Role of salidroside in a mouse model of non-alcoholic fatty liver disease： A study based on nicotinamide phosphoribosyltransferase / 临床肝胆病杂志

ObjectiveTo investigate the protective effect of salidroside against nonalcoholic fatty liver disease （NAFLD） and its mechanism of action. MethodsA total of 24 male KM mice were randomly divided into normal group， HFD group， HFD+blank control group， and HFD+salidroside group， with 6 mice in each group. The mice in the normal group were given normal diet， and those in the other groups were given high-fat diet. After 14 weeks of modeling， the mice were given salidroside 100 mg/kg/day by gavage， and related samples were collected at the end of week 22. Enzyme-linked immunosorbent assay was used to measure the serum levels of related biochemical parameters including alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， triglyceride （TG）， total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， and low-density lipoprotein cholesterol （LDL-C）； HE staining and NAFLD activity score （NAS） were used to observe the liver histopathology of mice； Western blot was used to measure the changes in the expression of NAMPT， Sirt1， AMPKα， and SREBP1 in liver tissue. A one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the normal group， the HFD group had obvious steatosis and extensive large lipid droplets in liver tissue， with significant increases in NAS score （P<0.01） and the content of AST， ALT， TG， TC， and LDL-C in peripheral blood （all P<0.05） and a significant reduction in the content of HDL-C （P<0.05）， as well as significant reductions in the expression levels of NAMPT， AMPKα， and Sirt1 in liver tissue （all P<0.05） and a significant increase in the expression level of SERBP1 （P<0.01）. Compared with the HFD group and the HFD+blank control group， the HFD+salidroside group had reductions in the distribution of vacuolar lipid droplets and intralobular inflammation in liver tissue， alleviation of the ballooning degeneration of hepatocytes， significant reductions in NAS score （P<0.01） and the content of AST， ALT， TG， and LDL-C in peripheral blood （all P<0.05）， and a significant increase in the content of HDL-C （P<0.05）， as well as significant increases in the expression levels of NAMPT， AMPKα， and Sirt1 in liver tissue （all P<0.05） and a significant reduction in the expression level of SERBP1 （P<0.01）. ConclusionSalidroside can significantly improve the pathological state of mice with NAFLD induced by high-fat diet and exert a protective effect against NAFLD by increasing the expression of NAMPT， Sirt1， and AMPKα and reducing the expression of SERBP1.

Role of nicotinamide phosphoribosyltransferase in delaying smooth muscle cell senescence and protecting abdominal aortic aneurysm / 中华危重病急救医学

Objective:To investigate the protective effect of nicotinamide phosphoribosyltransferase (NAMPT) on abdominal aortic aneurysm by delaying the senescence of aortic vascular smooth muscle cells (VSMC).Methods:The primary VSMC cells from normal and patients with abdominal aortic aneurysm were cultured by tissue adherence method. Cells were divided into normal human-derived VSMC group (Ctrl-VSMC group), abdominal aortic aneurysm patient-derived VSMC group (AAA-VSMC group), and angiotensinⅡ(AngⅡ) in vitro abdominal aortic aneurysm model group (AngⅡ-VSMC group, 100 nmol/L AngⅡ treated normal human-derived VSMC for 48 hours), AngⅡ+P7C3 group and AAA+P7C3 group after NAMPT agonist P7C3 intervention (adding 5 μmol/L P7C3 on the basis of AngⅡ-VSMC group and AAA-VSMC group, respectively). Immunofluorescence staining was used to identify VSMC; cell proliferation-associated antigen Ki67 staining was used to detect cell proliferation; senescence associated β-galactosidase (SA-β-gal) staining was used to detect cell senescence in each group; Western blotting was used to detect the protein expression levels of senescence-related proteins p21, p16 and NAMPT in each group. Results:Compared with the Ctrl-VSMC group, the positive rate of SA-β-gal staining and the expression levels of senescence-related proteins p21 and p16 in the AAA-VSMC group and AngⅡ-VSMC group were significantly increased [SA-β-gal staining positive rate: (74.1±4.4)%, (68.6±5.5)% vs. (36.8±10.3)%, p21/GAPDH: 0.61±0.07, 0.51±0.03 vs. 0.31±0.03, p16/GAPDH: 0.77±0.03, 0.72±0.06 vs. 0.33±0.26, all P < 0.01]. However, the expression of NAMPT was significantly decreased (NAMPT/GAPDH: 0.88±0.07, 0.79±0.14 vs. 1.29±0.02, both P < 0.01). Compared with the AngⅡ-VSMC group, the positive rate of SA-β-gal staining and the expressions levels of senescence-related proteins p21 and p16 in the AngⅡ+P7C3 group were significantly lower [SA-β-gal staining positive rate: (49.1±3.2)% vs. (68.6±5.5)%, p21/GAPDH: 0.35±0.06 vs. 0.51±0.03, p16/GAPDH: 0.47±0.08 vs. 0.72±0.06, all P < 0.05], while the expression of NAMPT was significantly increased (NAMPT/GAPDH: 1.15±0.06 vs. 0.79±0.14, P < 0.01). Compared with the AAA-VSMC group, the positive rate of SA-β-gal staining and the expression levels of senescence-related proteins p21 and p16 in the AAA+P7C3 group were significantly lower [SA-β-gal staining positive rate: (54.1±6.0)% vs. (74.1±4.4)%, p21/GAPDH: 0.38±0.02 vs. 0.61±0.07, p16/GAPDH: 0.50±0.13 vs. 0.77±0.03, all P < 0.05], but the expression of NAMPT was significantly increased (NAMPT/GAPDH: 1.25±0.28 vs. 0.88±0.07, P < 0.01). Conclusion:NAMPT agonist P7C3 can delay the senescence of VSMC and play a protective role in abdominal aortic aneurysm.

Investigation of serum vaspin, visfatin, chemerin and IL-18 levels in migraine patients / Investigação dos níveis séricos de vaspina, visfatina, quemerina e IL-18 em pacientes com migrânea

Abstract Background: Migraines are headaches caused by changes in the trigeminovascular metabolic pathway. Migraine headache attacks are associated with neurovascular inflammation, but their pathophysiological mechanisms have not been fully explained. Objective: To investigate the relationship between serum vaspin, visfatin, chemerin and interleukin-18 (IL-18) levels and the frequency of attacks in migraine headache. Methods: Three groups were established: migraine with aura (n = 50), migraine without aura (n = 50) and control group (n = 50). The migraine diagnosis was made in accordance with the International Classification of Headache Disorders-III beta diagnostic criteria. The analyses on serum vaspin, visfatin, chemerin and IL-18 levels were performed using the enzyme-linked immunosorbent assay method. Results: The serum vaspin, visfatin, chemerin and IL-18 levels were found to be significantly higher in the migraine patients than in the control group (p < 0.01). No statistically significant differences in serum vaspin, visfatin, chemerin and IL-18 levels were found among the migraine patients during attacks or in the interictal period (p>0.05). The serum visfatin and chemerin levels of the migraine patients were positively correlated with their serum IL-18 levels (p < 0.01), while their serum chemerin and visfatin levels were positively correlated with their serum vaspin levels (p < 0.05). Conclusions: This study showed that these biomarkers may be related to migraine pathogenesis. Nonetheless, we believe that more comprehensive studies are needed in order to further understand the role of vaspin, visfatin, chemerin and IL-18 levels in the pathophysiology of migraine headaches.

Resumo Introdução: A migrânea é causada por alterações nas vias metabólicas do sistema trigeminovascular. Crises de migrânea estão associadas à inflamação neurovascular, mas seus mecanismos patofisiológicos ainda não são totalmente explicados. Objetivo: Investigar a relação entre níveis séricos de vaspina, visfatina, quemerina e interleucina-18 (IL-18) e a frequência de crises de migrânea. Métodos: Três grupos foram formados: migrânea com aura (n = 50), migrânea sem aura (n = 50) e grupo controle (n = 50). A migrânea foi diagnosticada de acordo com os critérios da Classificação Internacional das Cefaleias (ICHD-III). As análises dos níveis séricos de vaspina, visfatina, quemerina e IL-18 foram realizadas utilizando-se o método imunoenzimático (ELISA). Resultados: Os níveis séricos de vaspina, visfatina, quemerina e interleucina-18 (IL-18) foram significativamente mais elevados em pacientes com migrânea do que no grupo controle (p < 0.01). Nenhuma diferença estatisticamente significativa foi observada nos níveis séricos de vaspina, visfatina, quemerina e interleucina-18 (IL-18) entre os pacientes com migrânea durante crises ou no período interictal (p>0,05). Os níveis séricos de visfatina e quemerina em pacientes com migrânea se correlacionaram positivamente com os níveis séricos de IL-18 (p < 0,01), ao passo que os níveis séricos de quemerina e visfatina se correlacionaram positivamente com os níveis séricos de vaspina (p < 0,05). Conclusões: Este estudo demonstrou que estes biomarcadores podem estar relacionados à patogênese da migrânea. Contudo, acreditamos que estudos mais abrangentes são necessários a fim de melhor compreendermos o papel dos níveis de vaspina, visfatina, quemerina e IL-18 na fisiopatologia da migrânea.

Association of visfatin with thyroid dysfunction in patients with nonalcoholic fatty liver disease / 临床肝胆病杂志

Recent studies have found that visfatin is involved in lipid metabolism in patients with nonalcoholic fatty liver disease (NAFLD), and about 25% of NAFLD patients have thyroid dysfunction. It has also been found that visfatin is involved in autoimmune regulation through the autocrine or paracrine pathways, which is associated with the involvement of the thyroid gland in autoimmune regulation of the body. This article preliminarily explores the association of visfatin with thyroid dysfunction in NAFLD patients and points out that the measurement of visfatin may help with the early identification of thyroid dysfunction in NAFLD patients.

Local brain hypothermia and nicotinamide phosphoribosyltransferase synergistically protect cerebral ischemia-reperfusion injury in rats / 国际脑血管病杂志

Objective To investigate the neuroprotective effect and its mechanism of local hypothermia combined with nicotinamide phosphoribosyltransferase (NAMPT) on cerebral ischemia-reperfusion injury in rats.Methods Fifty-four Sprague-Dawley rats were randomly divided into sham operation group,model group,NAMPT group,local hypothermia group,and combined treatment group (NAMPT + local hypothermia).A rat model of local cerebral ischemia-reperfusion was induced by suture method.Infarct volume and cerebral edema volume were assessed by 2,3,5-triphenyltetrazolium chloride staining after 2 h cerebral ischemia and 24 h reperfusion in rats.Evans blue staining was used to assess the extent of blood-brain barrier damage,and a 12-point scale was used to assess neurological deficits.Results The infarct volume in the local hypothermia group,NAMPT group,and combination treatment group was significantly lower than that in the model group (all P ＜0.05).The infarct volume in the combination treatment group was significantly lower than that in the NAMPT group (P ＜0.05).The infarct volume in the combination treatment group was lower than that in the local hypothermia group,but it did not reach statistical significance.The neurological function scores of the local hypothermia group,NAMPT group,and combination treatment group were significantly lower than those of the model group (all P ＜0.05).The score of the combined treatment group was significantly lower than that of the NAMPT group and the local hypothermia group (all P＜0.05).Evans blue leakage in the local hypothermia group,NAMPT group,and combination treatment group was lower than that in the model group (all P ＜0.05),but the differences between each treatment group were not statistically significant.Conclusion NAMPT and local hypothermia combination therapy showed better neuroprotective effects on cerebral ischemia-reperfusion injury,suggesting that the combination therapy had clinical transformation prospects.

The role of nicotinamide phosphoribosyltransferase in ischemic stroke / 国际脑血管病杂志

At present,there is still no effective drug treatment for acute ischemic stroke except intravenous thrombolysis.Recently,nicotinamide phosphonbosyltransferase (NAMPT) has been found to reduce neurological deficit after cerebral ischemia and reperfusion.This article reviews the neuroprotective effect of NAMPT in ischemic stroke and its potential mechanisms.

Serum visfatin levels in non-obese women with polycystic ovary syndrome and matched controls

OBJECTIVE: The purpose of the current study was to compare the circulating levels of visfatin between women with polycystic ovary syndrome (PCOS) and those without PCOS and to assess the correlations between visfatin levels and various parameters. METHODS: This case-control study recruited 74 PCOS patients and 74 age- and body mass index (BMI)-matched controls. Serum visfatin levels were evaluated using the enzyme-linked immunosorbent assay. Women with PCOS were divided into 2 subgroups based on the presence of clinical or biochemical hyperandrogenism. The possible differences in serum visfatin levels between the hyperandrogenic and non-hyperandrogenic groups were also assessed. RESULTS: Visfatin levels in PCOS patients were similar to those in the controls. However, hyperandrogenic patients had significantly higher mean serum visfatin levels than those in non-hyperandrogenic patients (3.87 ng/mL; 95% confidence intervals [CIs], 3.09–4.85 in hyperandrogenic group vs. 2.69 ng/mL; 95% CIs, 2.06–3.52 in non-hyperandrogenic group; P=0.038). In women with PCOS, visfatin levels positively correlated with BMI (r=0.23; P=0.047) and the log free androgen index (FAI) (r=0.27; P=0.021) and negatively correlated with high-density lipoprotein (HDL) cholesterol levels (r=−0.37; P=0.025). Except for HDL cholesterol levels, these correlations were also observed in controls. CONCLUSION: Visfatin levels in PCOS patients were similar to those in the controls. However, hyperandrogenic patients showed significantly higher serum visfatin levels than those of non-hyperandrogenic patients, and visfatin had a positive linear correlation with FAI in both PCOS patients and controls.

The serum level and significance of adiponectin, visfatin and resistin in patients with rheumatoid arthritis / 中华风湿病学杂志

Objective To determinate the serum level of adiponectin,visfatin and resistin in patients with rheumatoid arthritis (RA) and to explore its role in the development of RA and its clinical significance.Methods Blood samples were collected from 67 cases of RA patients and 65 healthy controls,the serum levels of adiponectin,visfatin and resistin were tested by enzyme linked immunosorbent assay (ELISA).Adipokine levels in different groups were compared using the Mann-Whitney U test.Spearman test and multivariate analysis were used to evaluate the correlation with clinical and laboratory parameters [erythrocyte sedimentation rate (ESR),C reactive protein (CRP),blood platelet (PLT),rheumatoid factor (RF),anti-cyclic citrullinated peptide (CCP),disease activity score (DAS)28].Results The serum level of adiponectin,visfatin and resistin in RA patients were significantly higher than those in healthy controls[8.17(4.51,28.53) μg/ml vs 6.83(4.48,25.32) μg/ml,U=1 663,P=0.019];[6.56(5.34,8.40) ng/L vs 4.24 (3.01,6.65) ng/L,U=762,P=0.001];[10.65 (5.99,20.70) ng/ml vs 6.12 (4.49,9.98) ng/ml,U=1 406.5,P=0.000].The serum level of visfatin was positively correlated with RF in patients with RA (r=0.186,P=0.013),and serum levels of adiponectin,visfatin and resistin were positively correlated with DAS28 (r=0.370,0.263,0.285;P＜0.05).The level of visfatin in RA patients with high activity group was higher than that in the low and moderate activity group [4.37(2.72,7.53)ng/L vs 4.11 (3.11,6.44) ng/L,U=133.5,P=0.019].Conclusion Multivariate analysis has showm that ESR,PLT and DAS28 can significantly affect adiponectin,and CRP can significantly influence resistin.Serum level of adiponectin,visfatin and resistin are elevated and correlate with DAS28,suggesting that they may be involved in the chronic inflammation of RA.

Expression of Nampt and bone morphogenetic protein 7 in spontaneous diabetic mice and effect of nicotinamide mononucleotide on fibrosis of rats' glomerular cells HBZY-1 in high glucose culture / 中国药理学与毒理学杂志

OBJECTIVE To investigate the mechanism of nicotinamide mononucleotide (NMN) on inflammation and fibrosis between endogenous nicotinamide phosphoribosyltransferase (NAMPT) and bone morphogenetic protein 7 (BMP-7) in diabetic glomerular cells.METHODS ① In vivo,spontaneous diabetic C57/BL6 mice and wild C57/BL6 mice were divided into two groups.When blood glucose was above (34.2±1.9) mmol· L-1,renal histology of diabetic mice became obvious.The protein expressions of Nampt and nuclear transcription factors-kappa B p65 (NF-κB p65),silent mating type information regulation 2 homolog 1 (SIRT1) and BMP7 were analyzed by lengths of immunofluorescence.② In vitro,rats' glomerular cells HBZY-1 were incubated with glucose 200 mmol· L-1 for different lengths of time (0,24,48 and 72 h) and at different concentrations of NMN (0,50,100 and 200 iμmol· L-1).The protein levels of Nampt and BMP7 were detected by Western blotting and the protein expressions of NF-κB p65 and α-SMA were measured by immunofluorescence assay.The protein levels of Nampt,BMP7 and NF-κB p65 were detected by Western blotting after HBZY-1 cells were treated with NMN 100 μmol· L-1 and FK866 10 μmol· L-1 for 24 h.RESULTS ① In vivo,the glomeruli of diabetic C57/BL6 mice showed obvious atrophy.Fluorescence intensity of Nampt was increased (P＜0.05),but that of BMP7 and SIRT1 in renal glomeruli cells was decreased compared with the wild type (P＜0.01).② In vitro,HBZY-1 cells were cultured in glucose 200 mmol· L-1 for 48 and 72 h.The protein expression of NAMPT was increased,but that of BMP7 was decreased (P＜0.05,P＜0.01).Expressions of NF-κB p65 and α-SMA were increased (P＜0.01) by immunofluorescence.The expression of BMP7 was increased after treatment with glucose 200 mmol· L-1,followed by NMN 50,100 and 200 μmol · L-1 for 24 h (P＜0.01).The expressions of NAMPT and NF-κB p65 were decreased (P＜0.01).The expressions of Nampt and NF-κB p65 in glucose 5.6 mmol· L1 +FK866 and glucose 5.6 mmol· L-1+ NMN groups were increased (P＜0.01),but the expression of BMP7 did not change.CONCLUSION Upregulation of endogenous Nampt obviously intervenes in BMP7 expression in the process of glomerular inflammatory fibrosis in severe diabetes.NMN can affect the protein expression of BMP7 via a special Nampt signaling pathway.

The predictive value of plasma visfatin and adiponectin in early atherosclerosis in patients with type 2 diabetes / 中华老年医学杂志

Objective To evaluate the association between plasma levels of visfatin and adiponectin with carotid arterial intima-media thickness in patients with type 2 diabetes.Methods A total of 100 elderly patients with type 2 diabetes and 30 healthy elderly controls were recruited from August 2015 to August 2016 at our hospital.Based on carotid arterial intima-media thickness(IMT) assessed via color Doppler ultrasonography,patients with diabetes were further divided into a thickened IMT group (40 cases)and a non-thickened IMT group (60 cases).Plasma levels of visfatin and adiponectin were analyzed and compared,and their associations with carotid arterial IMT were evaluated.Results Compared with the control group,patients in both the thickened IMT group and the non-thickened IMT group had significantly higher serum levels of visfatin(19.1 ± 2.8)μg/L vs.(40.6±3.9)μg/L,(28.9±3.6)μg/L,which was markedly higher in the thickened IMT group than in the non-thickened IMT group(P＜0.05).In addition,compared with the control group,patients in the thickened IMT group and the non-thickened IMT group had significantly lower serum levels of adiponectin(1 590±330)mg/L,(610±170)mg/L,(930±210)mg/L,which was markedly lower in the thickened IMT group than in the non-thickened IMT group (P ＜ 0.05).Furthermore,Pearson's analysis showed that the plasma level of visfatin was positively correlated with carotid arterial IMT(r =0.721,P＜0.05),and the plasma level of adiponectin was negatively correlated with carotid arterial IMT(r=-0.688,P＜0.05).Conclusions Plasma levels of visfatin and adiponectin are closely related to carotid arterial IMT in patients with type 2 diabetes,andmay be used as plasma markers for the prediction of early atherosclerosis.

Relationship between blood levels of visfatin and glycolipid metabolism and mild cognitive impairment in patients with type 2 diabetes / 中国医师杂志

Objective To explore the relationship between blood levels of visfatin and glycolipid metabolism and mild cognitive impairment (MCI) in patients with type 2 diabetes (T2DM).Methods From February 2016 to January 2017,a total of 91 patients with T2DM were recruited as investigation objects.Cognitive function was measured by Montreal Cognitive Assessment (MoCA),and fasting serum was collected to determine the relevant laboratory indexes.Results In the 91 patients,50 cases developed MCI.Compared to non-MCI group,MCI group had significant difference in age,total cholesterol,insulin,insulin resistance index,visfatin,MoCA score and diabetic retinopathy (P ＜ 0.05).Pearson correlation analysis showed that the MoCA score was negatively correlated with visfatin and insulin resistance index (P ＜0.05).Further logistic regression analysis showed that age,diabetic retinopathy,insulin resistance index and visfatin were independent risk factors for MCI in T2DM patients.Conclusions MCI in T2DM patients increases with the increasing of elder,diabetic retinopathy,insulin resistance index,and visfatin.

Regulation effect of endogenous nicotinamide phosphoribosyl transferase on Vimentin expression of glomerular cells in high concentration of glucose / 吉林大学学报(医学版)

Objective:To investigate the regulation effect of endogenous nicotinamide phosphoribosyl transferase (Nampt) on the Vimentin expression of glomerular cells in high concentration glucose,and to clarify the mechanism of formation of diabetic kidney inflammation fibrosis.Methods:The C57/BL6 diabetic mice were selected and the kidney tissues were collected,and the wild C57/L86 mice were used as control group;the pathological section and tissue fluorescence staining were performed.The expression and location of endogenous Nampt and Vimentin in the glomerular cells were detected by immuno-focused technology.The HBZY-1 cells were randomly divided into 4 groups:low concentration of glucose (LG,0.56 mmol · L-1) control group,high concentration glucose (HG,200 mmol · L-1) group,HG +-FK866 group and HG+nicotinamide mononucleotide (NMN) group.In HG group,the cells were treated with FK866 (10 μmol · L-1) and NMN (1 mmol · L-1) for 24 h after cultured with HG for 5 d.The expression levels of Nampt,Vimentin,nuclear factor-kappa B p65 (NF-κBp65) and sirtuin type 1 (Sirt1) were detected by immunofluorescence and Western blotting methods.The expression levels of Nampt and Vimentin were detected by RT-PCR and Western blotting methods.Results:The shape and size of glomerulus had obvious atrophy of the mice in severe diabetic group compared with normal C57/BL6 mice group.The expression level of Vimentin in glomerular cells was increased with the increasing of endogenous Nampt (P＜0.01).When the HBZY-1 cells were cultured in HG condition,the exprssion levels of Nampt,Vimentin and NF-kB p65 were obviously increased while the Sirt1 expression levels was significantly decreased compared with control group (P＜ 0.01).The expression levels of Nampt,Vimentin and NF-κB p65 in glomerular cells in HG+FK866 HG+ NMN groups were singnificartyly decreased compared with control group (P＜0.01).Conclusion:The endogenous Nampt over-expression in glomerular cells can enhance the expression of Vimentin under high concentration of glucose stress through NF-κBp65 and Sirt1 signal pathway.

Regulation effect of endogenous nicotinamide phosphoribosyl transferase on Vimentin expression of glomerular cells in high concentration of glucose / 吉林大学学报(医学版)

Objective:To investigate the regulation effect of endogenous nicotinamide phosphoribosyl transferase (Nampt) on the Vimentin expression of glomerular cells in high concentration glucose,and to clarify the mechanism of formation of diabetic kidney inflammation fibrosis.Methods:The C57/BL6 diabetic mice were selected and the kidney tissues were collected,and the wild C57/L86 mice were used as control group;the pathological section and tissue fluorescence staining were performed.The expression and location of endogenous Nampt and Vimentin in the glomerular cells were detected by immuno-focused technology.The HBZY-1 cells were randomly divided into 4 groups:low concentration of glucose (LG,0.56 mmol · L-1) control group,high concentration glucose (HG,200 mmol · L-1) group,HG +-FK866 group and HG+nicotinamide mononucleotide (NMN) group.In HG group,the cells were treated with FK866 (10 μmol · L-1) and NMN (1 mmol · L-1) for 24 h after cultured with HG for 5 d.The expression levels of Nampt,Vimentin,nuclear factor-kappa B p65 (NF-κBp65) and sirtuin type 1 (Sirt1) were detected by immunofluorescence and Western blotting methods.The expression levels of Nampt and Vimentin were detected by RT-PCR and Western blotting methods.Results:The shape and size of glomerulus had obvious atrophy of the mice in severe diabetic group compared with normal C57/BL6 mice group.The expression level of Vimentin in glomerular cells was increased with the increasing of endogenous Nampt (P＜0.01).When the HBZY-1 cells were cultured in HG condition,the exprssion levels of Nampt,Vimentin and NF-kB p65 were obviously increased while the Sirt1 expression levels was significantly decreased compared with control group (P＜ 0.01).The expression levels of Nampt,Vimentin and NF-κB p65 in glomerular cells in HG+FK866 HG+ NMN groups were singnificartyly decreased compared with control group (P＜0.01).Conclusion:The endogenous Nampt over-expression in glomerular cells can enhance the expression of Vimentin under high concentration of glucose stress through NF-κBp65 and Sirt1 signal pathway.

Inhibition of Nicotinamide Phosphoribosyltransferase Induces Apoptosis in Estrogen Receptor-Positive MCF-7 Breast Cancer Cells / 한국유방암학회지

PURPOSE: Tumor cells have increased turnover of nicotinamide adenine dinucleotide (NAD⁺), the main coenzyme in processes including adenosine diphosphate-ribosylation, deacetylation, and calcium mobilization. NAD⁺ is predominantly synthesized in human cells via the salvage pathway, with the first component being nicotinamide. Nicotinamide phosphoribosyltransferase (NAMPT) is the key enzyme in this pathway, and its chemical inhibition by FK866 has elicited antitumor effects in several preclinical models of solid and hematologic cancers. However, its efficacy in estrogen receptor (ER)-positive and human epidermal growth factor receptor 2-positive breast cancer cells has not been previously investigated. In this study, we aimed to deplete the NAD⁺ content of MCF-7 cells, a model cell line for ER-positive breast cancer, by inhibiting NAMPT in order to evaluate downstream effects on p53 and its acetylation, p21 and Bcl-2-associated X protein (BAX) expression, and finally, apoptosis in MCF-7 breast cancer cells. METHODS: MCF-7 cells were cultured and treated with FK866. NAD⁺ levels in cells were determined colorimetrically. Levels of p53 and its acetylated form were determined by Western blotting. Expression of p21 and BAX was determined by real-time polymerase chain reaction. Finally, levels of apoptosis were assessed by flow cytometry using markers for annexin V and propidium iodide. RESULTS: FK866 treatment was able to increase p53 levels and acetylation, upregulate BAX and p21 expression, and induce apoptosis in MCF-7 cells. Addition of exogenous NAD⁺ to cells reversed these effects, suggesting that FK866 exerted its effects by depleting NAD⁺ levels. CONCLUSION: Results showed that FK866 could effectively inhibit NAD⁺ biosynthesis and induce programmed cell death in MCF-7 cells, suggesting that NAMPT inhibitors may be useful for the treatment of ER-positive breast cancers.

Effect of curcumin on visfatin and zinc-α2-glycoprotein in a rat model of non-alcoholic fatty liver disease

ABSTRACT PURPOSE: To investigate the effect of curcumin on visfatin and zinc-α2-glycoprotein (ZAG) expression levels in rats with non-alcoholic fatty liver disease (NAFLD). METHODS: Fifty-six male rats were randomly divided into a control group (n=16) and model group (n=40) and were fed on a normal diet or a high-fat diet, respectively. Equal volumes of sodium carboxymethyl cellulose (CMC) were intragastrically administered to the control group for 4 weeks. At the end of the 12th week, visfatin and ZAG protein expression levels were examined by immunohistochemistry. Visfatin mRNA levels were measured by semi-quantitative reverse transcription polymerase chain reaction. RESULTS: Compared with the control group, the model group showed significantly increased expression of visfatin in liver tissue (P < 0.01) and significantly decreased expression of ZAG (P < 0.01). These effects were ameliorated by curcumin treatment. CONCLUSIONS: Visfatin and zinc-α2-glycoprotein may be involved in the pathogenesis of NAFLD. Treatment of NAFLD in rats by curcumin may be mediated by the decrease of visfatin and the increase of non-alcoholic fatty liver disease.

Effects of different degrees of intermittent hypoxia on NF-κB, IL-10 and visfatin in 3T3-L1 adipocytes / 天津医药

Objective To determine levels of nuclear factor (NF)-κB, interleukin (IL)-10, and visfatin in adipocytes treated by different degrees of intermittent hypoxia (IH), and to investigate the mechanism of IH leading to insulin resistance (IR). Methods The cell model of intermittent hypoxia/re-oxygenation (IH/ROX) in obstructive sleep apnea (OSA) was established. Differentiation mature 3T3-L1 adipocytes, were randomly divided into 10 groups including four different-frequency intermittent hypoxia groups(IH1-4, fixed intermittent hypoxia scheme for 1.5%O2 45 s and then re-oxygen 21%O2 for 2 min 15 s, 4 min 15 s, 5 min 45 s and 8 min 45 s, 60 times circulation), and their normal oxygen control groups (SC1-4, instead each IH group 1.5%O2 to 21%O2, the rest groups were treated as same as IH group), continuous hypoxia group (CH, 10%O2 for 6 h) and normal oxygen control group (CC, 21%O2 for 6 h). ELISA method was used to determine the levels of IL-10 and visfatin in the supematant of adipocytes. Western blot method was used to determine the protein levels of NF-κB p65 and visfatin. Real-time PCR method was used to determine the mRNA levels of IL-10 and visfatin. Results The protein and mRNA expressions of IL-10 were significantly lower in IH group and CH group than those of control groups (P<0.01). The levels of NF-κB p65 protein were significantly increased in IH group and CH group than those of control group. The protein and mRNA expressions of visfatin were significantly higher in IH1, IH2 and CH groups than those of control group (P<0.01). Conclusion As a prominent feature of OSA pathophysiology, IH may take part in insulin resistance of OSA patients by abnormally secreting NF-κB, IL-10 and visfatin in adipocytes.

Insights into the role of NAMPT and its inhibitors in the metabolism of tumor cells / 肿瘤

Different from normal cells, tumor cells display a metabolic profile by facilitate glycolysis pathway to provide adenosine triphosphate (ATP) quickly for cell growth and survival. Therefore, the demand of nicotinamide adenine dinucleotide (NAD), an important coenzyme in glycolysis pathway, increases obviously in tumor cells. It has been reported that tumor cells synthetize NAD through the primary salvage pathway, in which nicotinamide phosphoribosyltransferase (NAMPT) is the rate-limiting enzyme. As such, NAMPT regulates the cellular metabolism directly through controlling NAD synthesis and indirectly via affecting the activity of NAD-dependent enzymes. Meanwhile, NAMPT can ensure the viability of tumor cells by up-regulating the level of nicotinamide adenine dinucleotide phosphate (NADPH). Recently, NAMPT has been proposed as a new potential target for cancer treatment, and a number of studies have implicated that the NAMPT inhibitor has obvious anti-cancer effects. This review summarizes the alternation of metabolism in tumor cells, highlights the crucial role of NAMPT in cellular metabolism, and discusses the anticancer effect and the potential clinical application of NAMPT inhibitors in cancer treatment.

Visfatin effects on telomerase gene expression in AGS gastric cancer cell line.

AIM: The aim of this study was to assess visfatin expression and its effect on human telomerase gene expression in AGS gastric cancer cell line. MATERIALS AND METHODS: In this study, human gastric cancer (AGS) cell line was established as an in vitro model. Reverse transcription polymerase chain reaction (RT‑PCR) and enzyme‑linked immunosorbent assay was performed to show that visfatin expression in messenger ribonucleic acid (mRNA) and protein level respectively. After stimulating with increasing concentrations of visfatin for times of 24 h and 48 h, cell proliferation was assessed by 2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide (XTT) assay. In order to investigate telomerase gene expression affected by visfatin in AGS cell line, total RNA was extracted and complementary deoxyribonucleic acid was synthesized buy using commercially available kits. Expression of human telomerase reverse transcriptase (hTERT) mRNA was carried out by real‑time RT‑PCR. RESULTS: After visfatin treatment gastric cell line proliferation was enhanced and was increased the expression of hTERT. CONCLUSIONS: The obtained data showed that visfatin induces endogenously gastric cancer cell proliferation and increases telomerase (hTERT) gene expression, as a cancer gene. Based on this study, it is suggested that expression of this adipocytokine protein in real samples could be biomarker for gastric cancer.

Protective effect of phenylephrine on irradiated injury of osteoblasts in rats / 吉林大学学报(医学版)

Objective To investigate the effect of phenylephrine on the morphology and the expression level of Nampt mRNA in the irradiated osteoblasts of the rats,and to clarify the possible protective effects of phenylephrine on this injury.Methods The osteoblasts cultured by conventional tissue were randomly divided into control group, phenylephrine group,simple irradiation group,and phenylephrine+irradiation group.The cells in phenylephrine group were treated with 100 mmol· L-1 phenylephrine 30 min before irradiation.The cells in simple irradiation group were irradiated with 8 Gy X-ray.The osteoblasts were observed by microscope and were collected at the time points of 8,16,32 and 64 h after irradiation.The Nampt mRNA expression level in osteoblasts was examined by RT-PCR.Results There was no obviously morphological and number changes of osteoblasts in phenylephrine group compared with control group.The osteoblasts in simple irradiation group were atrophied,especially at 8 h;the number of osteoblasts in simple irradiation group was 0.82,0.37,0.24 and 0.21 times as control group respectively at 8,16,32 and 64 h. Atrophy of osteoblasts was alleviated in phenylephrine+irradiation group;at 8, 16,32 and 64 h,the number of osteoblasts was 0.91,0.83,0.72 and 0.75 times as control group,and was 1.09,2.24,3.00 and 3.60 times as simple irradiation group. The expression level of Nampt mRNA in the osteoblasts in simple irradiation group was reduced compared with control group (P < 0.01) at 8, 16, 32 and 64 h.There were significant differernces of the expression levels of Nampt mRNA in the osteoblasts between phenylephrine+irradiation group and control group (P < 0.01),and there were significant differernces between phenylephrine + irradiation group and simple irradiation group (P < 0.01 ) at the different time points. Conclusion Phenylephrine could reduce the atrophy of osteoblasts in the rats induced by irradiation and up-regulate the expression level of Nampt mRNA.Its protective effect on irradiated injury may be related to the up-regulation of the expression level of Nampt mRNA.

The expression of visfatin in liver of diabetic rats and it′s relationship with type 2 diabetic / 重庆医学

Objective To explore the effects of visfatin in glucose metabolism by testing the expression of visfatin in liver of rats in different glucose metabolic statuses .Methods SD rats were randomly divided into five groups :normal control group (NC group) ,diet induce obesity group (DIO group) ,diabetes mellitus group (DM group) ,diabetes controlled by insulin group (INS group)and diabetes controlled by metformin group (MET group) .Tested the data of blood glucose (FPG) ,triglyceride(TG) ,total cholesterol(TC) ,free fat acid(FFA) ,fasting insulin(Fins) .The liver of rats was used to test visfatin ,glucose‐6‐phosphatase(G‐6‐pase) mRNA by RT‐PCR and visfatin ,AMP‐activated protein kinase‐α (AMPKα) ,phosphor‐AMP‐activated protein kinase‐α (p‐AMPKα) protein by Western blot .Results FBG of group DM increased than group NC and DIO (P< 0 .01) ;FBG of group INS and MET decresed than group DM (P < 0 .01) ;HOMA‐IR of group DIO and DM increased than group NC (P < 0 .01) ;HOMA‐IR of group DM increased than group DIO (P< 0 .01) .ISI of group DIO and DM decresed than group NC (P< 0 .01) ;ISI of group DM de‐creased than group DIO(P< 0 .01) .TG of group DIO increased than group NC (P< 0 .01) .TG of group INS and MET decreased than group DM (P< 0 .05) .The level of TG and TC of group DM ,INS and MET increased than group NC (P< 0 .05) .The level of serum FFA of group DM ,INS and MET were significantly higher than group NC (P < 0 .05) ;FFA of group DM increased than group DIO(P< 0 .05) .The expression of visfatin mRNA of group DM increased than group NC and DIO (P< 0 .05) ;visfatin mRNA of group INS and MET decreased than group DM (P< 0 .01) .Group DM ,INS and MET had a significantly higher level of G‐6‐Pase mRNA of than group DIO( P < 0 .05) ;Group MET had a significantly lower level of G‐6‐Pase mRNA of than group DM (P <0 .05) .The expression of visfatin protein of group DM ,INS and MET increased than group NC ( P < 0 .05) .The expression of AMPKα protein of group DM ,INS and MET decresed than group NC(P< 0 .05) ;AMPKα of group DM decresed than group DIO (P<0 .05) .The expression of p‐AMPKα protein of group DIO ,DM ,INS and MET decresed than group NC (P< 0 .01) .Conclusion The ex‐pression of visfatin in liver of SD rats might have something to do with insulin resistance and diabetes .We could′t consider that visfatin can affect the pathway of metformin activated AMPK to decrease blood glucose .