RES Treatment Research Articles

Nullifying epigenetic writer DOT1L attenuates neointimal hyperplasia

Background and aimsHistone methyltransferases are emerging targets for epigenetic therapy. DOT1L (disruptor of telomeric silencing 1-like) is the only known methylation writer at histone 3 lysine 79 (H3K79). It is little explored for intervention of cardiovascular disease. We investigated the role of DOT1L in neointimal hyperplasia (IH), a basic etiology of occlusive vascular diseases. Methods and resultsIH was induced via balloon angioplasty in rat carotid arteries. DOT1L and its catalytic products H3K79me2 and H3K79me3 (immunostaining) increased by 4.69 ± 0.34, 2.38 ± 0.052, and 3.07 ± 0.27 fold, respectively, in injured (versus uninjured) carotid arteries at post-injury day 7. Dot1l silencing via shRNA-lentivirus infusion in injured arteries reduced DOT1L, H3K79me2, and IH at day 14 by 54.5%, 37.1%, and 76.5%, respectively. Moreover, perivascular administration of a DOT1L-selective inhibitor (EPZ5676) reduced H3K79me2, H3K79me3, and IH by 56.1%, 58.6%, and 39.9%, respectively. In addition, Dot1l silencing and its inhibition (with EPZ5676) in vivo in injured arteries boosted smooth muscle α-actin immunostaining; pretreatment of smooth muscle cells with EPZ5676 in vitro reduced pro-proliferative marker proteins, including proliferating cell nuclear antigen (PCNA) and cyclin-D1. ConclusionsWhile DOT1L is upregulated in angioplasty-injured rat carotid arteries, either its genetic silencing or pharmacological inhibition diminishes injury-induced IH. As such, this study presents a strong rationale for continued mechanistic and translational investigation into DOT1L targeting for treatment of (re)stenotic vascular conditions.

Mulching practices alter soil microbial functional diversity and benefit to soil quality in orchards on the Loess Plateau

To improve our understanding about the responses of microbial functional diversity to different mulching practices, this study used a metagenomic approach to reveal soil microbial functional specificity under four tillage regimes: conventional tillage (CT), organic mulch practices with ryegrass (Lolium perenne L.) intercropping cover (RE) and cornstalk mulch (CS), and inorganic mulching with black ground fabric (BF) in a 7-year field experiment in an apple orchard of the Loess Plateau in China. Enzyme activity and soil physicochemical properties were measured. A redundancy analysis showed that the RE and CS treatments had positive effects on soil nutrient and enzyme activity compared to that of the BF and CT treatments. The CS and RE treatments increased β-glucosidase, cellobiohydrolase, and β-xylosidase activities. In addition, the CS treatment significantly enhanced the β-N-acetylglucosaminidase and urease activities compared to that under CT treatment. However, the activity of these enzyme was reduced in the BF treatment compared with that of the CT treatment. The results also indicated that the enzymes activities were not completely consistent with the changing trends of the genes encoding these enzymes. In addition, the RE and CS treatments also increased the abundance of genes encoding carbohydrate enzymes. It is interesting that the RE and CS treatments had more pathway genes associated with the carbon cycle, nitrogen cycle, and amino acid metabolism compared with the BF treatment. Remarkably, RE and CS treatments effectively increased the abundance of carbon fixation gene cbbL compared to CT treatment. In summary, organic mulching practices increased the soil microbiological functional diversity related to the carbon and nitrogen cycle, while inorganic mulching practice reduced them. This study enhanced our understanding of how mulching practices may alter soil microbial functional diversity and benefit soil quality.

Effects of feedbunk restrictions and push-up frequency on the growth performance of Holstein dairy heifers offered a forage-based diet with a limit-feeding strategy

Conceptually, there are 2 feeding strategies for avoiding over-conditioning, which can be problematic for gravid dairy heifers that have reduced dietary energy density requirements relative to younger animals: (1) diluting the ad libitum-fed diet with low-energy forages; or (2) offering a diet of greater nutrient density but intentionally restricting the DM available for consumption (limit-feeding). Our objectives for this study were to evaluate the effects of feedbunk restriction and feed push-up frequency on the growth performance of gravid Holstein dairy heifers. A total of 128 Holstein heifers (434 ± 46.7 kg) were enrolled in the trial. Heifers were blocked by weight, and assigned to 1 of 16 identical research pens (4 pens/weight block; 8 heifers/pen), where the mean initial body weight (BW; ± SD) for the 4 blocks were 491 ± 19.0, 450 ± 16.5, 419 ± 10.6, 374 ± 23.0 kg. Within each block, a 2 × 2 factorial arrangement of treatments was assigned; treatments consisted of feedbunk access [full (FUL) or restricted (RES] and feed push-up frequency [1.5- or 3.0-h intervals]. The RES treatment was applied by covering 2 of the 8 head-locking feed gates in assigned pens with plywood partitions, thereby creating a feedbunk-stocking rate of 133%. A total mixed ration diet composed of alfalfa haylage (60.5%), corn silage (38.0%), and mineral (1.5%) was offered once daily for 91 d; daily feed allotments (overall mean = 9.11 kg dry matter (DM)/d) were generally consumed entirely within 9 h of feeding. Nutrient intakes were not affected by push-up frequency or the interaction of main effects, but all intakes were affected by feedbunk access, except for DM and neutral detergent fiber expressed as a percentage of BW (overall means = 1.93 and 0.80%, respectively). In each case, intakes for FUL were greater than those observed for RES; for DM intake, this amounted to a difference of 0.20 kg/d between those main-effect treatments. After 91 d, heifers without feedbunk restriction exhibited greater final BW, but total gain and average daily gain differed only numerically between FUL and RES. Under the conditions of this trial, heifers were blocked by weight, such that BW were relatively uniform within each pen, and head-locking feed gates were used, which also provided some protection from adjacent aggressive heifers. These results suggest heifers can exhibit acceptable growth performance on high-forage diets in a limit-feeding program that includes moderate feedbunk restriction provided other forms of stress are minimized.

Effects of parasite concentrations on infection dynamics and proliferative kidney disease pathogenesis in brown trout (Salmo trutta).

Proliferative kidney disease (PKD) is an emerging disease of salmonids, which is exacerbating with increasing water temperature. Its causative agent, the myxozoan parasite Tetracapsuloides bryosalmonae, exploits freshwater bryozoans as primary hosts and salmonids as intermediate hosts. Our experiments showed that the manipulation of exposure concentrations of infective malacospores had relatively minor impacts for the disease outcomes in the fish host. In this study, brown trout (Salmo trutta) were exposed to three different exposure concentrations of T.bryosalmonae malacospores: (a) a single low parasite concentration (LC), (b) a single high parasite concentration (HC) and (c) three times a low concentration (repeat exposure, RE). Parasite dynamics in the fish host and release of fish malacospores were quantified and fish kidney histopathology was evaluated to determine PKD pathogenesis. Infection prevalence was always lower in the LC group than in the other groups over the course of the study. While the parasite proliferation phase was slower in the LC group, the maximum parasite burden did not differ significantly amongst treatments. The onset of fish malacospore release (day 45 post-exposure), indicated by detection of T.bryosalmonae DNA in the tank water, occurred at the same time point for all groups. Reduced intensity of kidney pathological development was observed in the LC treatment indicating lower disease severity. While the LC treatment resulted in reduced outcomes across several infection parameters (infection prevalence, parasite proliferation, total fish malacospores released), the overall differences were small. The RE and HC treatment outcomes were for most parameters comparable. Our results suggest that repeated exposure, as is likely to occur in the wild during the summer months, might play a more important role in the dynamics of PKD as an emerging infectious disease than the actual concentration of spores.

Reactions of Rare‐Earth Metal Based Lewis Pairs with Azides†

Summary of main observation and conclusionThe reactivity of homoleptic rare‐earth metal aryloxide based Lewis pairs toward organic azide substrates has been investigated herein. Treatment of RE(OAr)3 (RE = La, Sm, Y, and Sc, Ar = 2,6‐tBu2‐C6H3), PEt3 and Me3SiN3 in 2 : 1 : 1 molar ratio resulted in formation of separated ion pair complexes [Me3Si‐PEt3]+[(ArO)3RE‐N=N=N‐RE(OAr)3]– under mild conditions. Replacement of phosphine with the nitrogen‐containing Lewis base 1,4‐diazabicyclo[2.2.2]octane (DABCO) produced analogous rare‐earth azide complexes with [Me3Si‐DABCO]+ counterions. In contrast, reaction of the La(OAr)3/PEt3 Lewis pair with 1‐adamantyl azide (AdN3) afforded the typical frustrated Lewis pair‐type 1,1‐addition product. A tetrahydrofuran ring‐opening reaction was also observed for the resulting rare‐earth azide complex with the [Me3Si‐PEt3]+ cation, with cleavage of the C—O bond by Si/P cooperation.

Mulching practices alter the bacterial-fungal community and network in favor of soil quality in a semiarid orchard system

The study aimed to investigate how mulching practices can improve soil quality by altering microbial community composition and the interrelations among its members. The experiment was conducted for 7 years (from 2012 to 2018) in an apple (Malus domestica cv. Fuji) orchard located on the Loess Plateau, China. Four treatments were applied: conventional tillage (CT), intercrop ryegrass (Lolium perenne L.) cover (RE), inter-row cornstalk mulch (CS), and black ground fabric mulch (BF). The results indicated that compared to CT treatment, all the three mulching practices significantly increased soil moisture content. The RE and CS treatments improved soil organic matter content by 11.8% and 36.5%, respectively, and CS treatment also increased available nitrogen content in soil by 42.0% compared with CT treatment. The soil environmental heterogeneity under different mulching practices shaped the microbial community structure and the dominant populations. All the mulching practices significantly increased the operational taxonomic unit (OTU) abundance and alpha diversity of fungi, and positively affected bacterial richness. Network analyses showed that the microbial structure was significantly affected by soil water and microbial biomass nitrogen. Interestingly, based on network correlation analysis among microorganisms, the BF treatment reduced competition between bacteria, whereas RE treatment increased their cooperation, however, the CS treatment not only promoted cooperation between fungi, but also increased correlations between fungi and bacteria. Finally, our results indicated that mulching practices not only increased the diversity of microorganisms and abundance of dominant species, but also promoted the interrelations among microorganisms in favor of soil quality.

Abstract No. 562 Temporary reversal of hepatoenteric collaterals during Y90 planning and treatment

A serious complication of Y90 radioembolotherapy (Y90 RE) is nontarget embolization. Current conventional methods to prevent this include coil embolization of hepatoenteric arteries. Temporary redirection of blood towards the liver within these arteries using a balloon catheter in the common hepatic artery provides a safe alternative with prevention of nontarget deposition of Y90 microspheres. Between September 2010 to September 2015, 88 patients with diagnostic or treatment angiograms with attempted use of the balloon occlusion technique were catalogued. A balloon catheter was inflated in the common hepatic artery to temporarily redirect flow from hepatoenteric arteries toward the liver during both diagnostic Tc99m MAA (67 patients) and therapeutic Y90 RE (72 patients) delivery. SPECT/CT nuclear scintigraphy was performed after delivery of each radionuclide. Clinical and laboratory follow-up at 1, 3 and 6 months and imaging follow-up at 3 and 6 months were performed. 72 Y90 RE treatment sessions were performed with use of the balloon technique with either glass (21) or resin (51) microspheres. Tumor types included hepatocellular (26), neuroendocrine (26), colorectal (23), or other (13). Post Y90 RE scintigraphy confirmed no significant extrahepatic activity in any patient. Whole liver, right lobe, left lobe Y90 RE treatments were performed in 58, 15 and 5 sessions respectively. One grade 2 gastrointestinal ulcer was present at 90 day follow-up. Greater than 80% administration of the prescribed dose of was possible in 34/51 resin microsphere sessions and in 20/21 sessions of glass microspheres. Temporary occlusion of the common hepatic artery with a balloon-catheter is a reliable and reproducible alternative to conventional coil embolization of hepatoenteric arteries during both diagnostic Tc99m MAA and therapeutic Y90 RE delivery.

Abstract No. 567 Yttrium-90 radioembolization therapy for combined hepatocellular and cholangiocarcinoma

To report outcomes of transarterial radioembolization (TARE) using glass microspheres for the treatment of mixed hepatocellular-cholangiocarcinoma (HCC-CC). Between 2013 and 2019, 10 consecutive patients with histologically confirmed HCC-CC received 1 or 2 TARE treatments of a targeted territory using glass microspheres. Baseline demographics in addition to tumor distribution, Child Pugh score, and BCLC were recorded. Tumor response was assessed according to modified RECIST criteria. Six-month mortality and adverse events were monitored for each patient. Adverse events were graded by standardized criteria using the Common Terminology Criteria for Adverse Events (CTCAE) v4.0. Eight (80%) patients (4 males, 4 females) received a single TARE treatment and two (20%) patients underwent two. Tumor responses at 6 months were complete and partial in 80% and 20% patients respectively. No patients demonstrated tumor progression. No major toxicity was observed after 1st or 2nd treatment of RE. Two grade 2 adverse events were seen (Table 1). 6-month mortality was 10% (n = 1). While outcomes of combined HCC-CC are poor and optimal treatment remains undefined, TARE appears to represent a safe and promising locoregional treatment for disease control with an acceptable toxicity profile.Table 1Patient #Adverse Events after 1st Procedure (CTCAE Grade)Adverse Events after 2nd Procedure (CTCAE Grade)6 Month Mortality1 Year Mortality2 Year MortalityChild Pugh ScoreBCLCTumor LocationRECIST Rating1NoneNoneYYY6 (A)BRPartial2Abdominal Pain and Fatigue (2)NoneNNN5 (A)BLComplete3Fever (2)NoneNNN5 (A)ARComplete4NoneNoneNNN5 (A)ABiliary DuctComplete5NoneNoneNNN6 (A)ALComplete6NoneNoneNNN5 (A)ARComplete7NoneNoneNNN5 (A)ARPartial8NoneNoneNNN5 (A)ARComplete9NoneNoneNNN5 (A)ARComplete10NoneNoneNNN5 (A)ALComplete Open table in a new tab

Resveratrol protects H9c2 cells against hypoxia-induced apoptosis through miR-30d-5p/SIRT1/NF-κB axis

Previous studies have demonstrated the cardioprotective role of resveratrol (Res). However, the underlying molecular mechanisms involved in the protective role of Res are still largely unknown. H9c2 cells were distributed into five groups: normal condition (Control), DMSO, 20 mMRes (dissolved with DMSO), hypoxia (Hyp), and Res+Hyp. Cell apoptosis was evaluated using flow cytometry and protein analysis of cleaved caspase 3 (cle-caspase 3). qRT-PCR assay was performed to measure the expression of microRNA-30d-5p (miR-30d-5p). MTT assay was performed to evaluate the cell proliferation. The relationship between miR-30d- 5p and silent information regulator 1 (SIRT1) was confirmed by luciferase reporter, RNA immunoprecipitation (RIP), and western blot assays. Western blot was performed to analyze NF-κB/p65 and I-κBα expressions. Our data showed that hypoxia enhanced apoptosis and NF-κB signaling pathway, which was alleviated by Res treatment. Hypoxia increased the expression of miR-30d-5p while decreased the SIRT1expression, which was also attenuated by Res treatment. Furthermore, miR-30d-5p depletion inhibited the proliferation, reduced apoptosis and decreased the expression of cle-caspase 3 in H9c2 cells with hypoxia treatment. Luciferase reporter, RIP, and western blot assays further confirmed that miR-30d-5p negatively regulated the expression of SIRT1. Interestingly, the rescue-of-function experiments further indicated that knockdown of SIRT1 attenuated the effect of miR-30d-5p depletion on proliferation, apoptosis NF-κB signaling pathway inH9c2 cells with hypoxia treatment. In addition, the suppression of NF-κB signaling pathway increased cell viability while decreased cell apoptosis in hypoxia-mediatedH9c2 cells. Our data suggested Res mayprotectH9c2 cells against hypoxia-induced apoptosis through miR-30d-5p/SIRT1/NF-κB axis.

Pak2 inhibition promotes resveratrol-mediated glioblastoma A172 cell apoptosis via modulating the AMPK-YAP signaling pathway.

As a polyphenolic compound, resveratrol (Res) is widely present in a variety of plants. Previous studies have shown that Res can inhibit various tumors. However, its role in c remains largely unexplored. In the present study, we first demonstrated that Res inhibited cell viability and induced apoptosis of glioblastoma A172 cell. Further experiments showed that Res induced mitochondrial dysfunction and activated the activity of caspase-9. Functional studies have found that Res treatment is associated with an increase in the expression of Pak2. Interestingly, inhibition of Pak2 could further augment the proapoptotic effect of Res. Mechanistically, Pak2 inhibition induced reactive oxygen species overproduction, mitochondria-JNK pathway activation, and AMPK-YAP axis suppression. However, overexpression of YAP could abolish the anticancer effects of Res and Pak2 inhibition, suggesting a necessary role played by the AMPK-YAP pathway in regulating cancer-suppressive actions of Res and Pak2 inhibition. Altogether, our results indicated that Res in combination with Pak2 inhibition could further enhance the anticancer property of Res and this effect is mediated via the AMPK-YAP pathway.

Resveratrol corrects aberrant splicing of RYR1 pre-mRNA and Ca2+ signal in myotonic dystrophy type 1 myotubes.

Myotonic dystrophy type 1 (DM1) is a spliceopathy related to the mis-splicing of several genes caused by sequestration of nuclear transcriptional RNA-binding factors from non-coding CUG repeats of DMPK pre-mRNAs. Dysregulation of ryanodine receptor 1 (RYR1), sarcoplasmatic/endoplasmatic Ca2+-ATPase (SERCA) and α1S subunit of voltage-gated Ca2+ channels (Cav1.1) is related to Ca2+ homeostasis and excitation-contraction coupling impairment. Though no pharmacological treatment for DM1 exists, aberrant splicing correction represents one major therapeutic target for this disease. Resveratrol (RES, 3,5,4′-trihydroxy-trans-stilbene) is a promising pharmacological tools for DM1 treatment for its ability to directly bind the DNA and RNA influencing gene expression and alternative splicing. Herein, we analyzed the therapeutic effects of RES in DM1 myotubes in a pilot study including cultured myotubes from two DM1 patients and two healthy controls. Our results indicated that RES treatment corrected the aberrant splicing of RYR1, and this event appeared associated with restoring of depolarization-induced Ca2+ release from RYR1 dependent on the electro-mechanical coupling between RYR1 and Cav1.1. Interestingly, immunoblotting studies showed that RES treatment was associated with a reduction in the levels of CUGBP Elav-like family member 1, while RYR1, Cav1.1 and SERCA1 protein levels were unchanged. Finally, RES treatment did not induce any major changes either in the amount of ribonuclear foci or sequestration of muscleblind-like splicing regulator 1. Overall, the results of this pilot study would support RES as an attractive compound for future clinical trials in DM1. Ethical approval was obtained from the Ethical Committee of IRCCS Fondazione Policlinico Universitario A. Gemelli, Rome, Italy (rs9879/14) on May 20, 2014.

Biological implications of PTEN upregulation and altered sodium/iodide symporter intracellular distribution in resveratrol-suppressed anaplastic thyroid cancer cells.

Objective: Anaplastic thyroid cancer/ATC is a highly aggressive malignancy with extremely poor prognosis. Resveratrol/Res promotes re-differentiation of cancer cells and exerts inhibitory effects on ATC cells. Sodium/iodide symporter/NIS and phosphate and tension homology deleted on chromsome ten/PTEN levels are positively correlated with the grade of thyroid cancer differentiation, while the impact of Res on them remain unknown.Materials and Methods: The patterns of NIS and PTEN expression and intracellular distribution in THJ-16T and THJ-21T ATC and Nthy-ori 3-1 normal thyroid cells and their relevance with Res-caused ATC suppression were investigated via multiple experimental methods. E-cadherin was cited as a re-differentiation biomarker of ATC cells.Results: MTT and EdU cell proliferation assays showed distinct growth suppression in ATC cells after Res treatment. TUNEL staining revealed extensive apoptosis of Res-treated THJ-16T and THJ-21T rather than Nthy-ori 3-1 cells. Western blotting, immunocytochemical/ICC and double-labeled immunofluorescent/IF staining showed increased PTEN levels accompanied with distinct NIS and PTEN nuclear co-translocation in Res-treated THJ-16T and THJ-21T cells. E-cadherin but not NIS appeared on the outer membrane.Conclusion: PTEN upregulation and the concurrent NIS and PTEN nuclear translocation in Res-suppressed ATC cells may indicate the better therapeutic outcome and would be a group of beneficial prognostic factors of ATCs.

Resveratrol inhibits LPS-induced inflammation through suppressing the signaling cascades of TLR4-NF-κB/MAPKs/IRF3.

Resveratrol (Res) is a natural compound that possesses anti-inflammatory properties. However, the protective molecular mechanisms of Res against lipopolysaccharide (LPS)-induced inflammation have not been fully studied. In the present study, RAW264.7 cells were stimulated with LPS in the presence or absence of Res, and the subsequent modifications to the LPS-induced signaling pathways caused by Res treatment were examined. It was identified that Res decreased the mRNA levels of Toll-like receptor 4 (TLR4), myeloid differentiation primary response protein MyD88, TIR domain-containing adapter molecule 2, which suggested that Res may inhibit the activation of the TLR4 signaling pathway. It suppressed the expression levels of total and phosphorylated TLR4, NF-κB inhibitor, p38 mitogen-activated protein kinase (MAPK), c-Jun N-terminal kinase, extracellular signal-regulated kinase 1/2 and interferon (IFN) regulatory factor 3 (IRF3) proteins. Following treatment with Res or specific inhibitors, the production of pro-inflammatory mediators including tumor necrosis factor-α, interleukin (IL)-6, IL-8 and IFN-β were decreased and the expression of anti-inflammatory mediator IL-10 was increased. These results suggested that Res may inhibit the signaling cascades of NF-κB, MAPKs and IRF3, which modulate pro-inflammatory cytokines. In conclusion, Res exhibited a therapeutic effect on LPS-induced inflammation through suppression of the TLR4-NF-κB/MAPKs/IRF3 signaling cascades.

Resveratrol Downregulates STAT3 Expression and Astrocyte Activation in Primary Astrocyte Cultures of Rat.

Astrocytes respond to all forms of central nervous system (CNS) insults by a process referred to as reactive astrogliosis. Inhibition of astrocyte growth and activation is an important strategy for promoting injured CNS repair. STAT3 (signal transducer and activator of transcription 3) is reported to be a critical regulator of astrogliosis, and resveratrol (RES, a dietary polyphenol) is considered to be a natural inhibitor of STAT3 expression and phosphorylation. In this study, we investigated the effects of RES on STAT3 expression and phosphorylation, and then on the proliferation and activation of astrocytes, a critical process in reactive astrogliosis, in rat primary cultured astrocytes and an in vitro scratch-wound model. RES downregulated the expression levels of STAT3, P-STAT3 and GFAP (glial fibrillary acidic protein) in cultured astrocytes. The positive index of Ki67 was apparently reduced in cultured astrocytes after RES treatment. Meanwhile, cultured astrocyte proliferation and activation were attenuated by RES. Moreover, in the established in vitro scratch-wound model the increased expression levels of STAT3, P-STAT3 and GFAP induced by scratching injury were also clearly inhibited by RES. In addition, the inhibitory effect of RES on cell proliferation was similar to that of AG490 (a selective inhibitor of STAT3 phosphorylation) and abrogated by Colivelin (a STAT3 activator) stimuli. Taken together, our data suggest that RES is able to inhibit reactive astrocyte proliferation and activation mainly via deactivating STAT3 pathway. So RES may have a therapeutic benefit for the treatment of the injured CNS.

Resveratrol decreases CD45+ CD206- subtype macrophages in LPS-induced murine acute lung injury by SOCS3 signalling pathway.

Acute lung injury/acute respiratory distress syndrome (ALI/ARDS) are life‐threatening condition in critically ill patients. Resveratrol (Res), a natural polyphenol, has therapeutic effect in animal model with ALI; however, whether Res attenuates ALI through modulation of macrophage phenotypes in the animal model remains unknown. We in this study treated LPS‐induced murine ALI with 30 mg/kg Res and observed significantly reduced severity of ALI in the Res‐treated mice 48 hours after Res treatment. Neutrophil infiltrates were significantly reduced, accompanied with lower infiltration of CD45+Siglec F− phenotype macrophages, but higher population of CD45+Siglec F+ and CD45+CD206+ alternatively activated macrophages (M2 cells) in the Res‐treated mice with ALI. In addition, the expression of IL‐1beta and CXCL15 cytokines was suppressed in the treated mice. However, Res treatment in mice with myeloid cell‐restricted SOCS3 deficiency did not significantly attenuate ALI severity and failed to increase population of both CD45+Siglec F+ and CD45+CD206+ M2 subtype macrophages in the murine ALI. Further studies in wild‐type macrophages revealed that Res treatment effectively reduced the expression of IL‐6 and CXCL15, and increased the expression of arginase‐1, SIRT1 and SOCS3. However, macrophages’ lack of SOCS3 expression were resistant to the Res‐induced suppression of IL‐6 and CXCL15 in vitro. Thus, we conclude that Res suppressed CD45+Siglec F− and CD45+CD206− M1 subtype macrophages through SOCS3 signalling in the LPS‐induced murine ALI.

Resveratrol inhibits DHT-induced progression of prostate cancer cell line through interfering with the AR and CXCR4 pathway

Prostate cancer (PCa) is one of the most common malignancies and the second most common cause of cancer-related deaths in men world-wide and is known to be affected by the action of dihydrotestosterone (DHT) via androgen receptor (AR). Resveratrol (Res) as a phytochemical in grapes and red wine has diverse biological effects such as anti-inflammation, anti-oxidation and anti-cancer. CXCR4 as a chemokine receptor has been found to be upregulated in cancer metastasis and has been used as a prognostic marker in various types of cancer, including leukemia, breast cancer, and prostate cancer. In this study, we focused on the role of DHT in the induction of prostate cancer progression by affecting the AR and CXCR4 pathway. Also, we investigated the inhibition effect of resveratrol on DHT-induced prostate cancer metastasis. In cell viability assay, DHT increased the cell viability of LNCaP prostate cancer cells, on the other hand, Res and its combination with bicalutamide (BCT) as an AR-antagonist or AMD3100 as a CXCR4 inhibitor significantly reduced the cell viability promoted by DHT. Trans-well migration assay and wound healing assay represented the similar results with cell viability assay. According to the results of TUNEL assay, the apoptotic activity was induced by treatment of Res. As results of western blot analysis, the expression of AR, CXCR4, p-PI3K, and p-AKT and the downstream genes related with cell cycle progression and epithelial-mesenchymal transition (EMT) were decreased and the expression of the apoptosis-related genes was increased by treatment of Res and its combination with BCT or AMD3100. This study would suggest that Res and its combination with AR and CXCR4 antagonists can be used in order to suppress the metastatic behaviors of prostate cancer.

Anti-obesity Effects of Ginsenosides in High-Fat Diet-Fed Rats.

To examine the anti-obesity effects of ginsenosides in Korea Red Ginseng (KRG, Panax ginseng) in rats fed with a high-fat diet (HFD). Twenty-five 4-week-old obesity rats after receiving an HFD for 5 weeks; subsequently, they were additionally treated with ginsenosides Rb1, Rd, Rg1, or Re (10 mg/kg, intraperitoneal injection) for a further 3 weeks (n=5 in each group). The control rats were fed a normal diet. The food consumption, body weight, locomotor activity, serum lipids, adipose tissues, nitric oxide (NO) expression, leptin, neuropeptide Y (NPY), cholecystokinin (CCK) in the brains were measured. In the HFD-fed rats, body weight, body fat mass, serum levels of leptin and NO were significantly higher than in the control rats (P<0.05 or P<0.01). However, the treatment of Rd, Re, and Rb1 markedly decreased body fat mass and body weight (P<0.05). The serum level of leptin and NO in ginsenoside-treated rats were markedly lower than the control group (P<0.01). The expression of NPY and CCK in the hypothalamic nuclei showed insignificant difference among groups. However, the expression of NPY immunoreactive neurons in the hypothalamus was significantly reduced in the Rb1-treated group (P<0.05). PD-type ginsenoside Rb1 from the crude saponins of KRG may be a useful compound for the treatment of obesity and related disorders through the modulation of peripheral and central appetite-regulating signals.

Resveratrol improved the developmental potential of oocytes after vitrification by modifying the epigenetics.

Resveratrol (Res) has been reported to be able to improve oocyte vitrification because of its antioxidative properties. The objective of this study was to further assess the positive effect of Res addition on the developmental potential of vitrified mouse oocytes from the perspective of epigenetic alterations. First, 2 μM Res was chosen as the optimal concentration on the basis of its effects on survival and its antioxidative properties. We found that Res addition significantly promoted fertilization (63.8% vs. 42.9%) and blastocyst formation (68.3% vs. 50.2%) after oocyte vitrification. The quality of the derived blastocysts was also higher after Res treatment. Regarding epigenetic aspects, the expression of the important deacetylase SIRT1 was found to decrease significantly upon vitrification, but it was rescued by Res. The abnormal levels of H3K9 acetylation and DNA methylation in vitrified oocytes were restored by Res addition. Moreover, the expression of several imprinted genes was affected by oocyte vitrification. Among them, abnormal Gtl2 and Peg3 expression levels were restored by Res addition. Therefore, the methylation of theirimprinted control regions (ICRs) was examined. Surprisingly, the abnormal patterns of Gtl2 and Peg3 methylation in blastocysts developed from vitrified oocytes were both restored by Res addition. Finally, the full-term embryonic development showed that the birth rate was improved significantly by Res addition (56.2% vs. 38.1%). Collectively, Res was beneficial for the pre- and postimplantation embryonic development. Except for the antioxidative activity, Res also played a role in the correction of some abnormal epigenetic modifications caused by oocyte vitrification.

Resveratrol Ameliorates Glucocorticoid-Induced Bone Damage in a Zebrafish Model.

Resveratrol (Res) is a multi-functional polyphenol compound that has protective functions in cardiovascular and neurodegenerative diseases. This study aimed to determine the effect of Res on osteogenic differentiation and bone mineralization in zebrafish (Danio rerio) with dexamethasone (Dex)-induced bone damage. Our results showed that Dex exposure (15 μmol/l) decreased the green fluorescence areas and the integrated optic density (IOD) values in the skull bones of zebrafish larvae of the TG(SP7:EGFP) strain in a dose-dependent manner (p < 0.01). Furthermore, Dex exposure decreased the alizarin red S-stained areas (bone mineralization area) in the skeleton and spinal bones of zebrafish larvae of the AB strain in a dose-dependent manner (p < 0.01). By contrast, Res treatment (150 μmol/l) significantly increased both the green fluorescence and bone mineralization area in Dex-exposed zebrafish larvae. Thus, our data show that Res improves bone mineralization after glucocorticoid-induced bone damage in a zebrafish model. Res may be a candidate drug for the prevention of osteoporosis.

Potentiation of paclitaxel effect by resveratrol in human breast cancer cells by counteracting the 17β-estradiol/estrogen receptor α/neuroglobin pathway.

Neuroglobin (NGB), an antiapoptotic protein upregulated by 17β-estradiol (E2), is part of E2/estrogen receptor α (ERα) pathway pointed to preserve cancer cell survival in presence of microenvironmental stressors including chemotherapeutic drugs. Here, the possibility that resveratrol (Res), an anticancer plant polyphenol, could increase the susceptibility of breast cancer cells to paclitaxel (Pacl) by affecting E2/ERα/NGB pathway has been evaluated. In MCF-7 and T47D (ERα-positive), but not in MDA-MB 231 (ERα-negative) nor in SK-N-BE (ERα and ERβ positive), Res decreases NGB levels interfering with E2/ERα-induced NGB upregulation and with E2-induced ERα and protein kinase B phosphorylation. Although Res treatment does not reduce cell viability by itself, this compound potentiates Pacl proapoptotic effects. Notably, the increase of NGB levels by NGB expression vector transfection prevents Pacl or Res/Pacl effects. Taken together, these findings indicate a new Res-based mechanism that acts on tumor cells impairing the E2/ERα/NGB signaling pathways and increasing cancer cell susceptibility to chemotherapeutic agent.