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1、第三军医大学博士学位论文常用英文缩略词表缩写英文全称中文译名ACAdenylate cyclase腺苷酸环化酶ALIAcute lung injury急性肺损伤ARDSAcute respiratory distress syndrome急性呼吸窘迫综合征AsODNAntisense oligodeoxynucleotide反义寡核苷酸DABDiaminobezidine二氨基联苯胺DEPCDiethyl pyrocarbonate焦碳酸二乙酯DTTDithiothreitol二硫苏糖醇 dNTPDeoxyribonucleic triphosphate脱氧核苷三磷酸 EDTAEthylene
2、 diaminetetraacetie acid乙二胺四乙酸 ETEndothelin内皮素IP3Inositol1，4，5-trisphosphatel，4，5-三磷酸肌醇LBILun：gbody weight index肺体指数mRNAMessenger RNA信使RNANFvBNuclear factorrJ3核转录因子K BP峨platelet activating factor血小板活化因子PLCPhospholipase C磷酯酶CPMSFPhenylmethylsulfonyl fluoride苯甲基磺酰氟RGSRegulator of G protein signalingG蛋
3、白信号转导调节因子RLWRate of lung water肺含水率TXA2Thromboxane A2血栓素A2第三军医大学博士学位论文The mechanism of Gq protein and RGS on ARDSAbstractAcute respiratory distress syndrome(ARDS)is one of the diseases that is severely harmful to humanS healthAlthough the death-rate of ARDS has been decreased in recent yearsit is sti
4、ll a difficult point in clinic therapyThe lung damage may be caused by directinjury of lung substance；or mass inflammatory mediators released after acute inflammatory reaction，which can cause exaggerated or uncontralled inflammatory responsenleExaggerated or uncontralled inflammatory response is one
5、 of the main reason of lunginjuryUptodate，themostdirectsignalof activationof exaggeratedoruncontralledinflammatory response iS still unclearBut it is shown that Gq proteincoupledreceptors(such as TXA2、PAF、ET receptors、play important roles in the process of acute lung injuryThese receptors have impor
6、tant role in platelet aggregationbronchospasm and endovascular microthrombosisEach of these Gq protein-coupled receptor antagonist couldpartly decrease lung edemaOf cause，many known or unknown cytokines also have rolesin ARDS pathogenesis，but which is the most important one is still hard to answer A
7、lthough cytokines and chemokines have a large number of members，signal pathways are fewer，and G protein signal pathway is one ofthe most important pathways which regulatenumerous cytokines and chemokines their effectsGq protein coupled receptors take part in the pathophysiology of acute lung injury，
8、SO regulation of Gq and its pathway maybe have significant role in this processsNewly discovered regulator ofG-protein signaling(RGS)is a GTP硒e activating protein(GAP)It can accelerate GTP hydrolyzing of Gia and Gqa，consequently reduce the intensity andduration of Gq and Gi protein activation，and ad
9、just the signal pathwayIn physiological conditionsRGS in organisms regulate G-protein pathway elaborately to make sure thatGprotein signal system Can response to the stimulating signal on some correct intensity anddurationIn pathological conditions，if RGS is out of control，their function of regulati
10、ng Gprotein pathway may be abnormal，thereby change the G protein signal transduction and do harm to tissueInthisstudy，we initially synthesized antisense oligonucleotide of Gq protein，and2第三军医大学博士学位论文observed its effect on inhibiting Gq protein expression in vitro and blocking the signal transduction
11、Next itS therapeutic role on acute lung injury in vivo was observed，and the role of Gq as well as the regulation mechanism of Gq on acute lung injury pathogenesis though the change of RGS was discussedMain contents and results：1ECV304 cell culture experiments showed that modified antisense oligonucl
12、eotide of Gqa by sulfur could inhibit Gqagene and protein expression effectivelyThis inhibitory effect are dosedependantFrom 01 ltM to 1 00 IxM，the antisense oligonucleotide inhibitory effect enganced with the increase of the dosage，and with the elongation of time on 24h， 48h and 72h pointsAntisense
13、 oligonucleotide of Gqu has no effect on Gs and Gi mRNAexpression2Detection of IP3 content using radioactive ligand binding experiment demonstrated that after stimulation with U46619，IP3 generated by ECV304 cells increased significantly (PO01)After treatment with antisense oligonucleotide of Gqa，the
14、 generation of IP3 inECV304 cells call be effectively inhibited in the basic condition and after U4661 9 treatmentBetween O1pM to 10，0 pM，this inhibition effect increased at dose-denpendent manlierThese results showed，manually synthesized antisense oligonucleotide could inhibit Gqu mediated signal t
15、ransduction though inhibition of Gqa3 Pretreatment of mouse with antisense oligonucleotide of Gqu Can ameliorate pulmonary edema and blood-gas ofpulmonary artery distressed by oleic acid in 6h and 24h After 6h and 24h of acute lung injury caused by oleic acid，expression of Gqu protein decreased significantly，but IP3 content in lung tissue increased markedly，it cued that Gqu activity reinforces during acute lung injuryPretreatment with
