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In addition to the essential pharmacologi-cal effects of opioids,situational cues associated with drug addiction memory are key triggers for drug seeking.CircRNAs,an emerging hotspot regulator in crown genet-ics-play an important role in central nervous system-relat-ed diseases.However,the internal mediating mechanism of circRNA in the field of drug reward and addiction mem-ory remains unknown.Here,we trained mice on a condi-tional place preference(CPP)model and collected nucle-us accumbens(NAc)tissues from day 1(T0)and day 8(T1)for high-throughput RNA sequencing.qRT-PCR revealed that circTmeff-1 was highly expressed in the NAc core but not in the NAc shell,suggesting that it plays a role in addiction memory formation.Meanwhile,the reverse regulation of circTmeff-1 by adeno-associated viruses could both inhibit the formation of addiction mem-ory in the NAc core or shell.Subsequently,the GO and KEGG analyses indicated 21 that circTmeff-1 might regu-late the addiction memory via the MAPK and AMPK path-ways.These findings suggest that circTmeff-1 in NAc plays a crucial role in morphine-dependent memory for-mation.
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Overexpressing of ATP-binding cassette (ABC) transporters is the essential cause of multidrug resistance (MDR), which is a significant hurdle to the success of chemotherapy in many cancers. Therefore, inhibiting the activity of ABC transporters may be a logical approach to circumvent MDR. Olmutinib is an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI), which has been approved in South Korea for advanced EGFR T790M-positive non-small cell lung cancer (NSCLC). Here, we found that olmutinib significantly increased the sensitivity of chemotherapy drug in ABCG2-overexpressing cells. Furthermore, olmutinib could also increase the retention of doxorubicin (DOX) and rhodamine 123 (Rho 123) in ABC transporter subfamily G member 2 (ABCG2)-overexpressing cells. In addition, olmutinib was found to stimulate ATPase activity and inhibit photolabeling of ABCG2 with [I]-iodoarylazidoprazosin (IAAP). However, olmutinib neither altered ABCG2 expression at protein and mRNA levels nor blocked EGFR, Her-2 downstream signaling of AKT and ERK. Importantly, olmutinib enhanced the efficacy of topotecan on the inhibition of S1-MI-80 cell xenograft growth. All the results suggest that olmutinib reverses ABCG2-mediated MDR by binding to ATP bind site of ABCG2 and increasing intracellular chemotherapeutic drug accumulation. Our findings encouraged to further clinical investigation on combination therapy of olmutinib with conventional chemotherapeutic drugs in ABCG2-overexpressing cancer patients.
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Objective To assess image quality of adaptive statistical iterative reconstruction (ASIR),conventional modelbased iterative reconstruction (MBIRc) and a new lung-specific setting (MBIRRP20 and MBIRNR40) from the new version of model-based iterative reconstruction (MBIRn) in submillisievert chest CT comparing with ASIR in standard-dose.Methods Two chest CT examinations were performed with standard-dose and low-dose in 30 patients.Low-dose CT images were reconstructed with ASIR,MBIRc and MBIRn,while standard-dose CT images were reconstructed with ASIR only.Objective image noise and SNR were measured on the same part from the back muscle and subcutaneous fat which located at the level of thoracic entry,trachea carina and hepatic portal.Image quality of lung,mediastinum and upper abdomen structures were evaluated on a 5-point scale.The results were compared with one-way ANOVA and Wilcoxon signed-rank tests.Results The effective dose equivalent for standard-dose CT was (3.01 ± 1.89) mSv,compared with (0.88 ± 0.83) mSv for low dose CT,which decreased by 70.76％.The mean image noise for low-dose MBIRNR40 was significantly lower than that of conventional-dose ASIR,low-dose ASIR and MBIRc (P＜0.05).The mean SNR for low-dose MRIRNR40 was significant ly higher than that of conventional dose ASIR,low-dose ASIR and MBIRc (P＜0.05).The subjective image noise score was significantly lower than that of ASIR and MBIRc,and the score of sharpness of details of the structures score for low dose MBIRn was significantly better than that of the ASIR and MBIRc (P＜0.05).Conclusion MBIRNR40 can significantly reduce image noise and improve SNR compared to ASIR and MBIRc in low-dose,even better than ASIR in standard dose,which reduce radiation dose by about 70％.In low-dose,MBIRPP20 can well display lung structures,and MBIRNR40 can display mediastinal and the upper abdominal structures.
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Objective To assess image quality of adaptive statistical iterative reconstruction (ASIR),conventional modelbased iterative reconstruction (MBIRc) and a new lung-specific setting (MBIRRP20 and MBIRNR40) from the new version of model-based iterative reconstruction (MBIRn) in submillisievert chest CT comparing with ASIR in standard-dose.Methods Two chest CT examinations were performed with standard-dose and low-dose in 30 patients.Low-dose CT images were reconstructed with ASIR,MBIRc and MBIRn,while standard-dose CT images were reconstructed with ASIR only.Objective image noise and SNR were measured on the same part from the back muscle and subcutaneous fat which located at the level of thoracic entry,trachea carina and hepatic portal.Image quality of lung,mediastinum and upper abdomen structures were evaluated on a 5-point scale.The results were compared with one-way ANOVA and Wilcoxon signed-rank tests.Results The effective dose equivalent for standard-dose CT was (3.01 ± 1.89) mSv,compared with (0.88 ± 0.83) mSv for low dose CT,which decreased by 70.76％.The mean image noise for low-dose MBIRNR40 was significantly lower than that of conventional-dose ASIR,low-dose ASIR and MBIRc (P＜0.05).The mean SNR for low-dose MRIRNR40 was significant ly higher than that of conventional dose ASIR,low-dose ASIR and MBIRc (P＜0.05).The subjective image noise score was significantly lower than that of ASIR and MBIRc,and the score of sharpness of details of the structures score for low dose MBIRn was significantly better than that of the ASIR and MBIRc (P＜0.05).Conclusion MBIRNR40 can significantly reduce image noise and improve SNR compared to ASIR and MBIRc in low-dose,even better than ASIR in standard dose,which reduce radiation dose by about 70％.In low-dose,MBIRPP20 can well display lung structures,and MBIRNR40 can display mediastinal and the upper abdominal structures.
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Objective To explore the risk factors of youth cerebral infarction and its correlation with clinical TOAST types.Methods 82 young patients with acute cerebral infarction(aged from 18 to 45 years old)were select-ed.The risk factors for youth cerebral infarction patients,and the proportion of TOAST subtype and related risk factors were analyzed.Results Risk factors for youth cerebral infarction were as follows:hyperlipidemia (χ2 =48.703,P <0.05),hypertension (χ2 =40.829,P <0.05),carotid sclerosis (χ2 =46.217,P <0.05),hyperhomocysteinemia (χ2 =40.255,P <0.05),smoking history (χ2 =7.853,P <0.05),diabetes (χ2 =18.256,P <0.05)and family history (χ2 =5.944,P <0.05),heart disease (χ2 =5.754,P <0.05).The proportion of their TOAST subtypes were as following:small artery occlusion lacunar(SAD)(39.0%),large artery atherosclerosis(LAA)(24.4%),stroke of other undetemined etiology(SUE)(17.0%),and acute stroke of other detemined etiology(SOE)(12.2%),cardio-embolism(CE)type(7.3%).Major risk factors for LAA subtype included hyperlipidemia,hypertension and carotid atherosclerosis;Major risk factor for SUE subtype was hyperhomocysteinemia;Major risk factors for SOE included hypertension,diabetes.Major risk factor for CE subtype was heart disease.Conclusion The highest proportion of TOAST types in youth cerebral infarction group is small artery occlusion lacunar.Major risk factors for this group of youth cerebral infarction are as follows:hypertension,carotid atherosclerosis,heart disease,diabetes,hyperhomocys-teinemia,obesity,smoking and family history,and these risk factors should be actively intervened.
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Purpose To explore the value of spectral coronary CT angiography (CCTA) in reducing radiation dose and contrast dose using individualized optimal monochromatic imaging.Materials and Methods Sixty patients with suspected coronary disease were recruited in the study,who were randomly divided into two groups:group A (n=30) using conventional CT protocol with 350 mgI/ml contrast agent;group B (n=30) using low dose spectral CT imaging mode with 300 mgI/ml contrast agent.The images of group A were reconstructed with conventional process,and the images of group B were reconstructed with Optimal CNR to obtain the optimal monochromatic energy images.The images of both groups were transferred to an Advanced Workstation for analysis.Double-blinded method was carried out to qualify the images.CT values of coronary artery segments,as well as standard deviations (SD),the signal-to-noise ratio (SNR),contrast-to-noise ratio (CNR) of aortic sinus were measured.Radiation doses and iodine intake were compared between the two groups.The optimal keV distribution in group B was analyzed.Results There was no significant difference for the subjective scoring of image quality,CT value of each coronary artery segment,SD,SNR,and CNR values between the two groups (P＞0.05).The effective radiation dose and total iodine load in group B were less than that in group A (P＜0.05).The optimal energy distribution for group B was 60-75 keV,average at (66.50+3.91) keV.Conclusion Compared with the conventional CT protocol,spectral CT imaging at optimal energy levels combined with iterative reconstruction can effectively reduce the radiation dose and iodine load,and obtain better images than usual protocol.
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Objective To explore the application of low dose of radiation combined with low concentration of contrast medium in the energy spectrum CT of the coronary artery angiography.Methods 60 patients with suspected diagnosis of coronary heart disease were randomly divided into A,B two groups,30 cases in each group.Group A with 350 mg I/mL contrast agent,undergoing conventional CT scanning;Group B using 300 mg I/mL contrast agent,the gems energy spectrum CT scanning with low-dose.Two groups both adopted forward-looking heart switch control scanning mode.After scanning,group A reconstructed conventional images of 40% ASiR sequence,group B rebuilded axial surface images of single energy 65 keV and 40% ASiR sequence,all the reconstruction images were introduced to AW4.6 workstations used for analysis.Double-blind subjective rating was done by two experienced doctors to measure CT values and SD of aortic sinus (AS),left main (LMA),the left anterior descending branch proximal (LAD-p),left circumflex branch proximal (LCX-p), right coronary artery proximal (RCA-p)and pericardial fat,AS the noise (SD),to calculate the signal-to-noise ratio (SNR)and contrast to noise ratio (CNR),to calculate the effective doses of radiation(ED)by recording CTDI and DLP,and to record iodine intake.By using two independent samples t test to compare two groups of patients’the effective radiation doses,iodine intake,the average CT value,SD,SNR and CNR.Results The subjective image quality score differences and coronary measuring section CT values between the two groups had no statistical significance.In the aspects of effective radiation dose,group B reduced about 29% compared to group A,the difference was statistically significant.Iodine intake in group B decreased about 16% than in group A.Conclusion In coronary artery CT imaging, spectral scanning with low dose of radiation and reconstruction images with single energy can effectively reduce the radiation dose and iodine intake,at the same time it can obtain the approximate image quality like conventional scanning.
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Objective To evaluate the role of endoplasmic reticulum stress in ketamine-induced apoptosis in rat neurons.Methods Rat adrenal pheochromocytoma cell line (PC12 cells) was seeded in the culture dishes 100 mm in diameter (10 ml/dish) or in 6-well plates (2 ml/well) at a density of 5 × 105 cells/ml.PC12 cells were divided into 4 groups (n =6 each) using a random number table:control group (group C);ketamine group (group K);endoplasmic reticulum stress inhibitor salubrinal group (group S);ketamine + salubrinal group (group K+S).In group C,the cells were cultured in the plain culture medium.In group K,1.5 mmol/L ketamine was added.In group S,30 mmol/L salubrinal was added.In group K + S,1.5 mmol/L ketamine and 30 mmol/L salubrinal were added.At 24 h of incubation,the cell morphology was observed under light microscope,the expression of Bip and caspase-12 in PC12 cells was detected by Western blot,and the cell apoptosis was measured by flow cytometry.The apoptosis rate was calculated.Results Compared with group C,the expression of Bip and caspase-12 was significantly upregulated,and the apoptosis rate was increased in K and K + S groups (P ＜ 0.05),and no significant change was found in the parameters mentioned above in group S (P＞ 0.05).Compared with group K,the expression of Bip and caspase-12 was significantly down-regulated,and the apoptosis rate was decreased in group K+S (P＜0.05).The degree of damage to PC12 cells was more serious in group K than in group C..The degree of damage to PC12 cells in group K+S was significantly mnilder than that in group K and more serious than that in group C.Conclusion The mechanism by which ketamine induces neuronal apoptosis is related to the enhancement of endoplasmic reticulum stress in rats.
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PurposeTo explore the feasibility of personalized energy spectrum scanning in upper abdomen CT by comparing the image quality and radiation dose of optimizing choice spectrum scanning protocol with conventional 120 kVp scan.Material and Methods Sixty patients undergoing abdominal CT scan with and without contrast enhancement were prospectively collected and randomly assigned into two groups. Group A (30 patients) underwent conventional 120 kVp scan and spectral enhancement scanning; group B (30 patients) underwent spectrum scan and 120 kVp enhancement scanning. Spectral scanning protocol was based on individual choice with conventional 120 kVp NI10-5 mm average mAs scan for every patient. The CT dose index of volume (CTDIvol) and effective dose (ED) during non-contrast phase and portal venous phase were recorded. The CT value, standard deviation (SD), signal noise ratio (SNR) and contrast noise ratio (CNR) were measured in the liver parenchyma, spleen parenchyma and portal venous trunk.Results The CTDIvol and ED of spectrum scanning were less than 120 kVp scan but there was no statistical difference (P>0.05). The SD of group B GSI imaging was less than group A 120 kVp (P<0.05), while the SNR was greater than group A. The SD of group A GSI portal venous phase in the liver, spleen and portal vein was less than group B 120 kVp (P<0.05).ConclusionThe optimized energy spectrum scanning protocol can reduce radiation dose with quality of single energy image from the energy spectrum equal to or better than the conventional 120 kVp scanning protocol. Personalized energy spectrum scan protocol provides multi-parameter diagnosis and multi-application platform and can be used routinely in the upper abdomen scan.
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Objective To compare the virtual non-enhanced chest CT (VNCT)generated from spectral CT with conventional non-enhanced chest CT in patients with lung disease in terms of CT number accuracy and image quality.Methods A total of 30 patients with lung disease proved by pathology underwent the conventional non-enhanced thoracic CT and contrast enhanced CT with spectral imaging mode in arterial phase (AP)and venous phase (VP).The VNCT images were reconstructed based on the enhanced spectral CT imaging data.The mean CT number,signal to noise ratio (SNR)for the lesions and image quality score were obtained and compared between the true non-contrast CT (TNCT)and the VNCT (including AP and VP)with paired t test.Results The mean±standard deviation for CT number were (38.74±5.17)HU,(39.08±5.07)HU and (38.96± 5.18)HU for TNCT,VNCT at AP and VNCT at VP,respectively, with no statistical difference (P>0.05).All 3 sets of images demonstrated acceptable image quality,even though there were statistically significant differences in the SNR value and image quality score.The mean ± standard deviation values for SNR were 4.74±0.42 with TNCT, 3.79 ± 0.5 1 with VNCT at AP and 3.77 ± 0.39 with VNCT at VP (P <0.05),and the image quality scores were 5.00 ±0.00 with TNCT,4.17±0.65 with VNCT at AP and 4.17±0.53 with VNCT at VP (P<0.05).Conclusion In patients with lung disease,the vir-tual non-enhanced CT images generated from spectral CT provide accurate CT numbers for lesions and acceptable image quality com-pared with the true non-contrast CT.VNCT may be used to replace TNCT to improve work flow and reduce radiation dose.
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<p><b>OBJECTIVE</b>To investigate the relationship between fetal growth restriction and decreased ovarian reserve (DOR) in adulthood to screen high-risk population for early interventions.</p><p><b>METHODS</b>Forty-four patients with FGR and 88 normal women aged 18-40 years were enrolled. All the subjects were examined for serum levels of follicle-stimulating hormone (FSH), inhibin B (INH-B), and anti-mullerian hormone (AMH) using enzyme-linked immunosorbent assay method in the first 3-5 days of the menstrual cycle, and the counts of antrum ovarian follicle were detected by transvaginal or transabdominal ultrasonography.</p><p><b>RESULTS</b>The serum levels FSH, INHB, AMH, and AFC in FGR group differed significantly from those in the control group (P<0.05).</p><p><b>CONCLUSION</b>FGR will affect reproductive endocrine function in adulthood to cause a decreased ovarian reserve.</p>
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Adolescent , Adult , Female , Humans , Young Adult , Anti-Mullerian Hormone , Blood , Case-Control Studies , Enzyme-Linked Immunosorbent Assay , Fetal Growth Retardation , Follicle Stimulating Hormone , Blood , Inhibins , Blood , Menstrual Cycle , Ovarian Follicle , Physiology , Ovarian Reserve , Prospective StudiesABSTRACT
AIM:To observe the effects of cholecystokinin octapeptide (CCK-8) and its receptor antagonists on cAMP response element binding protein ( CREB) and phosphorylated CREB ( pCREB) expression in frontal cortex and hippocampus of morphine withdrawal rats , which aim to explore the post-receptor mechanism through which CCK-8 regu-lates morphine withdrawal .METHODS: After the morphine dependence and naloxone-precipitated withdrawal animal models were established, the effects of CCK-8, L-364718 (CCK1 receptor antagonist) and LY-288513 (CCK2 receptor an-tagonist) pretreatment on CREB and pCREB expression in frontal cortex and hippocampus were observed by Western blot -ting and immunohistochemistry .RESULTS:In rat frontal cortex neuron , CREB was expressed in both cytoplasm and nu-cleus, but pCREB was only highly expressed in the nucleus .In the pyramidal cell layer of hippocampal CA 1 region, CREB showed high expression in the cytoplasm and low expression in the nucleus , while pCREB was only expressed in the nu-cleus.No obvious change of CREB was observed after either chronic morphine treatment or naloxone withdrawal .The pCREB expression was increased after chronic morphine treatment and further increased after naloxone withdrawal .Com-pared with the withdrawal group , chronic pretreatment with CCK-8, L-364718 and LY-288513 had no effect on CREB expression in the frontal cortex , but obviously decreased the pCREB expression .In the hippocampus , pretreatment with L-364718 and LY-288513 decreased CREB and pCREB expression , but only the pCREB expression was decreased after CCK-8 treatment.CONCLUSION:CCK-8 and CCK receptor antagonists may alleviate morphine withdrawal symptoms by regulating CREB , with specificity in different brain regions .
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BACKGROUND:Establishing a characteristic.stable and repeatable model of Th1/Th2 imbalance in animals,is the key of studying the mechanism of Th1/Th2 imbalance.OBJECTIVE:To observe the characteristics of Th1/Th2 imbalance in splenocytes derived from Balb/C mice immnnized by keyhole limpet hemocyanin(KLH).DESIGN:A randomized control exploratory experiment.SETTING:Hebei Provincial Forensic Laboratory.Institute of Basic Medicine,Hebei Medical University.MATERLALS:The experiment was carried out in the Hebei Provincial Forensic Laboratory,Institute of Basic Medicine,Hebei Medical University from September 2005 to January 2007.Balb/C mice were adopted in this study.and all the disposals were in accordance with the guidance of animal ethics.METHODS:Balb/C mice were immunized with KLH emulsified in complete Freund's adjuvant(CFA),splenocytes were acquired,and the peak of cytokine secretion was determined in 3 groups:KLH groups of 6.25 mg,kg.12.5 mg,kg and 25 mg/kg.According to the immunizing dose and immunizing frequency.mice were divided into 7 groups:KLH groups of 6.25 mg/kg,12.5 mg/kg and 25 mg/kg,secondary immunity groups of 6.25 mg/kg,12.5 mg/kg and 25 mg/kg,as well as control group.According to the determined levels of IgG1 and IgG2a in blood serum.mice were divided into KLH group of 6.25 mg/kg and control group.MAIN OUTCOME MEASURES:Mice splenocytes supematant was detected with enzyme linked immunosorbent assay (ELISA)for the production of Th1 cytokines interferon (IFN)-γ,interleukin(IL)-2.IL-12 p40 and Th2 cytokines IL-4 and IL-5.The levels of Th1 antibody IgG2a and Th2 antibody IgG1 in blood serum were also detected by ELISA.RESULTS:The spleen derived from KLH-immunized mice appeared hypertrophy,and the number of splenocytes was manifold.Splenocytes restimulated with KLH in vitro produced much more IFN-γ,IL-2,IL-4,IL-5,but not IL-12p40.IL-2 secretion was obviously elevated after incubated for 24 hours and achieved pinnacle at 48 hours;productions of IL-4,IL-5 and IFN-γ were elevated after 24 hours,and increased gradually to 96 hours;IL-12p40 production was very low at every time point.Using different doses of KLH inlmunity once or twice,could all lead to the elevated productions of IL-2,IL-4,IL-5,IFN-γ,and the elevation of IL-4/IFN-γ ratio,but the secondary immunity group of 6.25 mg/kg KLH showed obviously higher levels than other groups(P＜0.01).The level of KLH specific antibody IgG2a and IgG1,especially IgG1 was elevated in blood serum of KLH-immunized mice.CONCLUSION:Balb/C mice immunizad with KLH emulsified in CFA can indce a Th2 predominant imbalance in splenocytes.
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BACKGROUND: MyoD gene is one of family members of muscle transcription factors. Transfection MyoD gene can switch on the procedure of differentiation of muscles, and transit non-muscle cells into muscle cells.The MyoD gene only expresses in skeletal muscles. Based on the same contractive structure in myocardial cells and skeletal muscle cells, it is imagined that the conversion from exogenous MyoD gene-induced fibroblast in local myocardium into skeletal muscle cells that had contractive function may become another method in the treatment of congestive heart failure on clinic.OBJECTIVE: To construct and identify a recombinant adenoviral vector carrying MyoD gene for further studies on the recovery function of MyoD gene in myocardial injury.DESIGN: Single sample experiment.SETTING: Department of Cardiology, First Affiliated Hospital, Nanjing Medical University.MATERIALS: The experiment was conducted at the Laboratory of Microbiology and Immunology, Nanjing Medical University between September 2004 and September 2005. MyoD gene and non-replicating form expressive vector of adenovirus were taken as research materials.METHODS: MyoD cDNA fragments were extracted from plasmids pEMSV-MyoD with polymerase chain reaction (PCR), and PCR was used to clone the whole-length gene of MyoD. After adding CACC sequence at 5' end, MyoD gene was cloned by orient topology into transfer ventor, pENTR/D-TOPO. Objective gene was transferred into adenoviral expression vector DNA via pENTR/D-TOPO vector. The recombinant adenoviral vectors transfected into HEK293A cells by using lipofectamine were packaged and amplified.MAIN OUTCOME MEASURES: Evaluation of PCR and DNA sequencing were used for confirming the size of segment and correctness of rank of MyoD cDNA and detecting the titre of virus.RESULTS: MyoD recombinant adenovirus contained target segment with precise length confirmed by PCR and DNA sequence that was correct. The titre of virus was 1.3×1011 pfu/mL.CONCLUSION: The recombinant adenoviral vector carrying MyoD gene is constructed successfully.
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0.05).But PMA increased and SC-3088 decreased cAMP content and PKA activity in LPS-stimulated rat PIMs(P
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Aim To investigate the effects and mechanisms of CCK-8 on IL-1? induced proliferation of RSC-364, a rat fibroblast-like synovial cell line. Methods MTT colorimetric assay and Western blot were used to measure cell proliferation and p38MAPK phosphorylation level to elucidate the mechanism of CCK-8 in IL-1? induced RSC-364 proliferation. Results CCK-8 significantly inhibited IL-1?-induced RSC-364 proliferation at 10 -12 , 10 -10 , 10 -8 , 10 -6 mol ? L -1 , and IL-1?-activated p38MAPK activity at 10 -10 , 10 -8 , 10 -6 mol?L -1 in a dose-dependent manner. The effect of CCK-8 was blocked by CR1409 (a CCKA-receptor antagonist) and CR2945 (a CCKB-receptor antagonist). Conclusion CCK-8 inhibits IL-1?-induced RSC-364 proliferation, probably by reducing p38MAPK activity through CCKA and CCKB receptors.
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Forensic toxicology is a specialized subject which forensic medical appraisers and students must learn and master. As a new teaching mode,problem based learning method is attrcting a lot of attention. Applying "case" based PBL teaching method is an important method to train innovative and high technical ability forensic talents.
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<p><b>OBJECTIVE</b>To study the effect of cholecystokinin octapeptide (CCK-8) on lipopolysaccharide (LPS)-stimulated pulmonary interstitial macrophages (PIM) in vitro.</p><p><b>METHODS</b>PIM were isolated and cultured in the presence or absence of LPS, CCK-8, proglumide (the antagonist of CCK receptors) and vehicle. The expression of membrane CD14 (mCD14) protein was assayed by flow cytometry and soluble CD14 (sCD14) in the supernatant was analyzed semi-quantitatively by Western blot. TNF-alpha in the supernatant was detected with ELISA.</p><p><b>RESULTS</b>CCK-8, at concentrations of 10(-7) mol/L and 10(-6) mol/L, significantly inhibited the expression of mCD14. Release of sCD14 and TNF-alpha in the supernatant was up-regulated by LPS (1 microg/ml) but reduced by CCK-8. The effect of CCK-8 was inhibited by proglumide.</p><p><b>CONCLUSION</b>CCK-8 negatively modulated several functions of LPS-stimulated PIM through CCK receptors. This may be one of the mechanisms for CCK-8 to alleviate inflammation in lung tissue during endotoxemia.</p>
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Animals , Female , Rats , Cells, Cultured , Culture Media, Conditioned , Chemistry , Lipopolysaccharide Receptors , Lipopolysaccharides , Pharmacology , Macrophages, Alveolar , Cell Biology , Metabolism , Rats, Sprague-Dawley , Sincalide , Pharmacology , Specific Pathogen-Free Organisms , Tumor Necrosis Factor-alpha , Bodily SecretionsABSTRACT
AIM: To investigate the inhibitory effects of cholecystokinin octapeptide(CCK-8) on nuclear factor-?B(NF-?B) activities stimulated by lipopolysaccharide(LPS) by using forskolin,the activator of adenylate cyclase,and PKA inhibitor H-89 in rat pulmonary interstitial macrophages(PIMs).METHODS: PIMs were isolated and purified.EMDA was applied to detect NF-?B activities and Western blotting was used to analyze the I?B-? protein level in rat PIMs.RESULTS: The NF-?B activity was not detected in normal control rat PIMs.The NF-?B activity in LPS-treated rat PIMs was obviously higher than that in control group(P0.05).The NF-?B activity in CCK+LPS group and LPS+Fsk group were obviously lower than that in LPS group(P
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AIM: To study the effect of cholecystokinin octapeptide(CCK-8) on lipopolysaccharide (LPS)-stimulated pulmonary interstitial macrophage(IM) in vitro. METHODS: Pulmonary IM were isolated and cultured in the presence of LPS, CCK-8, proglumide(the antagonist of CCK receptors) and vehicle alone or together. The expression of mCD14 protein was assayed by flow cytometry, and sCD14 in the supernatant was analyzed semi-quantitatively by Western blot, and TNF-? in the supernatant was detected with ELISA. RESULTS: CCK-8, at concentrations from 10 -7 mol/L to 10 -6 mol/L inhibited significantly the expression of mCD14, the release of sCD14 and TNF-? to the supernatant up-regulated by LPS(1 mg/L). The effect of CCK-8 was inhibited by proglumide. CONCLUSION: CCK-8 modulated negatively several functions of LPS-stimulated pulmonary IM through CCK receptors, which may be one of the mechanisms for CCK-8 to alleviate the inflammation in lung tissues during endotoxemia.