Oral JS-38, a metabolite from Xenorhabdus sp., has both anti-tumor activity and the ability to elevate peripheral neutrophils / 中国天然药物

AIM@#JS-38 (mitothiolore), a synthetic version of a metabolite isolated from Xenorhabdus sp., was evaluated for its anti-tumor and white blood cell (WBC) elevating activities.@*METHOD@#These anti-proliferative activities were assessed in vitro using a panel of ten cell lines. The anti-tumor activities were tested in vivo using B16 allograft mouse models and xenograft models of A549 human lung carcinoma and QGY human hepatoma in nude mice. The anti-tumor interactions of JS-38 and cyclophosphamide (CTX) or 5-fluorouracil (5-Fu) were studied in a S180 sarcoma model in ICR mice. Specific stimulatory effects were determined on peripheral neutrophils in normal and CTX- and 5-Fu-induced neutropenic mice.@*RESULTS@#The IC50 values ranged from 0.1 to 2.0 μmol·L(-1). JS-38 (1 μmol·L(-1)) caused an increase in A549 tumor cell apoptosis. Multi-daily gavage of JS-38 (15, 30, and 60 mg·kg(-1)·d(-1)) inhibited in vivo tumor progression without a significant effect on body weight. JS-38 additively enhanced the in vivo anti-tumor effects of CTX or 5-Fu. JS-38 increased peripheral neutrophil counts and neutrophil rates in normal BALB/c mice almost as effectively as granulocyte colony-stimulating factor (G-CSF). In mice with neutropenia induced by CTX or 5-Fu, JS-38 rapidly restored neutrophil counts.@*CONCLUSION@#These results suggest that JS-38 has anti-tumor activity, and also has the ability to increase peripheral blood neutrophils.

Cilostazol inhibits insulin-stimulated expression of sterol regulatory binding protein-1c via inhibition of LXR and Sp1

Hepatic steatosis is common in obese individuals with hyperinsulinemia and is an important hepatic manifestation of metabolic syndrome. Sterol regulatory binding protein-1c (SREBP-1c) is a master regulator of lipogenic gene expression in the liver. Hyperinsulinemia induces transcription of SREBP-1c via activation of liver X receptor (LXR) and specificity protein 1 (Sp1). Cilostazol is an antiplatelet agent that prevents atherosclerosis and decreases serum triglyceride levels. However, little is known about the effects of cilostazol on hepatic lipogenesis. Here, we examined the role of cilostazol in the regulation of SREBP-1c transcription in the liver. The effects of cilostazol on the expression of SREBP-1c and its target genes in response to insulin or an LXR agonist (T0901317) were examined using real-time RT-PCR and western blot analysis on cultured hepatocytes. To investigate the effect of cilostazol on SREBP-1c at the transcriptional level, transient transfection reporter assays and electrophoretic mobility shift assays (EMSAs) were performed. Cilostazol inhibited insulin-induced and LXR-agonist-induced expression of SREBP-1c and its downstream targets, acetyl-CoA carboxylase and fatty acid synthase, in cultured hepatocytes. Cilostazol also inhibited activation of the SREBP-1c promoter by insulin, T0901317 and Sp1 in a luciferase reporter assay. EMSA analysis showed that cilostazol inhibits SREBP-1c expression by repressing the binding of LXR and Sp1 to the promoter region. These results indicate that cilostazol inhibits insulin-induced hepatic SREBP-1c expression via the inhibition of LXR and Sp1 activity and that cilostazol is a negative regulator of hepatic lipogenesis.

An evaluation of the mechanism of ABCA7 on cellular lipid release in ABCA7-HEC293 cell / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>ABCA7 is a member of the ABCA subfamily that shows a high degree of homology to ABCA1 and, like ABCA1, mediates cellular cholesterol and phospholipid release by apolipoproteins when transfected in vitro. However, expression of ABCA7 has been shown to be downregulated by increased cellular cholesterol while ABCA1 was upregulated.</p><p><b>METHODS</b>The underlying mechanism for this effect was examined in ABCA1 or ABCA7-transfected HEC293. Lipid content in the medium and cells was determined by enzymatic assays. Gene expression was quantitated by real time PCR, and protein content was determined by Western blotting.</p><p><b>RESULTS</b>While ABCA7 mRNA was decreased by 25-hydroxycholesterol treatment, ABCA1 was apparently increased. Treatment with the synthetic LXR agonist T0901317 (T09) upregulated ABCA1 expression and apoAI-mediated cellular lipid release in ABCA1-transfected HEC293 cells, but ABCA7 expression and cellular lipid release in ABCA7-transfected HEC293 cells showed no obvious changes.</p><p><b>CONCLUSION</b>The ABCA7 gene is regulated by sterol in a direction opposite to that of ABCA1.</p>

Effect of liver X receptor agonist T0901317 on endothelin-1 induced murine HL-1 cardiomyocytes hypertrophy / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the role of liver X receptors (LXRs) on endothelin-1 (ET-1) induced murine HL-1 cardiomyocytes hypertrophy.</p><p><b>METHODS</b>Cultured murine HL-1 cardiomyocytes were divided into four experiment groups: (1) CONTROL GROUP:treated with DMSO; (2) T0901317 group:treated with LXRs agonist T0901317 (1 µmol/L); (3) ET-1 group:treated with ET-1 (1 nmol/L); (4) T0901317 + ET-1 group:treated with T0901317 (1 µmol/L) for 8 hours, then treated with ET-1 (1 nmol/L). Twenty-four hours later, immunofluorescent staining was performed on HL-1 cells, the surface area of HL-1 cells was analyzed with NIH Image J software, and the synthetic rate of protein in HL-1 cells was detected by (3)H-leucine incorporation. The mRNA level of atrial natriuretic peptide (ANP) and β-myosin heavy chain (β-MyHC) was measured by quantitative realtime PCR. The effect of T0901317 on mRNA expression of ANP was also detected after LXRs gene silencing.</p><p><b>RESULTS</b>The surface area of HL-1 cells, mRNA expression of ANP and β-MyHC, and (3)H-leucine incorporation in ET-1 group were 2.00 ± 0.29, 1.98 ± 0.47, 2.13 ± 0.39 and 1.79 ± 0.17, respectively, which were significantly higher than those of control group (1.00 ± 0.26, 1.00 ± 0.21, 1.00 ± 0.31 and 1.00 ± 0.03, respectively, all P < 0.05). Compared with ET-1 group, the surface area of HL-1 cells, mRNA expression of ANP and β-MyHC, and (3)H-leucine incorporation were significantly decreased in T0901317 + ET-1 group (1.24 ± 0.25, 1.19 ± 0.21, 1.48 ± 0.27 and 1.15 ± 0.11, respectively, all P < 0.05). After inhibition of LXRα/β expression in HL-1 cardiomyocytes using the specific siRNAs, the mRNA expression of ANP in T0901317 + ET-1 group was 1.78 ± 0.05, which was similar as that in ET-1 group (1.94 ± 0.17, P > 0.05).</p><p><b>CONCLUSION</b>T0901317, an agonist of LXRs, could inhibit ET-1 induced cardiac hypertrophy in vitro, and LXR ligand-mediated inhibition on ANP mRNA expression by T0901317 is receptor dependent.</p>

Beneficial effects of liver X receptor agonist on adipose-derived mesenchymal stem cells transplantation in mice with myocardial infarction / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the effects of liver X receptor (LXR) agonist on adipose-derived mesenchymal stem cells (AD-MSCs) implantation into infarcted hearts of mice.</p><p><b>METHOD</b>AD-MSC(Fluc+) which stably expressed firefly luciferase (Fluc) were isolated from β-actin-Fluc transgenic mice and characterized by flow cytometry. Male FVB mice were randomly allocated into the following four groups (n = 10 each): (1) sham group; (2) MI + PBS group; (3) MI + AD-MSC(Fluc+) group; (4) MI + AD-MSC(Fluc+) + LXR agonist (T0901317) group. AD-MSC(Fluc+) or PBS were injected intramyocardial into peri-infarcted region of mice heart after permanent left anterior descending (LAD) artery ligation. Bioluminescence imaging (BLI) was performed for quantification of injected cells retention and survival. Cardiac function was evaluated by echocardiography.</p><p><b>RESULTS</b>The AD-MSC(Fluc+) were positive for CD44 and CD90 by flow cytometry. BLI evidenced the firefly luciferase expression of AD-MSC(Fluc+) which was positively correlated with cell numbers (r(2) = 0.98). The results of BLI in vivo revealed that LXR agonist could improve the survival of AD-MSC(Fluc+) at day 7, 14 and 21 after transplantation compared with AD-MSC(Fluc+) alone group. Cardiac function was further improved in combination therapy group compared with AD-MSC(Fluc+) alone group (P < 0.05).</p><p><b>CONCLUSIONS</b>LXR agonist T0901317 can improve the retention and survival of intramyocardial injected AD-MSC(Fluc+) post-MI, and the combination therapy of T0901317 and AD-MSC(Fluc+) has a synergetic effect on improving cardiac function in this model.</p>

Degradation products of different water content sevoflurane in carbon dioxide absorbents by gas chromatography-mass spectrometry analysis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Sevoflurane is currently used as a volatile inhalation anesthetic with many clinical advantages. A representative degradation product, compound A, was quantitatively measured to investigate whether there are different reactions between two kinds of water content sevoflurane formulations with different carbon dioxide (CO2) absorbents.</p><p><b>METHODS</b>A closed-circle breathe bag with the Dräger Fabius GS anesthesia apparatus was used as an artificial rubber lung. The experiments were grouped according to different sevoflurane formulations: group A: higher-water sevoflurane (Ultane); group B: lower-water sevoflurane (Sevoness). During the experiment, CO2 (200 ml/min) was continually perfused to keep the end-tidal pressure of CO2 (P(ET)CO2) at 35 - 45 mmHg. The artificial ventilation was set to 6 L/min, and the breathing rate at 12 breaths/min. The circuit was operated with constant fresh gas flow rate (1 L/min) and the sevoflurane concentration was kept at 1.0 minimum alveolar concentration (MAC) for 240 minutes. At 0, 10, 20, 30, 60, 90, 120, 180 and 240 minutes, gas was collected from the Y-piece. Gas chromatography/mass spectrometry (GC/MS) was used to quantify the major degradation product, compound A, with different water content sevoflurane. PETCO2 and sevoflurane concentration, and the temperature of the canister were continuously monitored during the experiment.</p><p><b>RESULTS</b>There were no significant differences in P(ET)CO2 and sevoflurane concentrations between the two groups. Drägersorb 800 plus produced the highest concentrations of compound A compared with other sodalimes, and Sevoness in Drägersorb 800 plus generated more compound A than Ultane (P < 0.05). There were significant differences in the peak and average compound A concentrations between Ultane and Sevoness with Drägersorb 800 plus (P < 0.05), while the compound A concentration produced by Sodasorb grase and sofonolime in the two groups showed no significant difference (P > 0.05). In the same group, the peak and average of compound A concentration produced by Sodasorb grase and sofonolime showed significant difference with Drägersorb 800 plus (P < 0.05).</p><p><b>CONCLUSION</b>The water content of sevoflurane and potassium hydroxide in CO2 absorbent can influence compound A production.</p>

Liver X receptor agonist T0901317 inhibits TGF-β1-induced α-SMA expression in normal human lung fibroblasts / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of liver X receptor agonist T0901317 on transforming growth factor-β1 (TGF-β1)-induced expression of α-smooth muscle actin (α-SMA) in normal human lung fibroblasts.</p><p><b>METHODS</b>Primary normal human lung fibroblast isolated from the lung specimens of lung cancer patients by explant culture technique were identified with immunostaining for vimentin and keratin. The cells in passages 4 to 10 were treated with T0901317 and/or TGF-β1, and RT-PCR, Western blotting and immunofluorescence assay were used to detect α-SMA expression in the fibroblasts.</p><p><b>RESULTS</b>Lung fibroblast expressed vimentin but not keratin. The results of RT-PCR, Western blotting and immunofluorescence assay all showed that normal human lung fibroblasts constitutively expressed α-SMA under baseline condition, and TGF-β1 at 5 ng/ml induced a significant upregulation of α-SMA both at the mRNA and protein levels. Liver X receptor agonist T0901317 (5 µg/ml) significantly inhibited TGF-β1-induced upregulation of α-SMA expression.</p><p><b>CONCLUSION</b>Liver X receptor agonist T0901317 can inhibit the upregulation of α-SMA in normal human lung fibroblasts induced by TGF-β1, suggesting the potential value of liver X receptor agonist in the treatment of lung fibrosis.</p>

A comparison between vital capacity induction and tidal breathing induction techniques for the induction of anesthesia and compound A production / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Vital capacity induction and tidal breathing induction are currently administered for inhalation induction of anesthesia with sevoflurane. The aim of this study was to compare them using sevoflurane with respect to induction time, complications of inhalation induction, and compound A production in adult patients.</p><p><b>METHODS</b>Fifty-one women with American Society of Anesthesiologists physical status I-II undergoing mammary gland tumorectomy were randomly assigned to receive either vital capacity induction or tidal breathing induction with 8% sevoflurane at 6 L/min followed by laryngeal mask airway insertion. Induction times, complications of inhalation induction, and vital signs were recorded. Inspired concentrations of compound A were assayed and sofnolime temperatures were monitored at one-minute intervals after sevoflurane administration.</p><p><b>RESULTS</b>The time to loss of eyelash reflex was significantly shorter with the vital capacity induction technique than with the tidal breathing induction technique ((43.8 ± 13.4) seconds vs. (70.8 ± 16.4) seconds, respectively; P < 0.01). Cardiovascular stability was similar in both groups. The incidence of complications was significantly less with the vital capacity induction technique than with the tidal breathing induction technique (7.7% vs. 32%, respectively; P < 0.01). However, the mean and maximum concentrations of compound A during induction were significantly higher in the vital capacity group than those in the tidal breathing group (P < 0.05); compound A concentration at the beginning of anesthesia maintenance was (40.73 ± 10.83) ppm in the vital capacity group and (29.45 ± 7.51) ppm in tidal breathing group (P = 0.019).</p><p><b>CONCLUSION</b>For inhalation induction of anesthesia, the vital capacity induction was faster and produced fewer complications than that for tidal breathing induction, but increased compound A production in the circuit system.</p>

Effect of liver X receptors agonists T0901317 on expression of FAT/CD36 gene mRNA in adult human skeletal muscle cell / 中国应用生理学杂志

<p><b>AIM</b>To investigate the liver X receptors agonists T0901317's effect on expression of FAT/CD36 gene mRNA in adult human skeletal muscle cell.</p><p><b>METHODS</b>Myotubes from humans were exposed to different T0901317 concentrations (0, 0.5, and 1.0 micromol/L) for 24 hours before experiments were performed. Then the expression of FAT/CD36 mRNA in skeletal muscle cell of each experimental group was detected by SYBR Green I real-time quantitative polymerase chain reaction. The relative data were compared among groups by 2-delta delta Ct method.</p><p><b>RESULTS</b>(1) The Ct mean of control group, T0901317 (0.5 micromol/L) group, T0901317 (1 micromol/L) group were analyzed and there was significant difference (P < 0.01). (2) The expression of FAT/CD36 mRNA with liver X receptors agonists T0901317 in human skeletal muscle cell in the T0901317 (0.5 micromol/L) group and T0901317 (1 micromol/L) group were 2.91 times and 3.03 times than the control group.</p><p><b>CONCLUSION</b>The expression of FAT/CD36 mRNA in human skeletal muscle cell afer the treatment of liver X receptors agonists T0901317 is increased, so we may propose that T0901317 may increase the risk of resistance in adult human skeletal muscle.</p>

Liver X receptor alpha inhibits LPS-induced inflammation by down-regulating IFR3 and GRIP1 in mouse Kupffer cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the possible mechanism of the inhibitory effect of liver X receptor alpha (LXRalpha) on lipopolysaccharide (LPS)-induced inflammation in mouse Kupffer cells (KCs).</p><p><b>METHODS</b>The KCs isolated from the liver of male KM mice and cultured in RPMI 1640 containing 20% FBS for 24 h were divided into control, LPS, T0901317, and LPS+T0901317 groups with corresponding treatments. The expressions of LXRalpha, interferon regulatory factor 3 (IRF3) and glucocorticoid receptor interacting protein 1 (GRIP1) in the KCs were detected by Western blotting. The levels of interferon beta (IFNbeta), tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) in the supernatant were detected by enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The level of LXRalpha protein was highest in T0901317 group and lowest in LPS group, and was significantly higher in LPS+T0901317 group than in LPS group but lower than in T0901317 group (P<0.05). The levels of IRF3 and GRIP1 protein were the highest in LPS group, and significantly lowered by T0901317 treatment (P<0.05). The expression of IRF3 and GRIP1 proteins in LPS group and LPS+ T0901317 group were significantly higher than those in the control and T0901317 groups (P<0.05). The concentration of IFN-beta was significantly higher in LPS group than in the control and T0901317 group (P<0.05), and decreased in LPS+T0901317 group in comparison with that in LPS group (P<0.05). IFN-beta was the lowest in T0901317 group. The levels of TNF-alpha and IL-1beta were the highest in LPS group (P<0.05), and comparable between the other 3 groups (P>0.05).</p><p><b>CONCLUSION</b>Pre-treatment with T0901317 before LPS stimulation can suppress the expressions of IRF3 and GRIP1 to inhibit the inflammation and hence Kupffer cell activation.</p>

Role of liver X receptors on cardiomyocyte hypertrophy / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the expression of liver X receptors (LXR) in hypertrophic myocardium and the effect of LXR agonist T0901317 on angiotensin II (AngII) induced cardiomyocyte hypertrophy.</p><p><b>METHODS</b>Transverse aortic coarctation (TAC) or sham operation were performed in 2-month-old wide type mice (C57/B6). Two weeks later, the expression of LXR in myocardium was detected by quantitative real-time PCR analysis and Western blot analysis. The effect of LXR agonist T0901317 on AngII-induced hypertrophy in cultured neonatal rat cardiomyocytes was also assessed.</p><p><b>RESULTS</b>Quantitative real-time PCR analysis and Western blot analysis showed that LXRalpha but not LXRbeta expression was upregulated post TAC both at mRNA and protein levels (All P < 0.05). AngII induced increased [(3)H] leucine incorporation and cardiomyocyte hypertrophy were significantly reduced by T0901317 in a dose-dependent manner (P < 0.05). T0901317 also dose-dependently inhibited atrial natriuretic peptide (ANP) gene expression in cardiomyocytes (P < 0.05).</p><p><b>CONCLUSION</b>Our findings strongly suggest that LXR is a potent mediator of cardiomyocyte hypertrophy and LXR activation could attenuate AngII induced cardiomyocyte hypertrophy in vitro.</p>

zVAD-fmk, unlike BocD-fmk, does not inhibit caspase-6 acting on 14-3-3/Bad pathway in apoptosis of p815 mastocytoma cells

In a preliminary study, we found that benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone (zVAD- fmk), unlike Boc-aspartyl(OMe)-fluoromethylketone (BocD-fmk), at usual dosage could not prevent genistein-induced apoptosis of p815 mastocytoma cells. This study was undertaken to reveal the mechanism underlying the incapability of zVAD-fmk in preventing this type of apoptosis. We observed that 14-3-3 protein level was reduced in genistein-treated cells and that BocD-fmk but not zVAD-fmk prevented the reduction of 14-3-3 protein level and the release of Bad from 14-3-3. We also demonstrated that truncated Bad to Bcl-xL interaction in genistein- treated cells was prevented by BocD-fmk but not by zVAD-fmk treatment. Our data indicate that BocD- fmk, compared to zVAD-fmk, has a certain preference for inhibiting 14-3-3/Bad signalling pathway. We also elucidated that this differential efficacy of BocD-fmk and zVAD-fmk resulted from the different effect in inhibiting caspase-6 and that co-treatment of zVAD-fmk and caspase-6 specific inhibitor substantially prevented genistein-induced apoptosis. Our data shows that caspase-6 plays a role on Bad/14-3-3 pathway in genistein-induced apoptosis of p815 cells, and that the usual dose of zVAD-fmk, in contrast to BocD-fmk, did not prevent caspase-6 acting on 14-3-3/Bad-mediated event.

Hidrofluoralcano como propelente dos aerossóis pressurizados de dose medida: histórico, deposição pulmonar, farmacocinética, eficácia e segurança / Hydrofluoroalkane as a propellant for pressurized metered-dose inhalers: history, pulmonary deposition, pharmacokinetics, efficacy and safety

OBJETIVO: Rever a literatura sobre o hidrofluoralcano como propelente dos inaladores de dose medida contendo medicamentos empregados na asma. FONTES DOS DADOS: O levantamento bibliográfico foi realizado em bancos de dados eletrônicos - MEDLINE, MDConsult, HighWire, Medscape e LILACS - e por pesquisa direta - referentes aos últimos 15 anos -, utilizando-se as seguintes palavras-chaves: hidrofluoralcano, asma e infância. SíNTESE DOS DADOS: Foram selecionados 43 artigos originais abordando a questão da substituição do clorofluorcarbono pelo hidrofluoralcano. Este gás mostrou-se como uma alternativa de propelente segura, com deposição pulmonar de 50 a 60 por cento e eficácia significativa quando comparado com placebo (p < 0,003) através de estudos clínicos controlados. A maioria das pesquisas realizadas com o hidrofluoralcano emprega o dipropionato de beclometasona. O custo anual aproximado do tratamento com dipropionato de beclometasona/hidrofluoralcano foi menor do que com dipropionato de beclometasona/clorofluorcarbono. Alguns estudos avaliaram o salbutamol, a fluticasona, a flunisolida e a associação fluticasona-salmeterol tendo o hidrofluoralcano como propelente em inaladores pressurizados de dose medida. CONCLUSÕES: Ficou evidenciada em adultos a eficácia e segurança do hidrofluoralcano como propelente de broncodilatadores e glicocorticosteróides em aerossol, bem como a melhor deposição pulmonar das partículas, de um modo geral. Todavia, são escassos os dados na literatura acerca do uso do hidrofluoralcano na faixa etária pediátrica. Seria importante conduzir estudos adicionais em crianças e adolescentes.