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Effect of nitric oxide on toll-like receptor 2 and 4 gene expression in rats with acute lung injury complicated by acute hemorrhage necrotizing pancreatitis

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Hepatobiliary & Pancreatic Diseases International 2005年第4期4卷 609-613页

作者： He-Shui Wu, Lei Zhang, Yan Chen, Xing-Jun Guo, Lin Wang, Jian-Bo Xu, Chun-You Wang and Jing-Hui Zhang Center of Pancreatic Surgery Laboratory of General Surgery affiliated Union hospital of tongji medical college, Hua-zhong university of science and technology, wuhan 430022, china.pe-diatric department, affiliated tongji hospital of tongji medical college, huazhong university of science and technology, wuhan 430030, china. Center of Pancreatic Surgery Affiliated Union Hospital of Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 China.

BACKGROUND: Toll-like receptor (TLR) 2/4 might play important roles in mediating proinflammatory cytokine synthesis and release. And nitric oxide (NO) has been used to treat acute respiratory distress syndrome (ARDS). This study aimed to investigate the changes in TLR2/4 gene expression in the lungs of rats with acute lung injury (ALI) complicated by acute hemorrhage necrotizing pancreatitis (AHNP) and the effect of NO on the TLR2/4 gene expression. METHODS: One hundred and ten SD male rats were randomly divided into sham-operated group ( n = 10) , AHNP group (n = 30) , chloroquine-treated group ( n = 30) , and L-Arg-treated group (n =40). The lungs were dissected for lung histological scoring, and bronchoalveolar lavages were harvested for lung injury indexing. TLR2/4 mRNA expression in the lungs was measured by RT-PCR. RESULTS: TLR2/4mRNA was detected in the lungs with low values in the sham-operated group (0.016±0. 210E-2, 0.112 ±0.750E-2) , but it was markedly increased at 3 hours in the AHNP group (0.787±0.751E-2, 1.512 ±1.794E-2) , peaking at 12 hours (1.113 ±6.141E-2, 2.957±2.620E-2; P <0.05 or P <0.01). When lung injuries were aggravated, TNF-α concentrations in the lungs were increased, but NO concentrations were decreased ( P < 0.05 or P < 0.01 ) . When TLR2/4mRNA was inhibited by CQ (3h: 0.313 ± 5.491E-2, 0.005 ±1.419E-3 ; 6h: 0.488 ±7.442E-2, 0.010 ± 1.518E-3; 12h: 0.883 ± 8.911E-2, 0.024 ± 2.760E-3; P< 0.05 or P <0.01) , lung injuries were relieved. NO concentrations in the lungs were increased but TNF-α concentrations were decreased (P <0. 05 or P <0.01). When the rats with AHNP were treated with L-Arg, TLR2/4mRNA expression in the lungs could be effectively inhibited (50mg-T: 0.656 ±3. 977E-2, 1. 501 ±6.111E-2; 100mg-T: 0.260± 0.891E-2, 0.732 ±5.135E-2; 200mg-T: 0.126 ±0.914E-2, 0.414 ± 1.678E-2; 400mg-T: 0.091 ±0.399E-2, 0.287 ± 0.176E-2; P <0.05 or P <0. 01) and lung injuries were relieved. At the same time, NO concentrations in the lungs were marke

关键词： toll-like receptors acute hemorrhage necrotizing pancreatitis lung nitric oxide chloroquine

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Acta Pharmacologica Sinica 2006年第9期27卷 1159-1164页

作者： Jin QIN Zheng-xiang LIU~1 department of Cardiology,tongji hospital,tongji medical college,huazhong university of science and technology,wuhan 430030,china department of Cardiology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China

Aim:To examine the inhibitory effect of FAK-related nonkinase(FRNK)in cardiachypertrophy in vitro and investigate the possible ***:A func-tional fragment of FRNK cDNA was amplified by reverse transcription-polymerasechain reaction and cloned into the vector *** in neonatal ratcardiac myocytes was established with angiotensin-Ⅱ *** pcDNA3.1-FRNK or pcDNA3.1 was respectively transfected into cardiomyocytes byLipofectamine *** surface area and mRNA expression of atrial natriureticpeptide(ANP)of myocytes were employed to detect cardiac ***-κBp65 protein in nuclear extracts,phosphorylation levels of ERK1/2(p-ERK 1/2)andAKT(p-AKT),as well as total ERK1/2,and AKT in variant treated cardiomyocyteswere determined by Western ***:Under the stimulation of angiotensin Ⅱ,the surface area of myocytes and levels ofANP mRNA were significantly *** transient transfection with pcDNA3.1-FRNK in advance may reduce the sur-face area and expression of ANP mRNA of hypertrophic *** proteinlevels of NF-κB p65 in nuclear extracts and p-ERKI/2,p-AKT in FRNK treatedcardiomyocytes were significantly decreased compared with that in angiotensin-Ⅱ induced cardiomyocytes,while different treatments had little effect on totalERK1/2 and ***:FRNK may inhibit angiotensin-Ⅱ-inducedcardiomyocyte hypertrophy via decreasing phosphorylation levels at ERK1/2 andAKT,consequently downregulating nuclear translocation of NF-κB p65.

关键词： FAK-related nonkinase angiotensin Ⅱ cardiomyocyte hypertrophy extracellular signal-regulated kinase 1/2 protein kinase B NF-kappaB p65

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CD151 gene delivery increases eNOS activity and induces ECV304 migration, proliferation and tube formation

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Acta Pharmacologica Sinica 2007年第1期28卷 66-72页

作者： Zhen-zhong ZHENG Zheng-xiang LIU~2 department of Cardiology,tongji hospital,tongji medical college,huazhong university of science and technology,wuhan 430030,china department of Cardiology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China

Aim:To investigate the effects of CD 151 on the activity of endothelial NO syn-thase(eNOS),and ECV304 migration,proliferation and tube ***:pAAV-CD 151 and pAAV-anti-CD 151 were constructed and used to transientlytransfect ECV304 mediated with Lipofectamine *** transfection,the ex-pression of CD 151 was measured by Western *** migration assay wasperformed using Boyden transwell;proliferation assay was evaluated using the 3-[4,5-dimethylthiazol-2-yl]-2,5,diphenyltetrazolium bromide(MTT)method,andtube formation test was examined on matrigel,eNOS activity was assayed by L-[~3H]citrulline production from L-[~3H]*** involvement of eNOS was ex-plored using an eNOS inhibitor(L-NAME)and the effects in the process ***:CD 151 promotes cell migration,proliferation and tube *** addition,CD151 increases eNOS ***,cell migration,prolifera-tion and tube formation induced by CD 151 are inhibited when L-NAME is used,which indicates that there is an involvement of eNOS in CD151-induced cellmigration,cell proliferation and tube ***:CD 151 promotesECV304 migration,proliferation and tube *** mechanism is that CD151increases eNOS *** result also suggests that eNOS is involved in theangiogenic effects of CD151.

关键词： CDI51 endothelial NO synthase(eNOS) cell migration cell proliferation tube formation

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Deguelin regulates nuclear pore complex proteins Nup98 and Nup88 in U937 cells in vitro1

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Acta Pharmacologica Sinica 2005年第10期26卷 1265-1273页

作者： Hong-li LIU~2, Yan CHEN~(2,3), Guo-hui CUI~2, Qiu-ling WU~2, Jing HE~2, Wei-hua CHEN~2, Jian-feng ZHOU~4 ~2 department of Hematology, Union hospital, tongji medical college, huazhong university of science and technology, wuhan 430022 ~4 department of Hematology, tongji hospital, tongji medical college, huazhong university of science and technology, wuhan 430030, china department of Hematology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China department of Hematology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China

Aim: To investigate the anticancer effects and the molecular mechanisms of deguelin on human U937 leukemia cells, and to explore the underlying mechanism regulating nucleoporin 98 （Nup98） and nucleoporin 88 （Nup88） in vitro. Methods: The effects of deguelin on the growth of U937 cells were studied by MTT assay. The effect of deguelin on the cell cycle of U937 cells was studied by using a propidium iodide method. The localization of the nuclear pore complex proteins Nup98 and Nup88 was investigated by using immunofluorescence and immunoelectron microscopy. The expression of Nup98 and Nup88 in U937 cells was investigated by using flow cytometry and Western blot. Results: The prolif eration of U937 cells was inhibited in the deguelin-treated group, with a 24-h IC50value of 21.61 nmol/L and a 36-h IC50value of 17.07 nmol/L. U937 cells treated with deguelin had reduced percentages of cells in the G0/G1phase, whereas cells accumulated in the S and G2/M phases. Nup88 and Nup98 were found on both the nuclar and cytoplasmic sides of the U937 cells by using immunofluorescence and immunoelectron microscopy. The expression of Nup98 was upregulated and that of the Nup88 protein was downregulated in U937 cells treated with deguelin. Conclusion: Deguelin is able to inhibit the proliferation of U937 cells by regulating the cell cycle such that cells are arrested at the S and G2/M phases, so that the proportion of cells in the G0/G1phase decreases. The antitumor effects of deguelin are related to upregulating the expression of Nup98 and downregulating the expression of Nup88 protein in U937 cells.

关键词： deguelin U937 cells nucleoporin Nup98 Nup88

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Curcumin,a potent anti-tumor reagent,is a novel histone deacetylase inhibitor regulating B-NHL cell line Raji proliferation

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Acta Pharmacologica Sinica 2005年第5期26卷 603-609页

作者： Hong-li LIU~2,Yan CHEN~(2,3),Guo-hui CUI~2,Jian-feng ZHOU~4~2department of Hematology,Union hospital,tongji medical college,huazhong university of science and technology,wuhan 430022,china.~4department of Hematology.tongji hospital,tongji medical college,huazhong university of science and technology,wuhan430030,china department of Hematology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China department of Hematology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China

Aim:To investigate curcumin(diferuloylmethane)induced apoptosis and its mo-lecular mechanism of action in B-NHL cell line Raji ***:Raji cells werecultured in RPMI-1640 medium and treated with curcumin in different concentra-tions.3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium(MTT)assay wasused to detect growth inhibition and Hoechst 33258 staining was used to *** and Western blot were used to detect the ex-pressions of histone deacetylase 1,3,and 8(HDAC 1,HDAC3,and HDACS)andacetylated histone H4(Ac-histone H4)***:Curcumin inhibited theproliferation of B-NHL cell line Raji cells with a 36-h IC50value of 24.1±2.0μmol/*** 33258 staining showed that curcumin could induce Raji cell *** expression levels of HDAC1,HDAC3,and HDAC8 proteins were downregulat-ed following curcumin treatment in Raji cells,whereas Ac-histone H4 protein expres-sion was upregulated after treatment with ***:Curcumin,as anew member of the histone deacetylase inhibitors,can inhibit the expression ofclass I HDACs(HDAC1,HDAC3,and HDAC8),and can increase the expressionof Ac-histone H4 in Raji *** plays an important role in regulatingB-NHL cell line Raji cell proliferation and apoptosis.

关键词： curcumin histone deacetylase acetylated histone H4 Raji

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Expression of Androgen Receptor in Meningiomas

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Journal of huazhong university of science and technology(medical sciences) 2001年第2期21卷 140-142页

作者：陈坚陈刚 department of Neurosurgery Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan

In order to investigate the expression of androgen receptor in meningiomas and its relation to tumor proliferative potential, we examined the expression of AR and proliferating cell nuclear antigen by avidine biotin complex immunohistochemistry in 39 cases of meningiomas. Of the 39 cases of meningiomas, 20 showed positive AR immunoreactivity. The AR expression positivity rates were 31 % in benign meningiomas, 58 % in atypical meningiomas, 87.5 % in malignant meningiomas, respectively. In addition to the tumor cells, cells of microvascular endothelial proliferation were frequently AR positive. Malignant meningiomas had a significantly higher percentage of AR positive cells compared with atypical and benign meningiomas . The mean proliferating cell nuclear antigen labeling index was significantly higher in the malignant meningiomas when compared with atypical meningiomas and benign meningiomas . AR positive meningiomas had higher PCNA LI than AR negative meningiomas . The expression of AR in tumor tissues was significantly related with PCNA LI. These data indicated that AR in the meningiomas was correlated with histological grade and AR might participate in the growth of these tumors and tumor angiogenesis. The measurement of AR in these tumors may indirectly represent tumor growth potential.

关键词： meningiomas androgen receptor immunohistochemistry proliferating cell nuclear antigen angiogenesis

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Passive sensitization increases histamine-stimulated calcium signaling and NF-κB transcription activity in bronchial epithelial cells

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Acta Pharmacologica Sinica 2006年第6期27卷 708-714页

作者： Si JIN~(2,5)Dan TIAN~3 Jian-guo CHEN~4 Li-ping ZHU~2 Sheng-yuan LIU~2 Di-xun WANG~2 2 department of Pathophysiology,tongji medical college,huazhong university of science and technology Key Laboratory of Pulmonary Disease of Ministry of Health of china.wuhan 430030,china.3 department of Pathology,tongji medical college.huazhong university of science and technology.wuhan 430030,china.4 department of Pharmacology,tongji medical college,huazhong university of science and technology,wuhan 430030,china department of Pathophysiology Tongji Medical College Huazhong University of Science and Technology Key Laboratory of Pulmonary Disease of Ministry of Health of china Wuhan China department of Pathology Tongji Medical College Huazhong University of Science and Technology Wuhan China department of Pharmacology Tongji Medical College Huazhong University of Science and Technology Wuhan China

Aim:To find out if the two aspects of asthma（chronic airway inflammation andbronchial hyperresponsiveness）are related to hypersensitivity of calcium signal-ing in bronchial epithelial ***:Porcine bronchial epithelial cells（PBEC）were divided into sensitized（S）and nonsesitized（N）*** group S,the cellswere preincubated with serum from ovalbumin sensitized guinea *** group N,the cells were preincubated with serum from nonsensitized guinea *** calcium imaging and ELISA-based NF-κ3 activity were used to evaluate thehistamine-stimulated intracellular free calcium level and NF-κB activity,***:First,0.1μmol/L histamine could induce [Ca2+]ioscillationsin PBEC of group S,but not in group ***,1/Jmol/L histamine could induce[Ca2+]ioscillations of PBEC in both group S and group *** [Ca2+]ioscillationfrequency of PBEC was significantly higher in group S than in group N,thoughthe[Ca2+]ioscillation amplitude showed no difference between the two ***,when 10 μmol/L histamine was used to stimulate PBEC a transient initialincrease followed by a sustained elevation（FSE）of [Ca2+]iwas observed in PBECin both *** amplitude of the FSE of [Ca2+]iin PBEC was significantlyhigher in group S than in group *** subsequent NF-κB activity was in accor-dance to the calcium oscillation frequency evoked by histamine,but not to ***:It was suggested that the increased sensitivity of calciumsignaling in bronchial epithelial cells might contribute to the exorbitant inflamma-tion or increased susceptibility in asthmatic airway epithelial cells.

关键词： asthma histamine calcium signaling bronchi epithelial cell nuclear factor of kappaB

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Influences of dibutyryl cyclic adenosine monophosphate and forskolin on human sperm motility in vitro

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Asian Journal of Andrology 2003年第2期5卷 113-115页

作者： Ji-Hong LIU, Yang LI, Zheng-Guo CAO, Zhang-Qun YEdepartment of Urology, tongji hospital, tongji medical college, huazhong university of science and technology,wuhan 430030, china department of Urology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China

Aim: To study the influences of dibutyryl cyclic adenosine monophosphate (dbcAMP) and forskolin on human sperm motility in vitro. Methods: Semen samples, aseptically obtained by masturbation and prepared by swim-up technique from 20 fertile men, were incubated with different concentrations of dbcAMP and forskolin at 37 癈. Measurements were carried out after 10 min, 20 min, 30 min and 60 min incubation. Motility parameters were estimated by using an automatic analyzing system. Results: Treatment with dbcAMP or forskolin resulted in a significant increase in sperm motility and progressive motility. The larger the concentrations of dbcAMP or forskolin, the greater the effect appeared. The straight linear velocity and curvilinear velocity were not affected by both agents. Conclusion: dbcAMP and forskolin increase the motility and progressive motility of human sperm in vitro.

关键词： dibutyryl cyclic AMP forskolin sperm motility

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Effect of telmisartan on expression of protein kinase C-α in kidneys of diabetic mice

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Acta Pharmacologica Sinica 2007年第6期28卷 829-838页

作者： Li-jun YAO~(2,4),Jian-qing WANG~2,Hong ZHAO~3,Jian-she LIU~2,An-guo DENG~2~2 department of Nephrology,Union hospital,tongji medical college,huazhong university of science and technology wuhan 430030,china.~3 department of Trauma Surgery,tongji hospital,tongji medical college,huazhong university of science and technology,430030wuhan.china department of Nephrology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China department of Trauma Surgery Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China

Aim:To investigate the effects of angiotensin receptor blocker (ARB) telmisartanon the expression and distribution of protein kinase C (PKC)-α in the kidneys ofdiabetic ***:Diabetic mice were induced with streptozotocin and agroup of them were randomly selected for treatment with *** 6weeks,the expression and localization of PKC-α in the renal cortex,and the outerand inner medulla were assessed by immunohistochemistry and semiquantitativeWestern *** addition,expressions of PKC-α,transforming growth factor-β1 (TGF-β1),and vascular endothelial growth factor (VEGF) in glomeruli weremeasured by semiquantitative ***:Diabetic and nor-mal mice showed similar distributions of PKC-α in the *** expression ofPKC-α was found in glomeruli,epithelial cells of proximal tubules,and medullary-collecting duct,while not in the medullary and cortical thick ascending limb,andwas different in the epithelial cells of proximal tubules of diabetic nephropathy(DN) mice,PKC-α was mostly translocated from the basement membrane to theapical membrane,whereas it was largely translocated from the apical membrane tothe basement membrane in epithelial cells of the inner medullary-collecting *** blotting detected increased expression of PKC-α in the renal cortex andouter medulla,but not in the inner medulla of DN *** expressions ofPKC-α,TGF-β1,and VEGF were shown in the glomeruli of DN mice,where PKC-αexhibited a correlation to VEGF,but no correlation to TGF-β*** telmisartanattenuated alterations of PKC-α as mentioned earlier in the DN ***:Our findings suggest that PKC-α may play a role in the pathogenesis of DN,andthat the nephroprotective effects of ARB telmisartan may be partly associatedwith its influence on PKC-α.

关键词： diabetic nephropathy protein kinases C-α telmisartan mouse kidney

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The Expression of Cyclins in Neurons of Rats after Focal Cerebral Ischemia

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Journal of huazhong university of science and technology(medical sciences) 2008年第1期28卷 60-64页

作者：陈彬王伟 department of Neurology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology

The change of the expression of Cyclins in neurons of rats after focal cerebral ischemia was investigated. Ischemia was induced by temporary middle cerebral artery occlusion （MCAO）. The experimental rats induced by MCAO were sacrificed on 7th and 14th day after reperfusion. The brain was taken out at 7th and 14th day after injury, and the expression of Cyclin D1, E, A and B1 in neurons of cerebral cortex or hippocampal CA1 region was detected by immunofluorescence and confocal microscope. The results showed that after MCAO, in the ipsilateral CA1 subfield of hippocampus the expression of Cyclin D1, E, A and B1 in neurons was significantly gradually up-regulated at 7th and 14th day after reperfusion （P〈0.05） as compared with that in control group. In the ipsilateral cerebral cortex the expression of Cyclin D1 and B1 in neurons was notably gradually down-regulated at 7th and 14th day, and that of Cyclin E and A was significantly up-regulated at 14th day after reperfusion as compared with that in control group （all P〈0.05）. It was concluded that there was a differential sensitivity among neurons from different brain regions to ischemic injury. But all of them re-enter into cell cycle after MCAO.

关键词： cerebral ischemia neuron cell cycle confocal microscope

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