The post-thaw quality, antioxidant activity, and in vivo fertility of Zaraibi buck semen frozen-stored in the presence of different concentrations of either quercetin or L-arginine.

BackgroundTendon adhesion is one of the most common clinical problems, which poses a considerable challenge to orthopedics doctors. Quercetin (QUE) as a popular drug at present, it has various biological functions, including anti-inflammatory, anti-ischemic, anti-peroxidation, and antioxidant. The purpose of this study was to investigate the effect of quercetin on tendon adhesion and whether quercetin can inhibit oxidative stress.MethodThirty-six rats were randomly divided into three groups, including control group, low QUE (50 mg/kg/day) group, and high QUE (100 mg/kg/day) group. After 1 week, the levels of SOD, MDA and GPx were measured. The degree of tendon adhesion was assessed by macroscopic evaluation and histological evaluation. After 4 weeks. Besides, the pharmacological toxicity of quercetin to main organs were evaluated by histological analysis.ResultsThe extent of superoxide dismutase (SOD) and glutathione peroxidase (GPx) of tendon tissue in high QUE group was significantly higher than those of low QUE group and control group. And the extent of malondialdehyde (MDA) of tendon tissue in high QUE group was significantly lower than that of low QUE group and control group. By macroscopic evaluation and histological analysis, the extent of tendon adhesion in high QUE group was lower than low QUE group and control group. However, there were no significant changes of the major organs through histological analysis.ConclusionsQuercetin may be a good and safe strategy in preventing tendon adhesion. But further clinical research is needed before its recommendation in the prevention and treatment of tendon adhesion.

Objective The study was to investigate the effects of different concentrations of quercetin on bone marrow mesenchymal stem cells (BMSCs) -induced immune tolerance in rats after kidney transplantation. Methods he rats were randomly divided into four groups after kidney transplantation: control group, BMSCs group, MSC+ low-dosage quercetin group and MSC+ high-dosage quercetin group. Janus-activated kinase (JAK) , Signal transducer and activator of transcription 3 (STAT3) , p-STAT3 protein expression in kidney tissue were detected by Western blot. Serum transforming growth factor-β1 (TGF-β1) was detected by ELISA assay. We collected the proximal tubule epithelial cells from the MSC group, and cultured them with or without stimulation of quercetin. Expressions of JAK, STAT3, p-STAT3, and TGF-β1 were detected by Western blot after STAT3 was inhibited and overexpressed in these cells, respectively. Results After kidney transplantation, TGF-β1 in serum in the controlgroup (19 162±223) pg/ml was higher than any of other groups (BMSC group, 12 106±163 pg/ml; MSC+ high dosage of quercetin groups, 9 113±110 pg/ml; MSC+ low dosage of quercetin groups, 9 024±133 pg/ml) . When compared to the control group, theqvalues of these groups were 96.88, 138.0, 139.2, respectively (P < 0.01) . Protein expression of JAK and p-STAT3 in the kidney tissue (1.5±0.2, 10.7±0.5) were significantly higher in control group than in the other groups: BMSC group (0.9±0.3, 8.5±0.2;P = 0.0059) , MSC+ high dosage of quercetin groups (0.7±0.2, 6.1±0.3;P = 0.0002) , and MSC+ low dosage of quercetin groups (0.7±0.3, 5.8±0.1;P = 0.0011) . In group with inhibition of STAT3, relative protein expression of p-STAT3 and TGF-β1 (0.7±0.1, 0.3±0.1) in the proximal tubule epithelial cells were significantly reduced compared to the other groups: control group (8.2±0.4, 7.1±0.5;P < 0.0001,P < 0.0001) , quercetin group (5.6±0.3, 4.8±0.4;P < 0.0001,P < 0.0001) , and si-control group (5.7±0.3, 4.5±0.4;P < 0.0001,P < 0.0001) . In group with overexpression of STAT3, relative protein expression of p-STAT3and TGF-β1 (10.5±0.6, 8.4±0.5) were significantly higher than those in the quercetin group (5.7±0.3, 5.4±0.3;P < 0.0001,P < 0.0001) and vector plasmid group (6.1±0.1, 5.6±0.4;P < 0.0001,P = 0.0001) . Conclusion Simultaneous administration of quercetin and transplantation of bone marrow mesenchymal stem cells can effectively inhibit rejection and improve renal function in rats with kidney transplantation. Quercetin may regulate immune function through JAK/STAT3/ TGF-β1 pathway. Key words: Quercetin; Bone marrow; Mesenchymal stem cells; Kidney transplantation; Immune tolerance

ObjectiveThis study was designed to investigate the hypothesis that dietary quercetin (QUE) and coated sodium butyrate (SB) supplementation alleviate oxidative stress in the small intestine of diquat (DIQ)-challenged pullets.MethodsA total of 200 13-week-old pullets were divided into four groups: the control group (CON), the DIQ group, the QUE group, and the coated SB group, and injected intraperitoneally with either saline (CON) or diquat (DIQ, QUE, and SB) to induce oxidative stress on day 0.ResultsOn the first day, the malondialdehyde and superoxide dismutase (SOD) concentrations in the SB group were significantly different from those in the DIQ and QUE groups (p<0.05), and dietary supplementation with SB increased serum glutathione peroxidase (GSH-PX) levels compared with the DIQ group (p<0.05). Quercetin and SB increased the levels of CLAUDIN-1 and zonula occludens-1 (ZO-1) in the jejunum. On the tenth day of treatment, QUE attenuated the decrease in GSH-PX levels compared to those of the CON group (p<0.05), while SB increased SOD, GSH-PX, and total antioxidant capacity levels compared to those of the DIQ group. Nuclear factor erythroid 2-related factor 2 (NRF2) and heme oxygenase-1 (HO-1) mRNA levels in the QUE and SB groups increased (p<0.05) and CLAUDIN-1 mRNA levels in the QUE and SB groups were upregulated compared to those in the DIQ group ileum tissue.ConclusionSupplementation of QUE and SB demonstrated the ability to relieve oxidative stress in pullets post DIQ-injection with a time-dependent manner and QUE and SB may be potential antioxidant additives for relieving oxidative stress and protecting the intestinal barrier of pullets.

Background Oxidative stress-induced apoptosis of human lens epithelial cells (LECs) is associated with c-Jun N terminal kinase (JNK) pathway.Quercetin possesses the antioxidation by inhibiting the JNK pathway.However,whether quercetin can protect LECs from the oxygen-induced damage is still not proved.Objective This study attempted to invatigate the effects and its mechanism of quercetin against hyperbaric oxygeninduced LECs apoptosis. Methods Human LECs line SRA01/04 was cultivated and passaged in MEM medium containing 10％ fetal bovine serum and 0.5％ non-essential amino acids for 2 hours,with or without 20 μmol/LSP600125 or 1 μmol/L quercetin prior to exposure to hyperbaric oxygen.Each exposure session remained 6 hours in 99％ O2 and 1％CO2 with a pressure chamber at 588 kPa.The viability of human LECs was detected by MTT.Cell apoptosis was assessed by flow cytometer using Annexin V-FITC apoptosis detection.The expression of JNK/p-JNK,c-Jun/p-c-Jun,caspase 3 and caspase 9 were detected by Western blot. Results LECs viability (A570 ) was 0.835 ±0.082,0.450±0.083,0.654±0.079,0.649±0.090 respectively in the blank control group,hyperbaric oxygen exposed group,hyperbaric oxygen+SP600125 group and hyperbaric oxygen+quercetin group.The A570 in the hyperbaric oxygen exposed group was significantly lower than the blank control group ( P =0.000),but those in hyperbaric oxygen + SP600125 group and hyperbaric oxygen+quercetin group were significantly higher than the hyperbaric oxygen exposed group ( P =0.003,0.002 ).The numbers of apoptosis cells were 3.17 ±0.74,19.77 ± 1.44,8.45 ±0.93,7.79 ±0.78 respectively in the blank control group,hyperbaric oxygen exposed group,hyperbaric oxygen+SP600125 group and hyperbaric oxygen+quercetin group.Apoptotic LECs were significantly increased in the hyperbaric oxygen exposed group compared with the blank control group ( P=0.000),but those in the hyperbaric oxygen+SP600125 group and hyperbaric oxygen+quercetin group were significantly reduced in comparison with hyperbaric oxygen exposed group (both P=0.000).In additional,expressions of p-JNK,p-c-Jun,caspase 3 and caspase 9 proteins in the cells were elevated in the hyperbaric oxygen exposed group compared with the blank control group (all P =0.000 ),however,those in the hyperbaric oxygen + SP600125 group and hyperbaric oxygen + quercetin group were declined when compared with the hyperbaric oxygen exposed group( all P＜0.05 ). Conclusions JNK pathway is involved in the apoptotic procedure of human LECs induced by oxygen stress.SP600125 and certain concentration of quercetin can interdict the JNK signal pathway and endogenous apoptosis of LECs and further alleviate hyperbaric oxygen-induced damage of LECs. Key words: Quercetin; Hyperbaric oxygen; Lens epithelial cell; c-Jun N-terminal kinases; Apoptosis

To detect the expression of heparanase (HPA) in cervical cancer cells and investigate the impact of quercetin on the expression of HPA, and the molecular mechanism that quercetin inhibits the growth of cervical cancer cells. The experimental groups included cervical cancer cell lines (HeLa and Caski) exposed to different concentrations of quercetin (20, 40 and 80 µmol/L) in the culture medium. The control groups included a negative control group, which was cultured with RPMI 1640 only, and a positive control group, in which cervical cancer cells were transfected with HPA small interference RNA (siRNA) to silence HPA expression. The cellular expression levels of HPA were detected with fluorescence quantitative real-time PCR and western blot analysis at 24, 48 and 72 hours after treatment. (1) HPA was significantly expressed in both cervical cancer cell lines (HeLa and Caski), and it exists both nucleus and cytoplasm. (2) The real-time PCR shows as follows: as the quercetin concentration increased (20, 40 and 80 µmol/L), the mRNA expression level of HPA decreased (P < 0.01), in which the inhibition of HPA expression was concentration dependent. In addition, the inhibition of HPA expression was also time dependent. As time growth, the expression level of HPA mRNA (24, 48 and 72 hours) in HeLa and Caski cells decreased (P < 0.01). Compared with negative control group, the expression level of HPA mRNA decreased in different concentrations of quercetin (40 and 80 µmol/L) in both HeLa and Caski cells (all P < 0.05); Compared with positive control group, the expression level of HPA mRNA expressed no obvious difference in quercetin (80 µmol/L) group (P > 0.05) in HeLa cells, while it was opposite in Caski cells (P < 0.01). (3) The result of western blot shown that, as the quercetin concentration increased (20, 40 and 80 µmol/L) and time growth (24, 48 and 72 hours), the expression level of HPA protein decreased (P < 0.01), and the inhibition of HPA protein expression was concentration and time dependent. Compared with negative control group, the expression level of HPA protein decreased in different concentrations of quercetin (40 and 80 µmol/L) in both HeLa and Caski cells (all P < 0.05); Compared with positive control group, the expression level of HPA protein expressed no obvious difference in quercetin (80 µmol/L) group (all P > 0.05) in both HeLa cells and Caski cells (all P > 0.05). Quercetin could inhibits the expression of HPA in cervical carcinoma cell lines, which inhibition is concentration and time dependent.

The present study aimed to examine the protective effect of quercetin (QUE) on cyclophosphamide (CTX)-induced nephrotoxicity. For that purpose, 24 mice were divided into four groups (Control, QUE, CTX, and CTX + QUE). The CTX and CTX + QUE groups received 200 mg/kg of cyclophosphamide on the 1st and 7th days. The QUE and CTX + QUE groups were treated with 50 mg/kg of quercetin daily for 14 days. At the end of the experiment, the animals were sacrificed, and kidney samples were analyzed. The results indicated that CTX leads to severe morphological degenerations and disruption in renal function. Serum BUN, Creatinine, Uric acid, tissue Bax, Caspase 3, TNF-α and IL-1β expression levels were upregulated in the CTX group compared to Control and QUE groups (p < 0.05). Although MAPK/ERK phosphorylation level is not affected in CTX group, there was a significant increase in CTX + QUE group (p < 0.05), but the NF-κB was significantly suppressed in this group (p < 0.01). The RT-qPCR results showed that the cyt-c and the Bax/Bcl-2 ratio mRNA expression folds were upregulated in the CTX group (p < 0.01), which was downregulated in the CTX + QUE group. However, there was a significant difference in the CTX + QUE group compared to the Control and QUE groups (p < 0.01). The findings showed that administering quercetin along with cyclophosphamide alleviated renal injury by regulating apoptotic and inflammatory expression. Moreover, the administration of quercetin and cyclophosphamide could synergistically improve renal function test results, and activate cellular responses, which upmodulate MAPK/ERK phosphorylation and suppression of NF-κB.

Background: Taurine supplementation in tris egg yolk citrate (TEYC) extender can improve antioxidant defense and reduce motility degeneration rate in semen of Surti buck preserved at refrigeration temperature. Present study has evaluated antioxidant effect of different concentrations of taurine in TEYC extender on oxidative stress, antioxidant defense and motility degeneration rate in Surti buck semen preserved at refrigerated temperature. Methods: Four Surti bucks (age greater than 2years) were selected. Total 64 ejaculates (16 per buck) were collected during a period of 8 week. Post-collection they were pooled and stored at refrigeration temperature after dividing into 5 groups based on different taurine concentrations in TEYC extender viz. 0 mM (control T1), 25 mM (T2), 50 mM (T3), 75 mM (T4) and 100 mM (T5) taurine maintaining final concentration of 200 x 106 sperm/ml (pH 6.5-6.8). Evaluation of motility degeneration rate at 24, 36 and 48 hours and glutathione as well as lipid peroxidation (MDA) at 0 and 48 hours was done. Result: Supplementation of taurine @50 mM in TEYC extender caused post-chilling significant increase in reduced glutathione (GSH) levels, lowering of lipid peroxidation (in terms of MDA production) and reduction of motility degeneration rate (MDR)% in semen of Surti bucks. It improved antioxidant defense thereby maintaining good quality of semen.

To investigate the effect of lead exposure on copper and copper metalloenzyme and the intervention effect of quercetin. Twenty-four specific pathogen-free male Sprague-Dawley rats of good health were randomly divided into control group (n = 8), lead acetate group (n = 8), and lead acetate + quercetin group (n = 8). The rats in lead acetate group were poisoned by drinking water with 1 g/L lead acetate for 8 weeks, while the rats in control group were fed by drinking water with sodium acetate of the same volume for 8 weeks; the rats in lead acetate+quercetin group were intraperitoneally injected with quercetin (30 mg × kg-1 × d-1) for 8 weeks while drinking water with lead acetate. The Morris water maze was used to test the learning and memory abilities of rats. The lead and copper levels in the serum, hippocampus, cortex, and bone were measured by graphite furnace atomic absorption spectrometry. The level of advanced glycation end products, activity of Cu/Zn superoxide dismutase (SOD), and content and activity of ceruloplasmin (CP) in the hippocampus and serum were measured using a test kit. HE staining was performed to observe the pathological changes in the hippocampus. The Morris water maze test showed that the latency in lead acetate group (52.50±12.04 s) was significantly longer than that in control group (28.08±7.31 s) (P<0.05), and the number of platform crossings was significantly lower in the lead acetate group than in the control group. Compared with those in the control group, the lead levels in the cortex and hippocampus in lead acetate group increased 2.72-fold and 3.79-fold, and the copper in the cortex and hippocampus, and serum free copper levels in lead acetate group increased 1.15-fold, 1.48-fold, and 6.44-fold. Compared with the control group, the lead acetate group had a lower content of CP in the hippocampus (1.23±0.40 U/mg provs0.78±0.08 U/mg pro) and 31.81%and 19.49%decreases in CP content and Cu/Zn SOD activity. Free copper level in serum was positively correlated with the latency and lead levels in the serum, cortex, and hippocampus. The escape latency of rats in lead acetate + quercetin group was decreased by 42.15% (P<0.05). The lead levels in the cortex and hippocampus in lead acetate + quercetin group (0.246 ± 0.58 µg/g and 0.202±0.049 µg/g) were significantly lower than those in lead acetate group (0.391±0.49 µg/g and 0.546±0.120 µg/g), but the free copper and copper levels in the hippocampus and cortex were not significantly reduced. The lead acetate + quercetin group had higher Cu/Zn SOD activity and CP content in the hippocampus than the lead acetate group (P < 0.05). The light microscope observation showed that the number of cells in the hippocampus was reduced with disordered arrangement in the lead acetate group; with quercetin intervention, the hippocampus damage was reduced. Lead exposure results in disorder of copper homeostasis, while quercetin may alleviate the damage induced by lead to some extent.

Chemotherapy drugs may lead to hepatic injury, which is considered one of the limitations of these drugs. The aim of this study was to evaluate the effect of quercetin (QUE) on M1/M2 macrophage polarization and hepatoprotective effect in cyclophosphamide (CTX)-induced liver toxicity. Twenty-four mice were divided into four groups (Control, QUE, CTX, CTX + QUE). The CTX and CTX + QUE groups received 200mg/kg CTX. The animals in the QUE and CTX + QUE groups received 50mg/kg QUE. All animals were sacrificed, and serum and liver samples were used for laboratory analyses. Examinations indicated that CTX exposure led to disruption of liver functions and morphological degenerations. Tissue pro-apoptotic Bax and caspase 3, pro-inflammatory TNF-α and IL-1β, transcription factor NF-κB, and M1 macrophage polarization marker CD86 were upregulated significant (p<0.05) in this group. In addition, CTX exposure led to significantly (p<0.05) upregulation of the Bax/Bcl-2 mRNA ratio and DNA fragmentations. The PCNA-positive hepatic cell ratio and anti-apoptotic Bcl-2 expression are remarkably suppressed (p<0.05). Immunohistochemical analyses are also indicated that M2 macrophage polarization marker CD163 is slightly but remarkably (p<0.05) downregulated in the CTX group compared to the Control and QUE groups. The morphological and biochemical disruptions were alleviated in QUE-treated animals in the CTX + QUE group. Liver function test results, apoptosis, inflammatory, transcription factor NF-κB, regeneration/proliferation, and apoptotic index results in this group were similar (p>0.05) to the control and QUE groups. The M1 cell surface marker expression of CD86 is significantly (p<0.05) downregulated, and M2 macrophage polarization marker expression of CD163 is upregulated significantly (p<0.05) compared to the CTX group. This study indicates that QUE has the potential to downregulate CTX-induced hepatic injury and regulate M1/M2 macrophage polarization to the M2 side, which indirectly demonstrates activation of anti-inflammatory signalling and tissue repair.

Quercetin attenuated oxidative DNA damage through NRF2 signaling pathway in rats with DMH induced colon carcinogenesis

OPTIXcell improves the postthaw quality and fertility of buffalo bull sperm

Objective To explore the mechanism of quercetin against the toxicity of amyloid β (Aβ) to neuroblastoma cell SK-N-SH. Methods In the experiment, SK-N-SH cells were divided to four groups, control group, Aβ group, Aβ+ quercetin group and quercetin group, and treated for 72h.The proliferation of cells, the concentrations of reactive oxygen species (ROS) and glutathione (GSH) in cells were determined. Results Compared with the control group, Aβ[Aβ grouP=(50.6±2.5)]could significantly suppress the proliferation of neuroblastoma cells (P=0.000). However, Aβ group added with quercetin[Aβ+ quercetin=(88.0±9.1)]showed decreased cellular toxicity induced by Aβ(P=0.001). Additionally, there was no significant difference between the control group and quercetin group.Likely, the concentration of ROS increased in Aβ group (523.3±22.5), which was higher than (190.3±25.7) in the control group and (224.0±56.0) in quercetin group (P=0.000). At last, quercetin could increase GSH level in SK-N-SH cells and higher than Aβ group[(3.0±0.2), P=0.004]and Aβ+ quercetin[(5.6±1.1), P=0.008]. Conclusion Quercetin could attenuate the suppression of proliferation of SK-N-SH cells induced by Aβ, that might be though increased concentration of GSH and decreased level of ROS.These data suggested that quercetin has clinical potentials in neurology. Key words: GSH; Quercetin; Alzheimerś disease; Amyloidβ

Semen was collected from four Surti bucks (>24 months-old) twice a week for 8 weeks (16 ejaculates per buck) during November, 2020 to March, 2021. Ejaculates of all four bucks were pooled and divided into five equal aliquots. The aliquots were diluted with tris-egg yolk-citrate (TEYC) extender containing 0 (control T1), 25 (T2), 50 (T3), 75 (T4) and 100 mM (T5) taurine with final concentration of 200x106 sperm/ml (pH 6.5 to 6.8) in each. At 24, 36 and 48 h of refrigerated storage, individual sperm motility, per cent live sperm and HOST reacted sperm were significantly (p<0.01) higher in T3 (50 mM) and abnormal sperm % was significantly (p<0.01) lower in T3 (50 mM) and T4 (75 mM) as compared to other groups. With increasing duration of storage, significant (p<0.01) increasing trend for abnormal sperm per cent and decreasing trend for rest of the characters were observed. Motility showed significant (p<0.01) positive correlation with live spermatozoa, abnormal spermatozoa and HOST-reacted spermatozoa. Live count showed significant (p<0.01) negative correlation with abnormal spermatozoa and significant (p<0.01) positive correlation with HOST-reacted spermatozoa. Live spermatozoa showed significant (p<0.01) negative correlation with abnormal spermatozoa and HOST non-reacted spermatozoa. Abnormal spermatozoa showed significant (p<0.01) negative correlation with HOST-reacted spermatozoa. It was concluded that supplementation of 50 mM taurine in TEYC extender maintained optimum quality of buck semen at refrigerated temperature up to 48 h.

The durability of the hybrid layer, which forms the basis of successful bonding in restorative dentistry, depends on the stability of collagen fibrils. Dentin matrix metalloproteinases degrade the collagen structure over time, leading to a reduction in bond strength. Therefore, MMP inhibitors play a crucial role in maintaining bond durability. The study aims to evaluate and compare the short-term (24 h) and long-term (12 months) effects of dentin pretreatment agents -chlorhexidine (CHX), quercetin (QC), naringin (NR), riboflavin (RF), and α-tocopherol (α-TF)- on bond strength. A total of 144 extracted human molars were collected and randomly divided into six groups: control (no treatment), 2% chlorhexidine, 1% QC, 1% Nar, 3% riboflavin, and 10% α-TF. The solutions were prepared and applied to the dentin surfaces for 60 s, followed by the application of All-Bond Universal adhesive and composite resin. Following the bonding procedure, a nanohybrid flowable composite was applied onto the dentin surface of each tooth using a starch tube, forming a cylindrical specimen with a bonding area of 1 mm 2 (1 mm in diameter and 1 mm in height). Half of the specimens in each group (n = 12) were tested for micro shear bond strength (μSBS) after 24 h of storage, while the remaining specimens underwent thermocycling to simulate 12 months of aging before μSBS testing. To determine the μSBS, each bonded specimen was subjected to a μSBS test in a universal testing machine (MOD Dental MIC-101, Esetron Smart Robotechnologies, Ankara, Turkey) equipped with a 5-kN load cell and featuring a lower fixed and an upper movable compartment, operating at a crosshead speed of 0.5 mm/min. After μSBS testing, failure modes were determined using a stereomicroscope. Data were analyzed using one-way analysis of variance (ANOVA) followed by Tukey’s post hoc tests with p ˂ 0.05 as the significance level. Results of the one-way ANOVA test revealed that, at the end of 24 h, the highest μSBS was observed in the QC group, followed by CHX and NG, all of which showed significantly higher bond strength than the α-TF group (p &amp;lt; 0.05). After 12 months of aging, an increase in SBS was observed in the QC, RF, and naringin groups, whereas a decrease was noted in the Con, CHX, and α-TF groups. QC and NG are natural and effective agents that can be alternatives to CHX in dentin pretreatment. QC showed promise in terms of long-term success, especially after thermal aging, showing the highest bond strength.

Synergistic effect of extracellular adenosine triphosphate and quercetin on post-thaw quality and fertilization potential of Lohi ram sperm