improved pathological changes, neutrophil around the pulmonary vessel and airway significantly reduced, symptoms of lung edema relieved; The PH value, P(a)O(2) and P(a)CO(2) in ZL-n-91 high dosage group and Rol group had changes, but there was no significant difference compared with LPS group or saline group; After the administration, the righting reflex recovery time significantly shorten in every group of ZL-n-91. the righting reflex recovery time of Rol group was similar with ZL-n-91 30 microg kg(-1) group, while Dex group was similar with saline group. The present study confirms that the inhibitory effect of ZL-n-91(30 microg kg(-1)) on the inflammatory reactivity, including inhibition of inflammatory cell and protein exudation, MPO and PDE4 activity, improvement of the blood gas, those effects were equivalent with rolipram 1 mg kg(-1), and suggested that ZL-n-91 was stronger than rolipram in PDE4 inhibition. So we speculated that ZL-n-91 may have stronger therapeutic potential for treatment of inflammatory disease than rolipram, meantime have stronger nervous system effect than rolipram.

[Protective effect of crocetin on primary culture of cardiac myocyte treated with noradrenaline in vitro].

AIM: To investigate the cardio-protective effect of crocetin on primary culture of cardiac myocyte treated with noradrenaline. METHODS: After adding crocetin, the primary culture of cardiac myocyte was injured by 1.0 micromol x L(-1) noradrenaline. The activity of lactic dehydrogenase (LDH), mitochondrion succinic dehydrogenase (MSDH) and ATPase were assayed. The mitochondrion membrane potential was detected by Rh123. The percentage of cardiac myocyte apoptosis was observed by flow cytometry. RESULTS: Crocetin significantly decreased the activity of LDH in culture supernatant, increased the activity of MSDH, ATPase (Na+-K+ ATPase, Ca2+ ATPase) and mitochondrion membrane potential. CONCLUSION: Crocetin could alleviate the disturbance of energy metabolism and decrease the percentage of apoptosis of cardiac myocyte treated with noradrenaline.