Effect of omeprazole on antiplatelet activities of W1 / 国际药学研究杂志

Objective To investigate the antiplatelet effects of candidate drug W1 when combined with omeprazole,respec?tively.Methods The experimental rats were randomly divided into 5 groups:control group,clopidogrel(10 mg/kg)group,clopido?grel(10 mg/kg)+omeprazole(80 mg/kg)group,W1(3 mg/kg)group,and W1(3 mg/kg)+omeprazole(80 mg/kg)group.Four hours after oral administration of the drugs,the rats were measured for their platelet aggregation rate;Western blot was used to deter?mine the protein expressions of P2Y12 receptor,P-Akt and P-Erk.Results For the platelet aggregation rate,compared with the con?trol group,the 4 groups decreased significantly(P<0.01);the platelet aggregation rate in the clopidogrel + omeprazole group in?creased significantly than that in the clopidogrel group(42% and 20.4%,respectively,P<0.01);the platelet aggregoction rate (30.9%)in W1+omeprazole group was significantly higher(P<0.01)than that in the W1 group(20.5%),which was lower than that in the clopidogrel+omeprazole group(P<0.01).For the protein expression detected by the western blotting,there were no signif?icant changes in the expression of P2Y12 receptor on the platelet surface,in the clopidogrel or W1 group in comparison with the clopi?dogrel+omeprazole or W1+omeprazole group,respectively,while the phosphorylation level of Akt and Erk was up-regulated in the clopidogrel+omeprazole or W1+omeprazole group compared with the clopidogrel or W1 group,and the up-ragulatory effect of omepra?zole was weaker in the W1+omeprazole group than that in the clopidogrel+omeprazole group. Conclusion Combined use of omepra?zole could inhibit the antiplatelet activities of clopidogrel or W1,with the inhibitory effect weaker in W1 group than in clopidogrel group,suggesting that the risk for the combination of omeprazole with W1 is likely less than that for the combination with clopidogrel.

Effect of Atorvastatin on Expression of Peroxisome Proliferator-activated Receptor Beta/delta in Angiotensin II-induced Hypertrophic Myocardial Cells In Vitro / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To explore the effect of atorvastatin on cardiac hypertrophy and to determine the potential mechanism involved.</p><p><b>METHODS</b>An in vitro cardiomyocyte hypertrophy from neonatal rats was induced with angiotensin II (Ang II) stimulation. Before Ang II stimulation, the cultured rat cardiac myocytes were pretreated with atorvastatin at different concentrations (0.1, 1, and 10 μmol/L). The following parameters were evaluated: the myocyte surface area, 3H-leucine incorporation into myocytes, mRNA expressions of atrial natriuretic peptide, brain natriuretic peptide, matrix metalloproteinase 9, matrix metalloproteinase 2, and interleukin-1β, mRNA and protein expressions of the δ/β peroxisome proliferator-activated receptor (PPAR) subtypes.</p><p><b>RESULTS</b>It was shown that atorvastatin could ameliorate Ang II-induced neonatal cardiomyocyte hypertrophy in the area of cardiomyocytes, 3H-leucine incorporation, and the expression of atrial natriuretic peptide and brain natriuretic peptide markedly. Meanwhile, atorvastatin also inhibited the augmented mRNA level of several cytokines in hypertrophic myocytes. Furthermore, the down-regulated expression of PPAR- δ/β at both the mRNA and protein levels in hypertrophic myocytes could be significantly reversed by atorvastatin treatment.</p><p><b>CONCLUSIONS</b>Atorvastatin could improve Ang II-induced cardiac hypertrophy and inhibit the expression of cytokines. Such effect might be partly achieved through activation of the PPAR-δ/β pathway.</p>

mRNA and protein expression of leukotriene B4 receptor 2 during transition from colitis to colon cancer / 国际药学研究杂志

Objective: To observe mRNA and protein expression of leukotriene B4 (LTB4) receptor 2 (BLT2)in mice during the course of development from colitis to colitis-associated cancer. Methods: ICR mice were used to establish the animal model of colitis-associated colon cancer and randomly divided into control group and experimental group. We began to administer inducer on d0. Mice were sacrificed at 2, 3, 5, 7, 9, 13, and 18w, respectively. Histopathological changes in colons were examined. The protein and mRNA expression of BLT2 in colons were measured by immunohistochemical assay and real-time quantitative PCR, respectively. Results: Histological study showed that with the extension of time, the lesions of mice colons aggravated, first a mild inflammation, then atypical hyperplasia, and finally to colon cancer. Immunohistochemical staining showed that the expression of BLT2 protein was low in normal colon, but high in inflammatory lesions, especially in inflammatory cells. There was no BLT2 expression in atypical hyperplasia and cancer cells, while BLT2 was highly expressed in the stroma and lumans of cancer tissue. Compared with the control group, mRNA expression of BLT2 in the experimental group was significantly higher at 2, 13 and 18w (P<0.05); and very significantly higher at 3, 5, 7 and 9w(P<0.01). Conclusion: The mouse model of colitis-associated colon cancer was developed in this study. The expression of BLT2 changed in the course of colitis development, indicating that BLT2 may play an important role in the transition process from colitis to colon cancer.

Fusion of human orphan G protein-coupled receptors GPR45, GPR85 or GPR174 with Gi1α and their expression in insect Sf9 cells / 军事医学科学院院刊

Objective To obtain the fusion genes of several human orphan G protein coupled receptors (oGPCRs) with Gi1α subtype of G protein and their expression system. Methods The whole open reading frames of GPR45, GPR85, GPR174 and Gilα were cloned by RT-PCR from HepG2 cDNA separately,and the corresponding fusion genes were amplified by overlap extension PCR. Then, the fusion genes-containing pBacmids were successfully constructed with the Bac-to-Bac baculovirus expression system indicated by specific transposition and virus recombination. The insect Sf9 cells were transfected with pBacmid-oGPCRs-Gi1α, and the supernatant containing recombinant virus was harvested. With the supernatant, insect Sf9 cells were infected under an optimized condition (MOI=5, infection time=72 h) and the fusion proteins were prepared and detected by Western blotting.Results The three fusion genes of GPCR45, GPR85 or GPR174 with Gi1α were obtained. The corresponding fusion proteins could be properly prepared in Sf9 cells.Conclusion Human oGPCRs could be fused with Gilα, and the fusion genes could be expressed using the Bac-to-Bac baculovirus expression system in insect Sf9 cells.

Lymphocyte GRK2 expression of the very elderly with chronic heart failure / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the correlation of lymphocyte G protein-coupled receptor kinases 2 (GRK2) expression of the very elderly with chronic heart failure (HF) and heart ejection fraction (EF).</p><p><b>METHODS</b>16 elderly patients with chronic heart failure were divided into 2 groups as following: EF < 45% (n=7), EF > or = 45% (n=9); and health elderly as control (n=8). Lymphocytes were obtained from blood, reverse transcription polymerase chain reaction were used to measure GRK2 mRNA levels.</p><p><b>RESULTS</b>Lymphocyte GRK2 mRNA levels of EF < 45% group were higher than that of EF > 45% group, which were greater than that of control.</p><p><b>CONCLUSION</b>Elevation of lymphocyte GRK2 levels in HF is associated with heart EF, lymphocytes may provide a surrogate for monitoring cardiac GRK2 in human HF.</p>

Effect of metoprolol on the expression of GRK2 in lymphocyte of advanced elderly patients with chronic heart failure / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of metoprolol on the expression of G protein-coupled receptor kinases 2 (GRK2) in lymphocyte of advanced elderly patients with chronic heart failure.</p><p><b>METHODS</b>32 elderly patients with chronic heart failure were divided into control group and metoprolol group, 16 each. Conventional therapy was used in the control group, conventional therapy plua metoprolol was used in metoprolol group. The treatment courses were 8 weeks in both groups.</p><p><b>RESULTS</b>Left ventricular end-diastolic diameter and left ventricular ejection fraction were not different between the two groups. Lymphocyte GRK2 mRNA level in metoprolol group was lower than that in control group.</p><p><b>CONCLUSION</b>Metoprolol can inhibit the expression of GRK2 in lymphocyte of advanced elderly patients with chronic heart failure.</p>

Involvement of leukotrine B4 receptors in the inflammatory responses and immunological regulation in vitro / 中国应用生理学杂志

<p><b>AIM</b>BLT1 and BLT2 were both recently cloned and identified as two subtypes of leukotrine B4 (LTB4) receptors. With the usage of U-75302 and LY255283, the specific antagonists of BLT1 and BLT2 respectively, the involvement of BLT1 and BLT2 in the inflammatory and immunological responses was in vitro explored.</p><p><b>METHODS</b>(1) To investigate inhibition of U-75302 and LY255283 on the proliferation of rat synovial cells, 3H-TdR incorporation into the cells was quantified. (2) Flow cytometric assay for interferon-gamma (IFN-gamma) and interleukine 4 (IL-4) profiles in CD4+ T lymphocytes from rat spleen was carried out to determine the ratio of Th1/Th2.</p><p><b>RESULTS</b>(1) For inhibition on rat synovial cells proliferation, U-75302 exerted its effect only at a high concentration of 10 micromol/L and LY255283 at the concentrations of 10 micromol/L-10 micromol/L. (2) Both U-75302 and LY255283 could elevate the percentage of Th2, but could not influence that of Th1.</p><p><b>CONCLUSION</b>BLT1 and BLT2 were involved in the synovial cells proliferation change the ratio of Th1/Th2. Their meaning served as targets for prevention and treatment of infectious diseases should be emphasized.</p>

Subcellular localization and tissues expression profile of hGPCRc: an orphan G protein-coupled receptor / 生物工程学报

As a member of orphan G protein-coupled receptors (oGPCRs), hGPCRc was cloned from human colon tissue and analyzed by bioinformatic softwares. It was showed that the corresponding amino acids of hGPCRc formed seven-transmembrane domains as the key characteristic of GPCRs. Then, the recombinant GFP-hGPCRc was constructed by fussing hGPCRc into pEGFP-N1 carrying green fluorescent protein (GFP) gene, and CHO-K1 cells were subsequently transfected with the GFP-hGPCRc or pEGFP-N1. The green fluorescence protein expression in the two different transfected cells was observed under the laser scanning confocal microscopy (LSCM). It was showed that green fluorescence protein was distributed in the whole bodies of the cells transfected with pEGFP-N1, but mainly distributed on the plasma membrane and cytoplasm membrane transfected with GFP-hGPCRc. Thus, the localization on the membrane of hGPCRc was accorded with the predication by bioinformatic analysis. The expression analysis of hGPCRc by RT-PCR indicated that hGPCRc was abundantly expressed in heart, kidney, cerebel and colon etc., but absent in liver, cerebra, small intestine and muscle etc. The expressing profile of hGPCRc could provide some useful clues to understanding its effects on embryonic development and physiological functions.