小檗碱对肾性高血压心肌肥厚模型大鼠左心室重塑的影响

The purpose of this study is to evaluate the effects and the underline mechanisms of berberine on the cardiac function and left ventricular remodeling in rats with renovascular hypertension. The renovascular hypertensive model was established by the two-kidney, two-clip （2K2C） method in Sprague-Dawley （SD） rats. Two weeks after surgery, all the operated SD rats were randomly assigned into four groups： （1） renovascular hypertensive model group; （2） berberine 5 mg · kg^-1 group; （3） berberine 10 mg · kg^-1 group ; （4） captopril 45 mg · kg^-1 group ; and the sham operated rats were used as control. Four weeks after the drugs were administered, the cardiac function was assessed. The ratios of heart weight to body weight （HW/BW）, left ventricular weight to body weight （LVW/BW） and right ventricular weight to body weight （RVW/BW） were compared between groups. Coronal sections of the left ventricular tissue （LV） were prepared for paraffin sections, picrosirius red and HE staining was performed. The left ventricular wall thickness （LVWT）, interventricular septal thickness （IVST）, the parameters of myocardial fibrosis indicated by interstitial collagen volume fraction （ICVF） and perivascular collagen area （ PVCA ） were assessed. Nitric oxide （ NO ）, adenosine cyclophosphate （ cAMP ） and guanosine cyclophosphate （cGMP） concentrations of left ventricular tissue were measured. Berberine 5 mg· kg^-1 and 10 mg · kg^-1 increased the left ventricular± dp/dt max. and HR. Berberine 10 mg · kg^-1 decreased HW/BW and LVW/BW. The image analysis showed that both 5 and 10 mg · kg^-1 of berberine decreased LVWT, ICVF and PVCA, while increased the NO and cAMP contents in left ventricular tissue. Berberine could improve cardiac contractility of 2K2C model rats, and inhibit left ventricular remodeling especially myocardial fibrosis in renovascular hypertension rats. And such effects may partially associate with the increased NO and cAMP content in left ventricular tissue.     

GJA1-20k attenuates Ang II-induced pathological cardiac hypertrophy by regulating gap junction formation and mitochondrial function

Cardiac hypertrophy(CH)is characterized by an increase in cardiomyocyte size,and is the most common cause of cardiac-related sudden death.A decrease in gap junction(GJ)coupling and mitochondrial dysfunction are important features of CH,but the mechanisms of decreased coupling and energy impairment are poorly understood.It has been reported that GJA1-20k has a strong tropism for mitochondria and is required for the trafficking of connexin 43(Cx43)to cell–cell borders.In this study,we investigated the effects of GJA1-20k on Cx43 GJ coupling and mitochondrial function in the pathogenesis of CH.We performed hematoxylin–eosin(HE)and Masson staining,and observed significant CH in 18-week-old male spontaneously hypertensive rats(SHRs)compared to age-matched normotensive Wistar-Kyoto(WKY)rats.In cardiomyocytes from SHRs,the levels of Cx43 at the intercalated disc(ID)and the expression of GJA1-20k were significantly reduced,whereas JAK-STAT signaling was activated.Furthermore,the SHR rats displayed suppressed mitochondrial GJA1-20k and mitochondrial biogenesis.Administration of valsartan(10 mg·kg^?1.d^?1,i.g.,for 8 weeks)prevented all of these changes.In neonatal rat cardiomyocytes(NRCMs),overexpression of GJA1-20k attenuated Ang II-induced cardiomyocyte hypertrophy and caused elevated levels of GJ coupling at the cell–cell borders.Pretreatment of NRCMs with the Jak2 inhibitor AG490(10μM)blocked Ang II-induced reduction in GJA1-20k expression and Cx43 gap junction formation;knockdown of Jak2 in NRCMs significantly lessened Ang II-induced cardiomyocyte hypertrophy and normalized GJA1-20k expression and Cx43 gap junction formation.Overexpression of GJA1-20k improved mitochondrial membrane potential and respiration and lowered ROS production in Ang II-induced cardiomyocyte hypertrophy.These results demonstrate the importance of GJA1-20k in regulating gap junction formation and mitochondrial function in Ang II-induced cardiomyocyte hypertrophy,thus providing a novel therapeutic strategy for patients with cardiomyocyte hypertrophy.     

The peroxisome proliferator-activated receptor-α （PPAR-α） agonist,AVE8134, attenuates the progression of heart failure and increases survival in rats

Aim： To investigate the efficacy of the peroxisome proliferator-activated receptor-α （PPARa） agonist, AVE8134, in cellular and experimental models of cardiac dysfunction and heart failure.
Methods： In Sprague Dawley rats with permanent ligation of the left coronary artery （post-MI）, AVE8134 was compared to the PPARy agonist rosiglitazone and in a second study to the ACE inhibitor ramipril. In DOCA-salt sensitive rats, efficacy of AVE8134 on cardiac hypertrophy and fibrosis was investigated. Finally, AVE8134 was administered to old spontaneously hypertensive rats （SHR） at a nonblood pressure lowering dose with survival as endpoint. In cellular models, we studied AVE8134 on hypertrophy in rat cardiomyocytes nitric oxide signaling in human endothelial cells （HUVEC） and LDL-uptake in human MonoMac-6 cells.
Results： In post-MI rats, AVE8134 dose-dependently improved cardiac output, myocardial contractility and relaxation and reduced lung and left ventricular weight and fibrosis. In contrast, rosiglitazone exacerbated cardiac dysfunction. Treatment at AVE8134 decreased plasma proBNP and arginine and increased plasma citrulline and urinary NOx/creatinine ratio. In DOCA rats, AVE8134 prevented development of high blood pressure, myocardial hypertrophy and cardiac fibrosis, and ameliorated endothelial dysfunction. Compound treatment increased cardiac protein expression and phosphorylation of eNOS. In old SHR, treatment with a low dose of AVE8134 improved cardiac and vascular function and increased life expectancy without lowering blood pressure. AVE8134 reduced phenylephrine-induced hypertrophy in adult rat cardiomyocytes. In HUVEC, Ser-1177-eNOS phosphorylation but not eNOS expression was increased. In monocytes, AVE8134 increased the expression of CD36 and the macrophage scavenger receptor 1, resulting in enhanced uptake of oxidized LDL.
Conclusion： The PPARa agonist AVE8134 prevents post-MI myocardial hypertrophy, fibrosis and cardiac dysfunction. AVE8134 has beneficial effects against hy     

去窦弓神经大鼠肥厚心室和主动脉的血管紧张素Ⅱ和AT1受体水平

AIM: Angiotensin II and AT1 receptor are the major effector components of renin-angiotensin system (RAS), andalso the main growth-stimulating factors in cardiovascular system. The present study was to observe these twofactors in the hypertrophied ventricles and aortas of sinoaortic-denervated rats. METHODS: Rats were examinedat 2, 10, and 16 weeks after sinoaortic denervation (SAD). The hypertrophy was evaluated by the ratio of organweight to body weight. Angiotensin II concentration and AT1 receptor mRNA expression were measured byradioimmunoassay and RT-PCR respectively, using a positive control of candesartan treatment. RESULTS: Aortichypertrophy existed in 2-, 10-, and 16-week SAD rats, left ventricular hypertrophy in 10- and 16-week SAD rats,and right ventricular hypertrophy in 16-week SAD rats. In all three kinds of examined SAD rats, plasma angiotensinII levels remained unchanged, indicating circulating RAS is at normal level in the chronic phase of SAD. However,cardiovascular tissue RAS was activated, as evidenced by increase of aortic angiotensin II concentrations at 10 and16 weeks after SAD, and up-regulation of aortic and left ventricular AT1 receptor mRNA expressions at 16 weeksafter SAD. CONCLUSION: The activated tissue RAS is secondary to the hypertrophy, and probably involved inthe maintenance of cardiovascular hypertrophy following SAD.     

In vitro anticancer property of a novel thalidomide analogue through inhibition of NF-κB activation in HL-60 cells

Aim： To investigate the anticancer property and possible mechanism of action of a novel sugar-substituted thalidomide derivative （STA-35） on HL-60 cells in vitro.
Methods： TNF-α-induced NF-κB activation was determined using a reporter gene assay. The MTT assay was used to measure cytotoxicity of the compound. The appearance of apoptotic Sub-G1 cells was detected by flow cytometry analysis. PARP cleavage and protein expression of NF-κB p65 and its inhibitor IκB were viewed by Western blotting.
Results： STA-35 （1-20 μmol/L） suppressed TNF-α-induced NF-κB activation in transfected cells （HEK293/pNiFty-SEAP） in a dose- （1-20 μmol/L） and time-dependent （0-48 h） manner. It was also shown that STA-35 exerted a dose-dependent inhibitory effect on HL-60 cell proliferation with an IC50 value of 9.05 lamol/L. In addition, STA-35 induced apoptosis in HL-60 cells, as indicated by the appearance of a Sub-G1 peak in the cell cycle distribution, as well as poly ADP-ribose polymerase （PARP） cleavage. Subsequently, both NF-κB p65 and its inhibitor IκB gradually accumulated in cytoplasmic extracts in a dose- and time-dependent manner, indicating the blockage of NF-κB translocation induced by TNF-α from the cytoplasm to the nucleus.
Conclusion： A novel sugar-substituted thalidomide derivative, STA-35, is potent toward HL-60 cells in vitro and induces apoptosis by the suppression of NF-κB activation.     

Puerarin blocks Na~+ current in rat ventricular myocytes

AIM: To study the effect of puerarin (Pue) on Na~+ channel in rat ventricular myocytes. METHODS: Whole-cell patch-clamp technique was applied on isolated cardiomyocytes from rats. RESULTS: Pue inhibited cardiac I_(Na) in a positive rate-dependent and dose-dependent manner, with an IC_(50) of 349 μmol/L. The kinetics of blockage of cardiac sodium channel by Pue resembled the ClassIa/Ic of antiarrhythmic agents. Pue 300 μmol/L did not alter the shape of the I-V curve of I_(Na), but markedly shifted the steady-state inactivation curve of I_(Na) towards more negative potential by 15.9 mV, and postponed the recovery of I_(Na) inactivation state from (21.9±1.6) ms to (54.4±3.4) ms (P<0.01 ). It demonstrated that the steady state of inactivation was affected by Pue significantly. CONCLUSION: Pue protected ventricular myocytes against cardiac damage and arrhythmias by inhibiting recovery from inactivation of cardiac Na~+ channels.     

在患有急性肺损伤的小猪的肺泡巨噬细胞中一氧化氮，表面活性剂和糖皮质激素对核因子-κB和激活蛋白-1的活性的调控作用

AIM： To investigate whether acute lung injury (ALI) in ventilated piglets with bacterial infection affects NF-κB and AP-1 expression in alveolar macrophages (AM) and whether nitric oxide (NO), surfactant (Surf), glucocorticoids (GC) affect NF-κB and AP-1 activation in AM in vivo and in vitro. METHODS: The animals were intraperitoneally injected Escherichia coli, which caused ALI. Nuclear extracts of AM were analyzed by electrophoretic mobility shift assay (EMSA) for the nuclear factor-kappa B (NF-κB) and activation protein-1 (AP-1) expression. Detection of IκB-α protein was from cytoplasmic extract by Western blotting. Immunocytochemistry staining was used for intracellular location of p65 subunits of NF-κB. RESULTS: In ex vivo experiments, strikingly higher expression of NF-κB and AP-1 by EMSA was found 6h after bacterial injection in contrast to the Normal group. In the NO, SNO,and GC groups, markedly attenuated NF-κB and AP-1 activation was observed. The NF-κB and AP-1 activation in Surf group showed lower levels of the expression. Immunoblotting of AM cytoplasmic extract showed low expression of IκB-α protein in the Control and Surf groups. The stronger expression was observed in the NO, GC,and SNO groups. AM of the Control and Surf groups showed intense nuclear staining, with decreased nuclear staining in the NO, GC and SNO groups. In in vitro experiment, it caused a significant increase in NF-κB and AP1 activity in AM 1h after exposure to lipopolysaccharides (LPS). In AM treated by LPS SNP and LPS GC, all showed decrease of DNA binding activity of NF-κB and AP-1 compared to those exposed to LPS Surf. Immunoblotting of AM cytoplasmic extract showed that LPS stimulation of AM resulted in the low expression of IκB-α protein, which was not observed in the presence of SNP and methylprednisolone. However, the surfact antdid not show such effect. LPS Surf-exposed AM had intense nuclear staining, whereas decreased nuclear staining in the LPS NO and LPS GC-treated cultures was found, confirming a decrease in NF-rd3 activity. CONCLUSION:Activation of NF-κB was found in AM of ventilated piglets with bacterial ALI. NO and GS could prevent NF-κB and AP-1 activation in vivo and in vitro. Surfactant has limited effects on NF-κB and AP-1 activity.     

Sodium ferulate inhibits cardiomyocyte hypertrophy induced by Ang Ⅱ through enhancement of NO/cGMP signaling pathway

OBJECTIVE The present study is to investigate the effect of SF on neonatal rat cardiomyocytes of myocardial hypertrophy induced by 0.1 μmol · L~(-1) angiotensinⅡ(AngⅡ) and explore the underlying mechanism.METHODS The cultured cardiomyocytes from Sprague Dawley neonate rats were randomly divided into: normal,model(AngⅡ0.1 μmol·L~(-1)), L-arginine(1000 μmol·L~(-1))group and SF(50, 100, 200 μmol·L~(-1)) group. To observe whether SF had nonspecific injurious effect on the cells,SF 200 μmol·L~(-1)was added into the normal cardiomyocytes. To determine whether the effect of SF on cardiomyocyte hypertrophy is associated with NO release, another two groups[NG-nitro-L-arginine-methyl ester(L-NAME)]1500 μmol·L~(-1) combined with SF 200 μmol·L~(-1) and L-arginine 1000 μmol·L~(-1)) were established. RESULTS After administration, Ang Ⅱ decreased the NO content, NOS and e NOS activity in supernatant of cultured cardiomyocytes, and decreased the content of c GMP and increased the content of c AMP in cardiomyocytes, up-regulation the expression of PKC-β, Raf-1, ERK1/2 and down-regulated the expression of MKP-1 and e NOS. SF 200 μmol·L~(-1) and L-argininesignificantly ameliorated these changes.SF ameliorate the cardiomyocyte hypertrophy which can be inhibited by L-NAME. These results indicate that SF can inhibit cardiomyocyte hypertrophy induced by AngⅡand the probable mechanism involved to promote NO/c GMP signaling pathway and inhibit PKC and MAPK signaling pathway. CONCLUSION These results indicate that SF can inhibit cardiomyocyte hypertrophy induced by AngⅡ and the probable mechanism involved to promote NO/c GMP signaling pathway and inhibit PKC and MAPK signaling pathway.     

Targeting at SUR2B/Kir6.1 subtype of ATP-sensitive potassium channel opener natakalim improves pressure overload-induced heart failure

Objective To explore the new stratigies targeting at SUR2B/Kir6.1 subtype against pressure overload-induced heart failure.Methods Pressure overload-induced heart failure was induced in Wistar rat by abdominal aortic banding(AAB).The effects of natakalim(1,3,9 mg·kg-1·d-1,10 weeks)were assessed on myocardial hypertrophy and heart failure,cardiac histology,vasoactive compounds,and gene expression.Isolated working heart and isolated tail artery helical strips were used to examine the influence of natakalim on heart and resistant vessels.Results Ten weeks after the onset of pressure overload,natakalim therapy potently inhibited cardiac hypertrophy and prevented heart failure.Natakalim inhibited the changes of left ventricular haemodynamic parameters,reversed the increase of heart mass index,left ventricular weight index and lung weight index remarkably.Histological examination demonstrated that there were no significant hypertrophy and fibrosis in hearts of pressure overload rat treated with natakalim.Ultrastructural examination of heart revealed well-organized myofibrils with mitochondria grouped along the periphery of longitudinally oriented fibers in natakalim group rats.The content of serum NO and plasma PGI2 was increased,while that of plasma ET-1 and cardiac tissue hydroxyproline,ANP and BNP mRNA was down-regulated in natakalim-treated rats.Natakalim at concentrations ranging from 0.01-100 μM had no effects on isolated working heart derived from Wistar rats;however,natakalim had endothelium-dependent vasodilation effects on the isolated tail artery helical strips precontracted with NE.Conclusions These results indicate that natakalim improves heart failure due to pressure overload by activating KATP channel SUR2B/Kir6.1 subtype and reversing endothelial dysfunction.     

Role of a new-type polypeptide in regulation of oxidative stress in cardiac hypertrophy through ox-CaMK Ⅱ

OBJECTIVE Oxidative stress is the imbalance between the production and removal of reactive oxygen species(ROS), leading to cel and tissue damage.There is growing evidence that excessive ROS is induced in cardiac hypertrophy and heart failure. Over accumulation of ROS subsequently activates ROS-sensitive downstream signaling pathways associated with pathological myocardial hypertrophy. Angiotensin Ⅱ(Ang Ⅱ)increases ROS in ventricular myocardium by acting on oxidized CaMK Ⅱ(ox-CaMK Ⅱ). This study will explore the role of a new polypeptide targeting on CaMKⅡ in the regulation of oxidative stress in cardiac hypertrophy through ox-CaMKⅡ, and provide a new target and a new way for the treatment of cardiac hypertrophy. METHODS H9c2 cells were divided into three groups. The control group was given complete medium. The cardiac hypertrophy group was administered by 100 nmol·L~(-1) AngⅡ. After 24 h of administration of Ang Ⅱ, the polypeptide group was given a new type polypeptide, 5 μg· m L~(-1). Oxidative stress was estimated in vivo by measuring ROS level,ox-CaMK Ⅱ expression and superoxide dismutase(SOD)activity. RESULTS Compared with that of the control group, the SOD activity in the cardiac hypertrophy group was decreased, while the new polypeptide could significantly improve the SOD activity. Among the three groups, the expression of ox-CaMK Ⅱ in the cardiac hypertrophy group was the highest, demonstrating that this novel polypeptide could reduce ox-CaMKⅡ expression. CONCLUSION In conclusion, our newly designed CaMKⅡ-targeted polypeptide can inhibit ROS-mediated downstream pathway by inhibiting ox-CaMK Ⅱ expression and reducing excessive ROS accumulation, thereby reversing cardiac hypertrophy.     

Regulation of activity of nuclear factor-kB and activator protein-1 by nitric oxide, surfactant and glucocorticoids in alveolar macrophages from piglets with acute lung injury

AIM: To investigate whether acute lung injury (ALl) in ventilated piglets with bacterial infection affects NF-κB and AP-1 expression in alveolar macrophages (AM) and whether nitric oxide (NO), surfactant (Surf), glucocorticoids (GC) affect NF-κB and AP-1 activation in AM in vivo and in vitro. METHODS: The animals were intraperitoneally injected Escherichia coli, which caused ALl. Nuclear extracts of AM were analyzed by electrophoretic mobility shift assay (EMSA) for the nuclear factor-kappa B (NF-κB) and activation protein-1 (AP-1) expression. Detection of IκB-α protein was from cytoplasmic extract by Western blotting. Immunocytochemistry staining was used for intracellular location of p65 subunits of NF-κB. RESULTS: In ex vivo experiments, strikingly higher expression of NF-κB and AP-1 by EMSA was found 6 h after bacterial injection in contrast to the Normal group. In the NO, SNO, and GC groups, markedly attenuated NF-κB and AP-1 activation was observed. The NF-κB and AP-1 activation in Surf g     

Gastric ulcer healing effect of wild honey and its combination with Turmeric（Curcuma domestica Val.） Rhizome on male Wistar rats

Gastric ulcer is a common disorder in human at any ages. In this research, the antiulcer activity of wild honey produced by Apis dorsata, alone or in combination with Turmeric Rhizome, was evaluated in healing acute gastric ulcer. Male Wistar albino rats（150-250 g） were induced ulcers with aspirin at 405 mg/kg BW and ethanol. Antiulcer evaluation was done based on the gastric acidity, numbers and diameter of ulcers, ulcer index, healing ratio, histological examinations, and body weight. The results showed that the groups given honey alone, turmeric alone, and combination of turmeric-honey displayed significant ulcer healing compared to the control group. Ulcers in the group administered with combination of turmeric-wild honey was different significantly from the turmeric alone and wild honey alone groups with increased body weight in that group. The result showed that wild honey（2125 mg/kg BW） had the greatest activity in healing ulcers among other groups. The combination of turmeric-wild honey had a good activity in healing ulcers and increased the body weight of the group.     

Inhibitory effect of 1,8-cineol （eucalyptol） on Egr-1 expression in lipopolysaccharide-stimulated THP-1 cells

Aim： To study the effects of 1,8-cineol （eucalyptol） on the expression of early growth response factor-1 （Egr-1） and NF-κB in the human monocyte THP-1 cell line stimulated by lipopolysaccharide （LPS）. Methods： The THP-1 cells were incu- bated with serial doses of 1,8-cineol （1, 10, and 100 mg/L, 30 min） before being stimulated with LPS （1 mg/L, 30 min）. The localization of Egr-1 in the THP-1 cells was detected by immunofluorescence and a laser scanning confocal microscope. The expression of Egr-1 in the nuclei and whole cell, and NF-κB in the nuclei, were measured by Western blot analysis. Results： When stimulated by LPS, the FITC- labeled Egr-1 was detected mainly in the nuclei. Moreover, the expression of Egr-1 in the whole cell increased markedly compared with the control cells. 1,8- Cineol pretreatment decreased the expression of Egr-1 in both the nuclei and whole cell of the LPS-stimulated THP- 1 cells, and this effect was concentration- dependent, but there was no reaction on the expression of NF-κB in the nuclei protein in the LPS-stimulated THP-1 cells. Conclusion： In a concentration-depen- dent manner, 1,8-Cineol reduces LPS-induced Egr-1 expression in nuclei and in whole cell of THP-1 cells, but shows no effect on NF-κB expression.     

Isoindolone derivative QSN-IOc induces leukemic cell apoptosis and suppresses angiogenesis via PI3K/AKT signaling pathway inhibition

Aim： 2-（4,6-Dimethoxy-l,3-dioxoisoindolin-2-yl） ethyl 2-chloroacetate （QSN-IOc） is one of isoindolone derivatives with antiproliferative activity against human umbilical vein endothelial cells （HUVECs）. The aim of this study was to investigate its antitumor activity in vitro and anti-angiogenic effects in vitro and in vivo. Methods： K562 leukemic cells and HUVECs were used for in vitro studies. Cell viability was examined using MTT assay. Cell apoptosis and mitochondrial transmembrane potential （Δψm） were detected with flow cytometry. Tube formation and migration of HUVECs were studied using two-dimensional Matrigel assay and wound-healing migration assay, respectively. VEGF levels were analyzed with RT-PCR and Western blotting. A zebrafish embryo model was used for in vivo anti-angiogenic studies. The molecular mechanisms for apoptosis in K562 cells and antiangiogenesis were measured with Western blotting. Results： In antitumor activity studies, QSN-IOc suppressed the viability of K562 cells and induced apoptosis in dose- and time- dependent manners. Furthermore, QSN-IOc dose-dependently decreased the Δψm in K562 cells, increased the release of cytochrome c and the level of Bax, and decreased the level of Bcl-2, suggesting that QSN-10c-induced apoptosis of K562 cells was mediated via the mitochondrial apoptotic pathway. In anti-angiogenic activity studies, QSN-IOc suppressed the viability of HUVECs and induced apoptosis in dose dependent manners. QSN-IOc treatment did not alter the Δψm in HUVECs, but dose-dependently inhibited the expression of VEGF, inhibited the tube formation and cell migration in vitro, and significantly suppressed the number of ISVs in zebrafish embryos in vivo. Furthermore, QSN-IOc dose-dependently suppressed the phosphorylation of AKT and GSK313 in both HUVECs and K562 cells. Conclusion： QSN-IOc is a novel antitumor compound that exerts both antitumor and anti-angiogenic effects via inhibiting the PI3K/ AKT/GSK313 signaling pathway.     

Effect of α-pinene on nuclear translocation of NF-κB in THP-1 cells

AIM: To study the effects of α-pinene on nuclear translocation of nuclear factor-κB (NF-κB) and the expression of the inhibitor of NF-κB (IκBα) in human monocyte THP-1 cell line. METHODS: THP-1 cells were incubated with α-pinene (1, 10, and 100 mg/L, for 30 min) before being stimulated with lipopolysaccharide (LPS, 1 mg/L, 30 min).The location of NF-κB p65 subunit (NF-κB/p65) in THP-1 cells was detected by immunofluorescence and laser scanning confocal microscope (LSCM). The expression of NF-κB/p65 in nuclei and that of IκBα in cytoplasm were measured by Western-blot analysis. RESULTS: The majority of FITC-labelled NF-κB/p65 was located in the nuclei being stimulated with LPS. Whereas, no such fluorescence was seen in the nuclei of the groups pretreated with α-pinene or control cells, α-Pinene pretreatment decreased the NF-κB/p65 nuclear translocation in LPS-stimulated THP-1 cells, and this effect was dose-dependent, but there was no reaction in LPS-unstimulated THP-1 cells, α-Pinene pretreatment increased IκBα protein level in cytoplasm, compared with that in LPS-stimulated THP-1 cells. CONCLUSION: In a dose-related fashion, α-pinene inhibits the nuclear translocation of NF-κB induced by LPS in THP- 1 cells, and this effect is partly due to the upregulation of IκBα expression.     

Therapeutic effects of DZ2002, a reversible SAHH inhibitor,on lupus-prone NZBxNZW F1 mice via interference with TLR-mediated APC response

Aim： To examine the therapeutic effects and underlying mechanisms of DZ2002, a reversible S-adenosyI-L-homocysteine hydrolase （SAHH） inhibitor, on lupus-prone female NZBxNZW F1 （NZB/W F1） mice. Methods： Female NZB/W F1 mice were treated orally with DZ2002 （0.5 mg.kgl-d-1） for 11 weeks, and the proteinuria level and body weight were monitored. After the mice ware euthanized, serum biochemical parameters and renal damage were determined. Splenocytes of NZB/W F1 mice were isolated for ex vivo study. Toll-like receptor （TLR）-stimulated human peripheral blood mononuclear cells （PBMCs） or murine bone marrow-derived dendritic cells （BMDCs） were used for in vitro study. Results： Treatment of the mice with DZ2002 significantly attenuated the progression of glomerulonephritis and improved the overall health. The improvement was accompanied by decreased levels of nephritogenic anti-dsDNA IgG2a and IgG3 antibodies, serum IL-17, IL-23p19 and TGF-13. In ex vivo studies, treatment of the mice with DZ2002 suppressed the development of pathogenic Th17 cells, significantly decreased IL-17, TGF-13, IL-6, and IL-23p19 production and impeded activation of the STAT3 protein and JNK/NF-KB signaling in splenocytes. DZ2002 （500 pmol/L） significantly suppressed TLR agonists-stimulated up-regulation in IL-6, IL-12p40, TNF-a, and IgG and IgM secretion as well as in HLA-DR and CD40 expression of dendritic cells among human PBMCs in vitro. DZ2002 （100 pmol/L） also significantly suppressed TLR agonists-stimulated up-regulation in IL-6 and IL-23p19 production in murine BMDCs, and prevented Thl7 differentiation and suppressed IL-17 secretion by the T cells in a BMDC-T cell co-culture system. Conclusion： DZ2002 effectively ameliorates lupus syndrome in NZB/W F1 mice by regulating TLR signaling-mediated antigen presenting cell （APC） responses.     

Vam3, a resveratrol dimer,inhibits cigarette smoke- induced cell apoptosis in lungs by improving mitochondrial function

Aim： To investigate the effects of Vam3 （a resveratrol dimer extracted from Vitis amurensis Rupr） on cigarette smoke （CS）-induced cell apoptosis in lungs in vitro and in vivo and the underlying mechanisms of action.
Methods： Human bronchial epithelial cell line BEAS-2B was exposed to cigarette smoke condensate （CSC, 300 mg/L）, and cell apoptosis was determined using flow cytometry and Hoechst staining. Mitochondrial membrane potential was examined with TMRE staining. ROS and ceramide levels were detected with DCFH-DA fluorescence and HPLC-MS/MS, respectively. Cytochrome c release was detected using immunofluorescence. Caspase-9 and neutral sphingomyelinase 2 expression was measured with Western blotting. The breast carcinoma cell line MCF7 stably expressing GFP-tagged Bax was used to elucidate the role of mitochondria in CS-induced apoptosis. For in vivo study, male mice were exposed to CS for 5 min twice a day for 4 weeks. The mice were orally administered Vam3 （50 mg·kg^-1·d^-1） or resveratrol （30 mg·kg^-1·d^-1） each day 1 h before the first CS exposure.
Results： Pretreatment of BEAS-2B cells with Vam3 （5 μmol/L） or resveratrol （5 μmol/L） significantly suppressed CSC-induced apoptosis, and prevented CSC-induced Bax level increase in the mitochondria, mitochondrial membrane potential loss, cytochrome c release and caspase-9 activation. Furthermore, pretreatment of BEAS-2B cells with Vam3 or resveratrol significantly suppressed CSC-stimulated intracellular ceramide production, and CSC-induced upregulation of neutral sphingomyelinase 2, the enzyme responsible for ceramide production in bronchial epithelial cells. Similar results were obtained in C6-pyridinium ceramide-induced apoptosis of GFP-Bax-stable MCF7 cells in vitro, and in the lungs of CS-exposed mice that were treated with oral administration of Vam3 or resveratrol.
Conclusion： Vam3 protects bronchial epithelial cells from CS-induced apoptosis in vitro and in vivo by preventing mitochondrial dysfunction.     

The anti-tumor properties of two tumstatin peptide fragments in human gastric carcinoma

Aim： The aim was to study the anti-tumor activities and mechanisms of two synthetic peptide fragments of tumstatin （alpha3 （iV） NCl domain） in human gastric carcinoma cells in vitro and in vivo. Methods： MTr assay and cell cycle assay were used to study the anti-tumor and anti-angiogenic activities of two peptide fragments in vitro. Apoptosis induced by the two peptide fragments was demonstrated by TUNEL assay and morphological observation. The orthotopic tumor model was established to investigate the activities of two peptide fragments in vivo. Intratumor vascularization and the expressions of VEGF, bFGF, Fas, FasL, Bax, Bcl-2, and caspase 3 were determined using immunohistochemistry and Western blot analysis. Results： Peptide 19 inhibited SGC-7901 proliferation and induced apoptosis both in vitro and in vivo. Notably, peptide 21 suppressed the proliferation of HUVEC-12 cells in vitro. Each peptide arrested both cell lines at the G0/G1 phase of the cell cycle, and they also synergistically suppressed in vitro and in vivo tumor growth. Immunohistochemistry and Western blot analysis revealed the strong expression of Fas, FasL and caspase 3 in orthotopic tumor tissues treated with peptide 19 alone or in combination with peptide 21. Decreased expressions of VEGF and bFGF and decreased microvessel density （MVD） in orthotopic tumor tissues were seen in mice treated with peptide 21 alone or in combination with peptide 19. Conclusion： Two tumstatin peptide fragments facilitate two unique antitumor activities. Thus, they are drug candidates in the treatment of gastric carcinoma.