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Expression and signifi cance of TLR4 and HIF-1α in pancreatic ductal adenocarcinoma

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World Journal of Gastroenterology 2010年第23期16卷 2881-2888页

作者： Jian-Jun Zhang,He-Shui Wu,Lin Wang,Yuan Tian,Jing-Hui Zhang,Hai-Long Wu department of Pancreatic Surgery,union hospital,tongji medical college,huazhong university of science and technology,wuhan 430022,Hubei Province,china department of Pediatrics,union hospital,tongji medical college,huazhong university of science and technology,wuhan 430022,Hubei Province,china Laboratory of General Surgery,union hospital,tongji medical college,huazhong university of science and technology,wuhan 430022,Hubei Province,china department of Pancreatic Surgery Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 Hubei Province China. heshuiwu@***

AIM:To investigate the expression of toll-like receptor(TLR) 4,nuclear factor-κB(NF-κB) p65 and hypoxiainducible transcription factor 1α(HIF-1α) in pancreatic ductal adenocarcinoma and their clinical ***:The mRNA of TLR4 and HIF-1α were investigated by real-time polymerase chain reaction in 30 cases of pancreatic ductal adenocarcinoma and its adjacent tissues,and expression of TLR4,NF-κB p65 and HIF-1α protein were detected by immunohistochemistry in 65 cases of pancreatic ductal adenocarcinoma tissues and 38 cases of corresponding adjacent *** relationship between TLR4 or HIF-1α and pathologic features,as well as the association between TLR4 and HIF-1α,were also ***-Meier method was used to assess the impact of expression of TLR4 and HIF-1α on survival of patients with pancreatic ***:The relative quantif ication of TLR4 and HIF-1α mRNA in tumor tissues was 0.81±0.10 and 0.87±0.11,respectively,signif icantly higher than that in adjacent tissues(0.81±0.10 vs 0.70±0.16,P=0.002;0.87±0.11 vs 0.68±0.13,P=0.000).The protein expression of TLR4,NF-κB p65 and HIF-1α in tumor tissues was 69.20%,66.15% and 70.80%,respectively,being signif icantly higher than that in adjacent normal tissues(69.20% vs 39.50%,P=0.003;66.15% vs 31.58%,P=0.001;70.80% vs 36.80%,P=0.001).There was no signif icant correlation between TLR4 or HIF-1α expression and the age,gender,tumor location,the degree of tumor differentiation in the patients(P>0.05).However,there was signif icant correlation between the expression of TLR4 or HIF-1α and tumor size,lymph node metastasis,venous invasion and clinical staging(P4 and HIF-1α had a signif icant impact on survival of patients with pancreatic ***:TLR4,NF-κB p65 and HIF-1α are overexpressed in pancreatic adenocarcinoma,TLR4 may be partly involved in up-regulating HIF-1α,and both synergestically promote development of pancreatic adenocarcinoma.

关键词： Pancreatic ductal adenocarcinoma Toll-like receptor 4 Nuclear factor-κB p65 Hypoxia-inducible factor 1

TLR4 signaling induced TLR2 expression in the process of mimic cerebral ischemia/reperfusion in vitro

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中国科学：生命科学英文版 2010年第2期53卷 223-228页

作者： SUN Peng1,ZHANG Qing2,HAN JiYuan1,TIAN Yuan3 & ZHANG JingHui3 1department of Emergency,union hospital,tongji medical college,huazhong university of science and technology,wuhan 430022.china 2department of Anesthesia,union hospital,tongji medical college,huazhong university of science and technology,wuhan 430022.china 3department of General Surgery,union hospital,tongji medical college,huazhong university of science and technology,wuhan 430022.china 1. department of Emergency Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 China2. Department of Anesthesia Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 China3. Department of General Surgery Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 China

Both TLR4 and TLR2 participated in the mediation of the inflammatory injury in the process of partial cerebral ischemia/***,it still remains unclear whether a crosstalk exists between TLR2 and TLR4 in ischemic cerebral *** the present study,we investigated the effect of TLR4 signaling on TLR2 expression during mimic cerebral I/R in ***-2 cells were cultured and treated with ischemia/reperfusion,then transfected with the plasmid pEGFP-H1/TLR4-siRNA,the plasmid pEGFP-H1/control sequence-siRNA and the blank plasmid,***,the expression of TLR2 and TLR4 mRNA and protein,NF-κB p65 mRNA and supernatant TNF-α level were significantly higher in ischemia/reperfusion treated cells than those lack of ischemia/reperfusion treatment,and as compared with those in ischemia/reperfusion treated cells without transfection,no significant differences about the above mentioned gene and protein expression were found in the blank plasmid tranfected cells and the plasmid pEGFP-H1/control sequence-siRNA transfected cells respectively,while the expression levels in the plasmid pEGFP-H1/TLR4-siRNA transfected cells were significantly ***,in order to determine the effects of pyrrolidinediethyldithiocarbamate (PDTC),an NF-κB inhibitor,on the TLR4-induced TLR2 expression in BV-2 cells treated with ischemia/reperfusion,it was found that TLR4 and TLR2 mRNA expressions in PDTC pretreated cells were significantly lower in comparison with normal saline pretreated cells and non-pretreated *** data suggested that TLR2 activation,signaled by TLR4 and regulated by NF-κB,might be directly involved play an important role in ischemia/reperfusion induced brain damage.

关键词： Toll-like receptor 4 Toll-like receptor 2 cerebral ischemia/reperfusion microglia

NMDA and AMPA receptors mediate intracellular calcium increase in rat cortical astrocytes

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Acta Pharmacologica Sinica 2004年第6期25卷 12-18页

作者： Bo HU, Sheng-gang SUN2, E-tang TONGNeurology department, union hospital, tongji college, huazhong university of science and technology, wuhan 430022, china Neurology department Union Hospital Tongji College Huazhong University of Science and Technology Wuhan 430022 China

AIM: To study the effect of glutamate on the intracellular calcium signal of pure cultured rat astrocytes and the roleof N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) re-ceptors in the procedure. METHODS: The fluorescence of calcium was measured by Fura-2/AM (F345/F380).RESULTS: L-Glutamate induced [Ca2+]i increase in most of the cells in concentration- and time-dependent *** 50 mmol/L induced the fluorescence increase by almost three to four times, while the effect of AMPA 50mmol/L was just half of that of D-(-)-2-amino-5-phosphonopentanoic acid (D-AP-5; a selective antagonist of theNMDA receptor). 6-Cyano-7-nitroquinoxaline-2,3-dione (CNQX, a selective antagonist of the AMPA receptor)abolished the effects of NMDA and AMPA, respectively. D-AP-5 and CNQX simultaneously or respectively attenu-ated the effect of L-glutamate at different degrees, but could not abolish it entirely. CONCLUSION: Glutamatemodulated intracellular Ca2+ of pure cultured rat astrocytes through different pathways. The activation of NMDAand AMPA receptors took part in the complex mechanisms.

关键词： glutamate astrocytes NMDA receptors AMPA receptors calcium

Hyaluronic acid fragments evoke Kupffer cells via TLR4 signaling pathway

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science china(Life sciences) 2009年第2期52卷 147-154页

作者： ZHANG JinXiang1, WANG Hui2, XIAO Qing3, LIANG HuiFang4, LI ZhuoYa4, JIANG ChunFang1, WU HeShui5 & ZHENG QiChang5 1 department of Emergency Surgery, union hospital Affiliated to huazhong university of science and technology, wuhan 430022, china 2 department of medical Genetics, tongji medical college Affiliated to huazhong university of science and technology, wuhan 430030, china 3 department of Ophthalmology, union hospital Affiliated to huazhong university of science and technology, wuhan 430022, china 4 department of medical Immunology, tongji medical college Affiliated to huazhong university of science and technology, wuhan 430030, china 5 department of General Surgery, union hospital Affiliated to huazhong university of science and technology, wuhan 430022, china 1. department of Emergency Surgery Union Hospital Affiliated to Huazhong University of Science and Technology Wuhan 430022 China 2. department of medical Genetics Tongji Medical College Affiliated to Huazhong University of Science and Technology Wuhan 430030 China 3. department of Ophthalmology Union Hospital Affiliated to Huazhong University of Science and Technology Wuhan 430022 China 4. department of medical Immunology Tongji Medical College Affiliated to Huazhong University of Science and Technology Wuhan 430030 China 5. department of General Surgery Union Hospital Affiliated to Huazhong University of Science and Technology Wuhan 430022 China

Kupffer cells, expressing toll-like receptor 4 (TLR4), play a central role in hepatic ischemia/reperfusion (I/R) injury. Hyaluronic acid (HA) fragments, degradative products of high-molecular-weight HA (HMW-HA), acquire the ability to activate immune cells under inflammatory conditions. Here we inves- tigated whether HA fragments could activate Kupffer cells and analyzed the underlying mechanism. Kupffer cells were isolated from wild-type mice (WT, C3H/HeN) and TLR4 mutant mice (C3H/HeJ) and HA fragments were produced by the methods of enzyme digestion and chromatography. Then Kupffer cells were stimulated by HA fragments or other control stimuli. The activation of Kupffer cells was estimated as the release of pro-inflammatory cytokines. The activation of p38 MAPK pathway of Kupffer cells was checked and blocking experiments were done as well. The results indicated that HA fragments acquired the ability to activate Kupffer cells in vitro, which was TLR4 dependent and not due to contamination of lipopolysaccharide. Experiments of p38 MAPK kinase inhibition by SB-203580 verified p38 MAPK was required in HA fragments induced Kupffer cells activation. This suggests that HA fragments, degradative products of one of the major glycosaminoglycans of the extracellular matrix, play critical roles in Kupffer cell activation mediated by TLR4 signaling pathway, which is, at least partially, de- pendent on p38 MAPK activation.

关键词： Kupffer cells hyaluronic acid fragments toll-like receptor 4 p38 MAPK lipoplysaccharide reperfusion injury liver

Antioxidative effect of propofol during cardiopulmonary bypass in adults

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Acta Pharmacologica Sinica 2004年第3期25卷 80-86页

作者： Shi-hai ZHANG2,Shou-yong WANG,Shang-long YAO department of anesthesiology,union hospital,tongji medicalcollege,huazhong universityof science & technology,wuhan 430022.china department of anesthesiology Union Hospital Tongji Medical College Huazhong University of Science & Technology Wuhan 430022 China

AIM: To investigate the antioxidative potential of propofol (an intravenous anesthetic with a chemical structure similarto phenol-basedfree radicalscavengers suchas vitaminE) during cardiopulmonary bypass(CPB). METHODS: Thirty adult patients referred for elective cardiac procedure with CPB were included and randomly allocated to a propofol group or a control group. Patients in the propofol group received propofol (0.1 mg·kg-1·min-1) intrave- nously for anesthesia maintenance, whereas those allocated to the control group received fentanyl 10 μg/kg intrave- nously and inhaled enflurane (1 %-1.5 %). Blood samples were collected at 7 time points: before the start of CPB, at 30 and 60 min of CPB, at the conclusion of CPB, 10 min after the administration of protamine, and 12 and 24 h after the cessation of CPB. Plasma levels of free F2-isoprostanes (sensitive markers of free radicals production) and complement C5a were determined by mass-spectrometricassay and enzyme immunoassay, respectively. Neu- trophil adhesion to endothelial cells was observed at ×200 magnificationunder a light microscope. RESULTS: Levels of F2-isoprostanes, complement C5a and neutrophil adhesion rate increased significantly during and after CPB in both groups. There were significantly higher levels of F2-isoprostanes, C5a, and more neutrophils adhering to endothelial cells in the control group than those in the propofol group, respectively. CONCLUSION: Cardio- pulmonary bypass is associated with a great production of damaging free radicals. Propofol may be beneficial both as an anesthetic and as a potent free radical scavenger in patients presenting pathologies associated with free radical reactions during CPB.

关键词： propofol cardiopulmonary bypass free radicals F2-isoprostanes complement 5a

Growth-inhibitory Effects of Curcumin on Ovary Cancer Cells and Its Mechanisms

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Journal of huazhong university of science and technology(medical sciences) 2004年第1期24卷 55-58页

作者：郑丽端童强松吴翠环 department of Pathology department of Pediatric Surgery

To study the growth inhibitory effects of curcumin on human ovary cancer A2780 cells in vitro and its molecular mechanisms, the growth inhibition rates of A2780 cancer cells, after being treated with 10 μmol/L-50 μmol/L curcumin for 6-24 h, were examined by MTT method. The morphological changes of cancer cells were observed under inversion microscopy. Cellular apoptotic rates were determined by using TUNEL. The protein expression levels of bcl 2, p53 and MDM2 in cancer cells were examined by SP immunohistochemistry. After being treated by various concentrations of curcumin, the growth of cancer cells was inhibited significantly. Some cancer cells presented characteristic morphological changes of apoptosis. The rates of apoptosis were 6.41 % -28.48 % ( P 3 was decreased, which depended on the action time ( P 3 was probably one of its molecular mechanisms.

关键词： curcumin ovary cancer gene expression apoptosis

Intercellular Adhension Molecule-1 in the Pathogenesis of Heroin-induced Acute Lung Injury in Rats

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Journal of huazhong university of science and technology(medical sciences) 2004年第5期24卷 430-432页

作者：周琼白明邹世清 department of Respiratory Diseases Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 China department of Withdrawal Liyuan Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430072 China

The expression of intercellular adhesion molecule-1 (ICAM-1) in the pathogenesis of heroin-induced acute lung injury (ALI) in rats was investigated. The model of ALI was established by intravenous injection of heroin into tail vein in rats. Thirty-six rats were randomly divided into heroin-treated groups (1 h, 2 h, 4 h, 6 h and 24 h) and normal control group. Changes in histopathologic morphology and biological markers of ALI were measured. The expression of ICAM-1 in lung tissue was detected by using immunohistochemistry and RT-PCR. The results showed that the W/D ratio and protein contents in BALF of the heroin-treated groups were significantly higher than that of the control group (P<0.01). The histopathological changes in the lung tissue were more obvious in heroin-treated groups. The ICAM-1 protein and mRNA expression in the lung tissue of heroin-treated groups were significantly increased as compared with that of the control group (P<0.01), and correlated with the ALI parameters in a time-dependent manner. Increasing of ICAM-1 expression was involved in the formation of heroin-induced lung injury. Furthermore, the level of expression was positively correlated with the severity of lung injury.

关键词： heroin acute lung injury intercellular adhesion molecule-1

Expression of CC Chemokine Ligand 20 and CC Chemokine Receptor 6 mRNA in Patients with Psoriasis Vulgaris

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Journal of huazhong university of science and technology(medical sciences) 2004年第3期24卷 297-299页

作者：吴艳李家文 department of Dermatology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 China

Summary: In order to explore the possible role of CC chemokine ligand 20 (CCL20) and its receptor CC chemokine receptor 6 (CCR6) in the pathogenesis of psoriasis, the expression levels of mRNA of them in psoriatic lesions were investigated. The skin biopsies were collected from skin lesions in 35 cases of psoriasis vulgaris and 18 normal controls. RT-PCR was used to semi-quantitatively analyze the mRNA expression of CCL20 and CCR6 in the psoriatic lesions and the normal skin tissues. The results showed that the mRNA of CCL20 and CCR6 was present in every specimen. The expression levels of CCL20 mRNA in skin lesions were 1.1397±0.0521, which were greatly higher than those in normal controls (0.8681±0.0308) (P3±0.0538, significantly higher than in the controls (0.9131±0.0433, P<0.001). These findings indicate that up-regulated expression of CCL20 and CCR6 mRNA might be related to the pathogenesis of psoriasis.

关键词： psoriasis CC chemokine ligand 20 CC chemokine receptor 6

The Anti-Proliferative Effect of Inhibitor of Telomerase on Cultured Retinal Pigment Epithelial Cells

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Journal of huazhong university of science and technology(medical sciences) 2001年第2期21卷 174-176页

作者：项奕曾水清 department of Ophthalmology Xiehe Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022

In order to provide a new method for treating proliferative vitreoretinopathy , the effects of anti proliferation and apoptosis induction of inhibitors of telomerase and heat shock protein 90 on the cultured retinal pigment epithelial cells were investigated. The rate of apoptosis cells was measured by using TUNEL on the cultured RPE cells, the co cultured RPE cells with inhibitor of telomerase or the co cultured RPE cells with inhibitor of Hsp90 . The cell proliferation status was measured in the above three groups by using MTT method. The rate of apoptosis in the RPE cells co cultured with camptothecin or geldanamycin was increased remarkably . MTT showed the rate of growth inhibition was 8.4 %, 32.3 % and 72.3 % at the concentrations of camptothecin 1 μmol/L, 5 μmol/L, 10 μmol/L, respectively, and 6.5 %, 30.9 %, 71.9 % at the concentrations of geldanamycin 1 μmol/L, 5 μmol/L, 10 μmol/L, respectively. It was concluded that telomerase and Hsp90 can promote the proliferation of the cultured RPE cells, while the inhibitor of them can induce apoptosis and inhibit the growth of the RPE cells.

关键词： camptothelin geldanamycin anti proliferative apoptosis